Methods and compositions for occluding blood vessels, ablating tissue, and / or deliverying agents to localized areas within a mammal

Abstract

This document relates to methods and materials for treating cancer. For example, a composition provided herein including choline geranate, a sugar such as glycerol, and optionally one or more other compounds or agents can be used to ablate at least some cancer cells of a tumor (e.g., a solid tumor) within a mammal. In some cases, a composition provided herein including choline geranate, a sugar such as glycerol, and optionally one or more other compounds or agents can be administered to a mammal (e.g., a human) having cancer to ablate at least some cancer cells of a tumor (e.g., a solid tumor) within the mammal (e.g., to treat the mammal).

Description

[0001]METHODS AND COMPOSITIONS FOR OCCLUDING BLOOD VESSELS, ABLATING TISSUE, AND / OR DELIVERYING AGENTS TO LOCALIZED AREAS WITHIN A MAMMAL CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Patent Application Serial No. 63 / 602,949, filed on November 27, 2023. The disclosure of the prior application is considered part of (and is incorporated by reference in) the disclosure of this application. BACKGROUND Cancer is a leading cause of morbidity and mortality with an estimated annual rate of 10 million deaths worldwide (World Cancer Report 2014, (World Health Organization, 2014)) costing >$200 billion annually in the US alone (Mattiuzzi et al., J. Epidemiol. Glob. Health, 9:217-222 (2019)). Hepatocellular carcinoma (HCC) is the most common type of liver cancer. Once diagnosed, 5-year survival for regional or metastatic liver cancer is 9% and 3%, respectively. SUMMARY One aspect of the present disclosure related to a composition comprising an ionic liquid comprising geranic acid anions (or geranate anions) and choline cations, and a sugar, wherein the ratio of said geranate to said choline of said composition is from 6:1 to 1:6, optionally from 4:1 to 1:4, and further optionally from 3:1 to 1:3. In some embodiments, said composition comprises at least 30 percent of said ionic liquid. In some embodiments, said composition comprises at least 35 percent, of said ionic liquid. In some embodiments, said composition comprises at least 40 percent of said ionic liquid. In some embodiments, said composition comprises least 20 percent of said sugar. In some embodiments, said composition comprises at least 25 percent, of said sugar. In some embodiments, said composition comprises from 25 to 35 percent of said sugar. In some embodiments, the molar ratio of said geranic acid anions to said choline cations is from 1:1.5 to 1:6, optionally from 1:2 to 1:6. In some embodiments, the molar ratio of said geranic acid anions to said choline cations is from 1:2.5 to 1:6. In some embodiments, the molar ratio of said said geranic acid anions to said choline cations is from 1:2.5 to 1:5. In some embodiments, the molar ratio of said geranic acid anions to said choline cations is (a) from 1:3 to 1:6, (b) from 1:3 to 1:5, (c) from 1:3 to 1:4, (d) from 1:3.5 to 1:6, (e) from 1:3.5 to 1:5, (f) from 1:3.5 to 1:4, or (g) from 1:3.5 to 1:4.5. In some embodiments, the molar ratio of said geranic acid anions to said choline cations is 1:4. In some embodiments, said composition comprises one or more imaging agents. In some embodiments, said composition comprises (a) from about 1 percent (v / v) to about 40 percent (v / v) of at least one of said one or more imaging agents, (b) from about 0.01 mg to about 500 mg of at least one of said one or more imaging agents per mL of said composition, (c) from about 0.05 mg to about 250 mg of at least one of said one or more imaging agents per mL of said composition, (d) from about 0.1 mg to about 100 mg of at least one of said one or more imaging agents per mL of said composition, or (e) from about 0.1 mg to about 10 mg of at least one of said one or more imaging agents per mL of said composition. In some embodiments, (a) said composition comprises at least 1 percent of said one or more imaging agents, (b) said composition comprises at least 5 percent of said one or more imaging agents, (c) said composition comprises at least 10 percent of said one or more imaging agents, (d) said composition comprises at least 15 percent of said one or more imaging agents, (e) said composition comprises from about 1 percent to about 30 percent of said one or more imaging agents, (f) said composition comprises from about 5 percent to about 30 percent of said one or more imaging agents, (g) said composition comprises from about 10 percent to about 30 percent of said one or more imaging agents, or (h) said composition comprises from about 15 percent to about 25 percent of said one or more imaging agents. In some embodiments, said composition comprises two or more imaging agents. In some embodiments, said imaging agents are selected from the group consisting of iohexol, indocyanine green, and tantelum. In some embodiments, said composition comprises one or more therapeutic agents. In some embodiments, said composition comprises (a) from about 1 mg to about 50 mg of at least one of said one or more therapeutic agents per mL of said composition, (b) from about 0.01 mg to about 500 mg of at least one of said one or more therapeutic agents per mL of said composition, (c) from about 0.05 mg to about 250 mg of at least one of said one or more therapeutic agents per mL of said composition, (d) from about 0.1 mg to about 100 mg of at least one of said one or more therapeutic agents per mL of said composition, or (e) from about 0.1 mg to about 10 mg of at least one of said one or more therapeutic agents per mL of said composition. In some embodiments, (a) said composition comprises at least 1 percent of said one or more therapeutic agents, (b) said composition comprises at least 5 percent of said one or more therapeutic agents, (c) said composition comprises at least 10 percent of said one or more therapeutic agents, (d) said composition comprises at least 15 percent of said one or more therapeutic agents, (e) said composition comprises from about 1 percent to about 30 percent of said one or more therapeutic agents, (f) said composition comprises from about 5 percent to about 30 percent of said one or more therapeutic agents, (g) said composition comprises from about 10 percent to about 30 percent of said one or more therapeutic agents, or (h) said composition comprises from about 15 percent to about 25 percent of said one or more therapeutic agents. In some embodiments, said composition comprises two or more therapeutic agents. In some embodiments, a molecular weight of at least one of said one or more therapeutic agents is (a) greater than 7,500 Daltons, (b) greater than 10,000 Daltons, (c) greater than 15,000 Daltons, or (d) greater than 25,000 Daltons. In some embodiments, a molecular weight of at least one of said one or more therapeutic agents is (a) greater than 50,000 Daltons, (b) greater than 100,000 Daltons, (c) greater than 125,000 Daltons, or (d) greater than 150,000 Daltons. In some embodiments, a molecular weight of at least one of said one or more therapeutic agents is (a) less than 750,000 Daltons, (b) less than 500,000 Daltons, (c) less than 250,000 Daltons, or (d) less than 200,000 Daltons. In some embodiments, a molecular weight of at least one of said one or more therapeutic agents is (a) from 7,500 Daltons to 750,000 Daltons, (b) from 10,000 Daltons to 500,000 Daltons, (c) from 15,000 Daltons to 250,000 Daltons, or (d) from 20,000 Daltons to 200,000 Daltons. In some embodiments, at least one of said one or more therapeutic agents comprises a biologically active polypeptide. In some embodiments, said biologically active polypeptide is selected from the group consisting of nivolumab, ipilimumab, pembrolizumab, atezolizumab, durvalumab, cemiplimab, and avelumab. In some embodiments, at least one of said one or more therapeutic agents comprises an immunogenic polypeptide that is immunogenic within a mammal. In some embodiments, said mammal is a human. In some embodiments, at least one of said one or more therapeutic agents comprises an antigen-binding polypeptide. In some embodiments, said antigen-binding polypeptide is an antibody. In some embodiments, said antigen-binding polypeptide is selected from the group consisting of nivolumab, ipilimumab, pembrolizumab, atezolizumab, durvalumab, cemiplimab, and avelumab. In some embodiments, at least one of said one or more therapeutic agents has anti-cancer activity within a mammal. In some embodiments, said mammal is a human. In some embodiments, said one or more therapeutic agents having anti-cancer activity are selected from the group consisting of doxorubicin and cisplatin. In some embodiments, said composition comprises a nucleic acid. In some embodiments, said composition comprises (a) from about 5 mg to about 100 mg of said nucleic acid per mL of said composition, (b) from about 0.01 mg to about 500 mg of said nucleic acid per mL of said composition, (c) from about 0.05 mg to about 250 mg of said nucleic acid per mL of said composition, (d) from about 0.1 mg to about 100 mg of said nucleic acid per mL of said composition, or (e) from about 0.1 mg to about 10 mg of said nucleic acid per mL of said composition. In some embodiments, (a) said composition comprises at least 1 percent of said nucleic acid, (b) said composition comprises at least 5 percent of said nucleic acid, (c) said composition comprises at least 10 percent of said nucleic acid, (d) said composition comprises at least 15 percent of said nucleic acid, (e) said composition comprises from about 1 percent to about 30 percent of said nucleic acid, (f) said composition comprises from about 5 percent to about 30 percent of said nucleic acid, (g) said composition comprises from about 10 percent to about 30 percent of said nucleic acid, or (h) said composition comprises from about 15 percent to about 25 percent of said nucleic acid. In some embodiments, said nucleic acid encodes an imaging agent. In some embodiments, said imaging agent is selected from the group consisting of blue fluorescent protein (BFP), green fluorescent protein (GFP), and red fluorescent protein (RFP). In some embodiments, said nucleic acid encodes a therapeutic agent. In some embodiments, said therapeutic agent encoded by said nucleic acid is selected from the group consisting of a biologically active polypeptide, an immunogenic polypeptide, and an antigen-binding polypeptide. In some embodiments, said therapeutic agent encoded by said nucleic acid has anti-cancer activity within a mammal. In some embodiments, said mammal is a human. In some embodiments, said composition has a viscosity of from about 15 millipascal second (mPa·s) to 35 mPa·s at 37°C. In some embodiments, said composition has a viscosity of from about 20 millipascal second (mPa·s) to 30 mPa·s at 37°C. In some embodiments, said composition is injectable into a blood vessel of a mammal. In some embodiments, said mammal is a human. In some embodiments, said composition reduces blood flow within a blood vessel within a mammal when said composition is administered directly into the lumen of said blood vessel. In some embodiments, said composition blocks blood flow within a blood vessel within a mammal when said composition is administered directly into the lumen of said blood vessel. In some embodiments, tissue located outside of a blood vessel within a mammal and located within 20 cm of a blood occlusion site within said blood vessel is ablated when said composition is administered directly into the lumen of said blood vessel to form said blood occlusion site. In some embodiments, said tissue comprises cancer cells. In some embodiments, said tissue is selected from the group consisting of kidney tissue, liver tissue, brain tissue, prostate tissue, pancreas tissue, breast tissue, lung tissue, colon tissue, and bladder tissue. In some embodiments, said sugar is a sugar alcohol, a natural sugar, or a synthetic sugar; optionally, wherein said sugar is glycerol. In some embodiments, said sugar is selected from the group consisting of dextrose, fructose, galactose, glucose, lactose, maltose, polysorbates, sucrose, xylose, erythritol, glycerol, hydrogenated starch hydrolysates, isomalt, lactitol, maltitol, mannitol, sorbitol, xylitol, agave, honey, and molasses. In some embodiments, said composition is an embolic composition. In some embodiments, said composition is a catheter-directed embolic composition. Another aspect of the present disclosure related to a method for treating a solid tumor within a mammal, wherein said method comprises administering a composition into a lumen of a blood vessel within said mammal; wherein said composition forms at least one blood occlusion within said blood vessel at a location within 20 cm of said solid tumor; wherein said composition comprises an ionic liquid comprising geranic acid anions and choline cations; and a sugar; wherein the ratio of said geranate to said choline of said composition is from 6:1 to 1:6, optionally from 4:1 to 1:4, and further optionally from 3:1 to 1:3; and optionally wherein at least some cancer cells of said solid tumor are ablated. In some embodiments, said mammal is a human. In some embodiments, said solid tumor is a solid tumor located within a kidney, the liver, the brain, the prostate, the pancreas, a breast, a lung, the colon, or the bladder of said mammal. In some embodiments, said solid tumor comprises kidney cancer cells, liver cancer cells, brain cancer cells, prostate cancer cells, pancreatic cancer cells, breast cancer cells, lung cancer cells, colon cancer cells, or bladder cancer cells. In some embodiments, said composition comprises at least 0.1 percent of said ionic liquid, optionally at least 10 percent of said ionic liquid. In some embodiments, said composition comprises at least 25 percent of said ionic liquid. In some embodiments, said composition comprises at least 20 percent of said sugar, optionally at least 25 percent of said sugar. In some embodiments, said composition comprises wherein about 28 percent of said sugar. In some embodiments, the molar ratio of said geranic acid anions to said choline cations is from 1:3.5 to 1:6. In some embodiments, the molar ratio of said geranic acid anions to said choline cations is from 1:3.5 to 1:5.5. In some embodiments, the molar ratio of said geranic acid anions to said choline cations is from 1:3.5 to 1:5. In some embodiments, the molar ratio of said geranic acid anions to said choline cations is from 1:3.5 to 1:4.5. In some embodiments, the molar ratio of said geranic acid anions to said choline cations is from 1:3.5 to 1:4. In some embodiments, the molar ratio of said geranic acid anions to said choline cations is 1:4. In some embodiments, said composition comprises an imaging agent. In some embodiments, said composition comprises at least 10 percent of said imaging agent. In some embodiments, said imaging agent is selected from the group consisting of iohexol and tantelum. In some embodiments, said composition comprises a therapeutic agent. In some embodiments, said composition comprises at least 0.01 percent of said therapeutic agent. In some embodiments, the molecular weight of said therapeutic agent is greater than 7,500 Daltons. In some embodiments, the molecular weight of said therapeutic agent is greater than 10,000 Daltons. In some embodiments, the molecular weight of said therapeutic agent is greater than 15,000 Daltons. In some embodiments, the molecular weight of said therapeutic agent is greater than 25,000 Daltons. In some embodiments, the molecular weight of said therapeutic agent is greater than 50,000 Daltons. In some embodiments, the molecular weight of said therapeutic agent is greater than 100,000 Daltons. In some embodiments, the molecular weight of said therapeutic agent is greater than 125,000 Daltons. In some embodiments, the molecular weight of said therapeutic agent is less than 750,000 Daltons. In some embodiments, the molecular weight of said therapeutic agent is less than 500,000 Daltons. In some embodiments, the molecular weight of said therapeutic agent is less than 250,000 Daltons. In some embodiments, the molecular weight of said therapeutic agent is less than 200,000 Daltons. In some embodiments, said therapeutic agent comprises a biologically active polypeptide. In some embodiments, said biologically active polypeptide is selected from the group consisting of nivolumab, ipilimumab, pembrolizumab, atezolizumab, durvalumab, cemiplimab, and avelumab. In some embodiments, said therapeutic agent comprises an immunogenic polypeptide that is immunogenic within a mammal. In some embodiments, said mammal is a human. In some embodiments, said therapeutic agent comprises an antigen-binding polypeptide. In some embodiments, antigen-binding polypeptide is an antibody. In some embodiments, said antigen-binding polypeptide is selected from the group consisting of nivolumab, ipilimumab, pembrolizumab, atezolizumab, durvalumab, cemiplimab, and avelumab. In some embodiments, said therapeutic agent has anti-cancer activity within said mammal. In some embodiments, said therapeutic agent is selected from the group consisting of doxorubicin, cisplatin, paclitaxel, olaparib, everolimus, mitomycin, radioactive isotopes (e.g., yttrium Y-90, lutetium-177, actinium, fluorine-18, gallium-67, krypton-81m, rubidium-82, nitrogen-13, technetium-99m, indium-111, iodine-123, xenon-133, and thallium-201), atezolizumab, bevacizumab, cabozantinib-s-malate, ramucirumab, pembrolizumab, lenvatinib mesylate, sorafenib tosylate, nivolumab, pemigatinib, pembrolizumab, ramucirumab, regorafenib, and abemaciclib. In some embodiments, said composition comprises a nucleic acid. In some embodiments, said nucleic acid encodes an imaging agent. In some embodiments, said imaging agent is selected from the group consisting of blue fluorescent protein (BFP), green fluorescent protein (GFP), and red fluorescent protein (RFP). In some embodiments, said nucleic acid encodes a therapeutic agent. In some embodiments, said therapeutic agent encoded by said nucleic acid is selected from the group consisting of a biologically active polypeptide, an immunogenic polypeptide, and an antigen-binding polypeptide. In some embodiments, said therapeutic agent encoded by said nucleic acid comprises anti-cancer activity within said mammal. In some embodiments, said composition has a viscosity of from about 0.1 centipoise (cP) to about 1000 cP. In some embodiments, said composition has a viscosity of from about 15 centipoise (cP) to about 90 cP. In some embodiments, said composition reduces blood flow within said blood vessel. In some embodiments, said composition blocks blood flow within said blood vessel. In some embodiments, tissue located outside of said blood vessel and located within 20 cm of said blood occlusion site is ablated. In some embodiments, the composition is administered through catheterization, optionally using a microcatheter. In some embodiments, said sugar is a sugar alcohol, a natural sugar, or a synthetic sugar. In some embodiments, said sugar is selected from the group consisting of dextrose, fructose, galactose, glucose, lactose, maltose, polysorbates, sucrose, xylose, erythritol, glycerol, hydrogenated starch hydrolysates, isomalt, lactitol, maltitol, mannitol, sorbitol, xylitol, agave, honey, and molasses. In some embodiments, said sugar is glycerol. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although methods and materials similar or equivalent to those described herein can be used to practice the invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims. DESCRIPTION OF THE DRAWINGS Figures 1A – 1H. Characterization of ionic liquid embolic (ILE) formulations. Figure 1A) Various formulations of ILE indicating the ratio of geranic acid (GA) to choline (C). Figure 1B) Flow curves of ILEs at 37°C showing various viscosity levels with 4:1 being the most and 1:4 being the least viscous. Figure 1C) Injection force testing of ILEs. Figure 1D and 1E) Representative indocyanine green (ICG) and doxorubicin (DOX) fluorescence images in tissue-mimicking agar diffusion assay at 0, 4, and 24 hours. The diffusion coefficient was calculated based on diffusion area at 24 hour time-point. Figure 1F) Schematic of the TRANSWELL®diffusion assay conducted using ICG-incorporated ILEs (1:2, 1:3 and 1:4) and its effect on HepG2 cells. Figure 1G) Measurements of ICG fluorescence intensity in the lower chamber of TRANSWELL®inserts suggesting diffusion across the insert pores (400 µm) over 24 hours. Figure 1H) The viability assessment of HepG2 cells in the lower compartment of the TRANSWELL®inserts after 24 hours incubation. Figure 1I) Fractional viability of HepG2 cells treated with the ILEs and their corresponding IC50values are shown. Data are presented as mean ± s.e.m. (n = 4 for Figure 1C and Figure 1I; n = 3 for Figure 1D – 1F). Statistical significance was determined using one- way ANOVA (Figure 1C and Figure 1G), two-way ANOVA (Figure 1D and Figure 1E) followed by Tukey’s multiple comparisons test and nonparametric multiple t-tests (Figure 1H). ns, not significant; 0.0001. Figures 2A – 2H. Optimization of ILEs. Figure 2A) Viscosity measurements of ILEs mixed with various amounts of glycerol (0%, 10%, 20%, 30%, and 40%). Figure 2B and 2C) Fluoroscopic images (Figure 2B) and micro-CT images (Figure 2C) of ILE mixed with iohexol (IOH) and their corresponding intensity in X-ray fluoroscopy and micro-CT images of various concentrations of IOH (0%, 5%, 10%, 20%, 30%) and ILEs (1:2, 1:3, or 1:4)+glycerol (40% v / v)+IOH (20% v / v). Figure 2D) SDS-PAGE demonstrated the stability of nivolumab after mixing with various ILEs with or without glycerol for 24 hours at 37°C. Figure 2E) Zeta potential measurements of ILE, ILE+glycerol (40% v / v), and final ILE formulations containing ILE+glycerol+IOH+nivolumab+ICG. Figure 2F) Viscosity measurements of 3 candidates of ILE (ILE12, 13, and 14). Figure 2G) Break-loose and injection forces of ILEs injected through 2.8 French (F) 110 cm microcatheter at a constant flow rate of 1 mL / minute. Figure 2H) Fractional viability of HepG2 cells following incubation with neat ILE only, and final formulation of ILEs (ILE12, 13, and 14). Data are presented as mean ± s.e.m (n = 3 for Figure 2A – 2E; n = 4 for Figure 2F – 2H). Statistical significance was determined using one-way ANOVA (Figure 2A – 2F) and two-way ANOVA (Figure 2G) followed by Tukey’s multiple comparison test. ns, not significant; *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001. Figures 3A – 3L. In vivo characterization of ILEs in a nonsurvival porcine embolization model. Figure 3A) Baseline digitally subtracted angiography (DSA) showed normal arterial anatomy of the porcine kidney following contrast injection from a 5 French catheter in the main renal artery. Figure 3B) DSA following embolization of the lower pole arterial branches of a porcine kidney with ILE12, 13, or 14. Figure 3C) Gross images of resected kidneys at necropsy showed a discoloration of the lower half of the kidney demarcating the treated kidney tissue. Figure 3D) Kidneys in (Figure 3C) were transected along the sagittal plane and imaged using IVIS®imaging to detect the ICG; images localized the ICG in the ILE formulation to the treated lower lobe of the kidney. Figure 3E) Representative H&E images of embolized renal arteries. Figure 3F) Representative images of renal parenchyma stained with cleaved caspase 3. Figure 3G) Representative IHC stained for nivolumab showed detection of diffused nivolumab in the renal parenchyma, outside the arterial vasculature suggesting trans-arterial delivery. Figure 3H) Quantification of ICG intensities in the whole kidney indicated no significant difference between the three ILE formulations. Figure 3I) ICG radiance ratio of embolized (lower) and control (upper) kidney lobes. Figure 3J) Nuclei count of the vessel wall from the kidney tissue sections indicated uniform ablation of the vessel wall. Figures 3K and 3L) Quantitative analysis of cleaved caspase 3 (Figure 3K) and nivolumab (Figure 3L) by IHC showed significantly greater levels in ILE14 tissue as compared to ILE12 and ILE13. Data are presented as mean ± s.e.m (n = 8 for Figure 3H – 3J; n = 4 for Figure 3K and Figure 3L). Scale bars: Figure 3E, 50 µm; Figure 3F, 200 µm; and Figure 3G, 6 mm. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparison test. ns, not significant; **p < 0.01, ***p < 0.001 and ****p < 0.0001. Figures 4A – 4G. ILE14 in blood. Figure 4A) Schematic illustration of ILE14 endovascular injection from a microcatheter inside an artery demonstrating deep penetration into distal blood vessels. Figure 4B) Measurement of blood ACT with different concentrations of ILE (1:4) and ILE14 showing that >1% was able to delay thrombosis. Figure 4C) Representative hemorheology amplitude sweeps flow curves at 37°C after mixing with ILE (1:4) and ILE14. Figure 4D) Graph demonstrating storage modulus in blood samples at 37°C 30 minutes after mixing with ILE (1:4) and ILE14. Figure 4E) Hemolysis rate after incubation with various concentrations of ILE (1:4) and ILE14. Figure 4F) Lymphocyte count in porcine whole blood after incubation with different concentrations of ILE (1:4) and ILE14 indicating resistance to ablation. Figure 4G) Images of blood smear with Wright’s stain showed intact lymphocytes at 10% ILE (1:4) and ILE14. Data are presented as mean ± s.e.m (n = 3 for Figure 4B and Figure 4D; n = 5 for Figure 4E; n = 4 for Figure 4F). Figure 4C and Figure 4G: Representative result from n = 3. Scale bars: Figure 4G, 150µm and 25µm (inset). Statistical significance was determined using one-way ANOVA (Figure 4B and Figure 4D) and two-way ANOVA (Figure 4E and Figure 4F) followed by Tukey’s multiple comparison test. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001. Figures 5A – 5K. ILE14 in a survival porcine renal embolization model compared to clinically used bead-embolization. Figure 5A) Baseline and post-embolization (D+0) DSA of the porcine kidney. The upper lobe arteries of the kidney received the clinically used microbeads and the lower lobe arteries received ILE14. Both images indicate technically successful embolization. Figure 5B) DSA of control and embolized kidney at 7-day post-embolization. The upper lobe arteries that received microbeads demonstrated recanalization. The lower lobe arteries that received ILE14 demonstrated the persistent embolization of the arteries indicating that the effect was durable. Figure 5C) At necropsy, sagittal sections of the kidneys indicated greater atrophy in the lower lobe of the kidney that received ILE14 treatment (black arrow). Only the renal arteries embolized with ILE14 showed occlusion (red arrows). Figure 5D) IVIS®imaging of the kidneys in (Figure 5C) for ICG showed a persistent signal in the lower lobe of the kidneys that received ILE14 indicating delivery and persistence of the ICG suggesting that the ILE14 did not wash away. The upper pole of the kidney that received microbeads did not show any ICG signal suggesting that the ILE14 washed away. Figure 5E) Representative histology images of H&E and immunostaining for CD31, HIF-2α, cleaved caspase 3, and nivolumab. Images of slides from kidney tissue that received microbeads demonstrated a viable vessel wall, normal CD31 signal in the intima layer, and significant HIF-2α signal indicating a state of hypoxia. Cleaved Cas-3 showed mild, scattered evidence for apoptosis with beads detected in vessel walls (arrow) and the absence of any nivolumab detection. In contrast to the untreated control and the microbead-treated kidney sections, the ILE14- treated samples demonstrated complete embolization of the arteries and the absence of any CD31 signal, consistent with the absence of HIF-2α indicating that there is no viable vasculature. Corresponding cleaved Cas-3 signal showed diffuse apoptosis and uniform delivery of the nivolumab throughout the treated kidney tissue. Figure 5F) Quantitative analysis of ICG signal in the kidney 7 days post- embolization. Figure 5G) Nuclei count of the vessel wall from kidney sections. Figures 5H – 5K) Quantitation of CD31 (Figure 5I) HIF-2α (Figure 5J), cleaved caspase 3 (Figure 5K), nivolumab (Figure 5K) stained area. Data are presented as mean ± s.e.m (n = 8 for Figure 5F; n = 10 for Figure 5G; n = 5 for Figure 5H – 5K). Scale bars: Figure 5E, 200 µm (control), 75 µm (beads) and 150 µm (ILE14) for H&E, CD31 and HIF-2α staining.6 mm (control), 4 mm (beads) and 6 mm (ILE14) for cleaved caspase 3 and nivolumab staining. Statistical significance was determined using paired t-test (Figure 5F) and one-way ANOVA followed by Tukey’s multiple comparison test (Figure 5G). ns, not significant; **p < 0.01, ***p < 0.001 and ****p < 0.0001. Figures 6A – 6L. Evaluation of ILE14 in a rabbit VX2 liver tumor model. Figure 6A) Representative color ultrasound image of a rabbit VX2 liver tumor (dotted outline) showing blood flow. Figure 6B) Angiograms at baseline and post-ILE14 embolization to the segmental artery that feeds the rabbit tumor. The blue arrow indicates the tip of the microcatheter where ILE14 was slowly infused until stasis. The red arrow indicates the absence of blood flow in the embolized segment of the rabbit liver. Figure 6C) The gross image of the VX2 liver tumor post-embolization. Figure 6D) IVIS®imaging of the excised liver 1 hour post-embolization demonstrated specific ICG signal accumulation in a tumor region. Figure 6E) Sagittal transection of the tumor in (Figure 6D) showed the ICG signal predominantly localized to the tumor, in comparison to a control tumor that did not receive ILE14. Figure 6F) The quantitative analysis of total ICG radiance within the tumor tissue. Figure 6G) Representative H&E images of VX2 tumor showed tissue ablation and vessel occlusion in the ILE14 embolized tumor compared to control (arrows indicate tumor microvasculature). Figure 6H) Nuclei count within the tumor tissue sections. These images demonstrated trans-arterial diffusion of ILE14 and marked ablation of the adjacent cancer tissue. Figure 6I) Representative proliferating cell nuclear antigen (PCNA) immunostaining showed marked nivolumab delivery in the tumor that received ILE14. Figure 6J) Figure 6K) Quantification of PCNA positively stained area. Figure 6L) Quantification of nivolumab detection within the tumor tissues. Data are presented as mean ± s.e.m (n = 4 for Figure 6F; n = 3 (control) and n = 2 (embolized) for Figure 6H, 6J, and 6L)). Scale bars: Figure 6A, 1 cm. Figure 6G, 6I, 6K 75 µm. Statistical significance was determined using an unpaired t-test (Figure 6F, 6H, 6J, and 6L). *p < 0.05 and ***p < 0.001. Figure 7. Schematic illustration of ILE synthesis. Geranic acid was purified via a recrystallization process followed by the addition of choline bicarbonate. The mixture of geranic acid and choline bicarbonate metathesis reaction was prepared at room temperature (RT) until it no longer produced a CO2byproduct. Residual H2O was dehydrated using a rotary evaporator to create a neat ILE. Figures 8A – 8C. Characterization of ILE chemical structure. Figure 8A) Scheme of ILE synthesis. Figure 8B) The Fourier transform infrared (FTIR) spectra of geranic acid and choline bicarbonate with peak assignments. Figure 8C) The FTIR spectra of ILE synthesized using 7 different molar ratios of geranic acid and choline bicarbonate. Figures 9A and 9B. Characterization of ILE+glycerol. Figure 9A) The FTIR spectrum of glycerol. Figure 9B) The FTIR spectrum of ILE with or without glycerol (40% v / v). Adding glycerol to ILE yielded an increase in C-O peaks at 1000-1150 cm-1. Figure 10. Evaluation of small molecule diffusivity loaded in glycerol-incorporated ILEs in a tissue-mimicking agarose matrix. Fluorescence imaging of ICG and DOX diffusion in 2% agarose assay constructs and quantitative analyses of diffusivity after mixing with three different molar ratios of ILE formulations with or without glycerol. The results showed enhancement of radial diffusion compared to free ICG and a stepwise increase in diffusion with a higher choline ratio in ILE at 24 hours (top) whereas DOX diffusion was similar for all formulations (bottom). Quantitative analysis demonstrated that the addition of glycerol did not hinder the diffusivity of ICG or DOX. Data are presented as mean ± SEM (n= 3) Statistical significance was determined using two-way ANOVA followed by Tukey’s multiple comparisons test. ns, not significant; **p < 0.01 and ***p < 0.001. Figures 11A and 11B). Long-term stability of nivolumab in ILEs. Figure 11A) Representative SDS-PAGE performed under non-denaturing conditions showed a stable protein band at 146 kDa of nivolumab incubated in saline (control) or with different molar ratios of ILE or ILEs containing glycerol at 37°C up to 28 days. Figure 11B) Quantitative analysis demonstrated similar nivolumab band intensity to control in all formulations up to 28 days. Data are presented as mean ± SEM (n= 3). Statistical significance was determined using one-way ANOVA and Tukey’s post-hoc multiple comparison tests. ns, not significant. Figure 12. The stability of ICG in different ILE formulations. Fluorescence images of opaque wells loaded with aliquots of ICG dissolved in saline (control), or mixed with different ILE formulations showed a rapid loss of ICG fluorescence within 24 hours in the control wells, compared to sustained high signal in the presence of glycerol in ILE (1:2)+gly+ICG, ILE (1:3)+gly+ICG, or ILE (1:4)+gly+ICG up to 28 days. The addition of an iohexol contrast agent, and the checkpoint inhibitor, nivolumab (ILE12, 13, and 14), did not impact the ICG signal. Data are presented as mean ± SEM (n= 3). Statistical significance was determined using one-way ANOVA and post-hoc Tukey’s multiple comparisons tests. ns, not significant; *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001. Figure 13. In vitro protein release profile of ILE. Graphs of cumulative protein release profile of ILE14 with nivolumab (146 kDa) or BSA (67 kDa) loaded into dialysis tubes with a pore size of 300 kDa and incubated at 37°C. Initial burst release was seen within the first 24 hours for both proteins while BSA showed subsequent sustained release behavior up to day 7. Figure 14. Renal artery embolization with ILE in swine. Fluoroscopic imaging showed an angiographic microcatheter positioned within the renal artery during ILE embolization. ILE formulations contained an iohexol contrast agent to enable real-time visualization of the embolic material during the procedure. Visualization of radiodense ILE exiting the tip of the microcatheter and distributing throughout distal branches was observed (arrows). Figures 15A – 15D. Assessing the efficacy of renal artery embolization using ILE12 in non- survival pigs. Figure 15A) Baseline DSA showed patency of the porcine renal arteries at baseline prior to embolization. Figure 15B) DSA immediately following transcatheter injection of ILE12 through the lower pole renal artery branches showed successful embolization in the lower pole. Figure 15C) Gross view of explanted kidneys at 1 hour after embolization. Figure 15D) Fluorescence imaging of transected kidneys showed high ICG fluorescence throughout the lower pole cortex in all kidneys that were embolized with ILE12. Figures 16A – 16D. Assessing the efficacy of renal artery embolization using ILE13 in non- survival pigs. Figure 16A) Baseline DSA showed patency of the porcine renal arteries at baseline prior to embolization. Figure 16B) DSA immediately following transcatheter injection of ILE13 through the lower pole renal artery branches showed successful embolization in the lower pole. Figure 16C) Gross view of explanted kidneys at 1 hour after embolization. Figure 16D) Fluorescence imaging of transected kidneys showed high ICG fluorescence throughout the lower pole cortex in all kidneys that were embolized with ILE13. Subject 2, left kidney demonstrated ICG signal in the upper pole due to aberrant blood vessels arising from the lower pole arteries. Figures 17A – 17D. Assessing the efficacy of renal artery embolization using ILE14 in non- survival pigs. Figure 17) Baseline DSA showed patency of the porcine renal arteries at baseline prior to embolization. Figure 17B) DSA immediately following transcatheter injection of ILE14 through the lower pole renal artery branches showed successful embolization in the lower pole. Figure 17C) Gross view of explanted kidneys at 1 hour after embolization. Figure 17D) Fluorescence imaging of transected kidneys showed high ICG fluorescence throughout the lower pole cortex in all kidneys that were embolized with ILE14. Additional samples demonstrated ICG signal in the upper pole due to aberrant blood vessels arising from the lower pole arteries (arrows). Figures 18 A – 18L. Renal artery embolization using ILE (1:4) or glycerol only in non-survival porcine renal embolization model. Figure 18A) Baseline DSA showed normal renal arteries before embolization. Figure 18B) DSA following embolization of the lower lobe renal arteries with ILE (1:4; without glycerol) demonstrated successful occlusion of the treated arteries. Injection of glycerol only into the lower pole renal arteries demonstrated rapid washout from the renal arteries indicating that glycerol alone did not achieve embolization. Figure 18C) Gross view of the explanted kidneys during necropsy demonstrated a normal appearance of the kidney that received glycerol and mild discoloration of the lower half of the kidney that received ILE. Figure 18D) Fluorescence imaging of transected kidneys showed ICG fluorescence distribution throughout the lower pole cortex in the kidney that was embolized with ILE (1:4), whereas embolization with glycerol resulted in a weak ICG fluorescence. Figure 18E) Representative H&E cross-sectional images of renal arteries that received ILE (1:4) or glycerol showed an embolus incompletely casting the artery lumen with viable endothelial cells (arrows) and a widely patent lumen of the renal artery that was injected with glycerol suggesting recanalization. Figure 18F) Representative IHC images stained for cleaved caspase-3 showed minimal signs of apoptosis in the renal cortex after embolization with ILE (1:4) or glycerol. Figure 18G) Representative IHC images showed minimal nivolumab detection in the renal parenchyma. Figure 18H) Quantitative analysis of ICG fluorescence ratio (inferior pole: superior pole) showed significantly lower fluorescence in the kidneys that received embolization with glycerol (suggesting washout) compared to higher fluorescence in the ILE (1:4)-embolized kidney. Figure 18I) Quantitative analysis of ICG fluorescence in the whole kidney. Figure 18J) Morphometric analysis of nuclei count in the arterial wall of the renal artery branches showed the effect of embolization using ILE (1:4) and glycerol compared to non-embolized control vessels. Figure 18K) Quantitative analysis of cleaved caspase-3 in the renal tissue sections at one hour after embolization with ILE (1:4) or glycerol. Figure 18L) Quantitative analysis of nivolumab immunostaining in the kidney sections at one hour after embolization with ILE (1:4) or glycerol. Data are presented as mean ± SEM (n = 8 for ILE 1:4 and n = 6 for glycerol for Figure 18H and Figure 18I; n = 8 for Figure 18J; n = 4 for ILE 1:4 and n = 3 for glycerol for Figure 18K and 18L). Scale bars: Figure 18E, 50 µm; Figure 18F, 200 µm; and Figure 18G, 4 mm. Statistical significance was determined using an unpaired t-test (Figure 18H, 18I, 18 K, and 18L) and one-way ANOVA followed by Tukey’s multiple comparison test (Figure 18J). ns, not significant; *p < 0.05, ***p < 0.001 and ****p < 0.0001. Figure 19. Recanalization of vessels after embolization with glycerol. Figure 20. Transcatheter injection of ILE (1:4) for renal embolization in a nonsurvival porcine model. Upon confirming the patency of renal arteries using DSA, ILE (1:4) was delivered into the lower lobe for embolization. Gross images of explanted kidneys 1 hour after embolization showed discoloration of the lower half of the kidneys that received ILE (1:4) injection. IVIS®imaging of transected kidneys showed ICG distribution only in the lower lobe indicating delivery of ILE (1:4). Figure 21. Transcatheter injection of glycerol for renal embolization in a nonsurvival porcine model. Upon confirming the patency of renal arteries using DSA, glycerol was delivered into the lower lobe for embolization. Post-embolization DSA showed immediate recanalization and normal appearance of the kidneys during necropsy. IVIS®imaging of transected kidneys showed minimal ICG signal consistent with recanalization observed during the procedure. Figures 22A and 22B. Assessment of blood oxidation in various concentrations of ILE (1:4) and ILE14. Figure 22A) The absorbance spectrum of blood incubated with 10% ILE (1:4) displayed the methemoglobinemia (MetHb) spectrum. Figure 22B) The absorbance spectrum of blood incubated with 10% ILE14 displayed oxyhemoglobin (OxyHb) spectrum. Figure 23. Hemocompatibility of ILE (1:4), glycerol, and ILE14. Graphs demonstrating the effect of different dilutions of ILE (1:4), glycerol, and ILE14 on white blood cells, monocytes, granulocytes, and platelets count. Data are presented as mean ± SEM (n= 4). Statistical significance was determined using two-way ANOVA followed by Tukey’s multiple comparison test. ns, not significant, *p < 0.05 and ****p < 0.0001. Figure 24. A microscopic image of clinically used microbeads (BeadBlock™) measuring 100- 300 µm in diameter that are typically used during embolization procedures in humans and were tested in the survival porcine renal artery embolization model. Figures 25A – 25E.7-day survival studies following embolization of renal arteries in swine with microbeads and ILE14. Figure 25A) DSA images showed normal renal arteries before embolization. Figure 25B) DSA images post-embolization; kidney 1 only received ILE14 and in kidneys 2-5 the lower pole arteries received ILE14 and the upper pole received microbeads at Day=0. All renal arteries showed technically successful embolization. Figure 25C) DSA images at Day=7 demonstrated absence of any perfusion in kidney 1 that received ILE14 only. In kidneys 2-5, all upper pole arteries that received microbeads demonstrated recanalization; all lower pole arteries that received ILE14 were persistently occluded. Figure 25D) Gross images of kidneys during necropsy showed evidence of ischemia in the poles that received ILE14. Figure 25E) Fluorescence imaging of ICG distribution in the bisected kidneys at 7 days after embolization showed significant fluorescence in all of kidney 1 and only in the lower half of the kidneys 2-5 that received ILE14. The absence of significant ICG detection in the upper pole of the kidneys was consistent with recanalization seen in Figure 25C. Figure 26. Renal embolization with clinically used microbeads. Representative H&E histology section showed a microbead partially occluding a renal artery branch (arrow) with approximately 400 µm size whereas smaller vessels appeared patent suggesting the difficulty of achieving complete occlusion due to variations in the shape and dimensions of the vasculature (asterisk). Scale bar: 250 µm. Figure 27. Complete blood count and biochemical analysis of pig serum after renal artery embolization with ILE14. Complete blood count and analyses for serum markers of systemic injury demonstrated no significant differences at 1 hour and 7 days after embolization compared to baseline. Data are presented as mean ± SEM (n= 5). Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparison test. ns, not significant, *p < 0.05 and ***p < 0.001. Figure 28. Histological evaluation of various organs following ILE14 renal artery embolization in swine at Day=7. Stained histology sections showed no noticeable changes in morphology. Scale bars: 250 µm. Figure 29. Theoretical physiologic concentration of ILE14 after performing embolization. Figure 30. Migration assay using sub-lethal doses of ILE14. Sub-lethal doses of ILE14 were used to assess whether it can affect the migration capability of HepG2 cells. The results showed that even at concentrations that did not affect viability, ILE14 was able to impede the proliferation of HepG2 cancer cells. Data are presented as mean ± SEM (n= 3). Scale bars: 500 µm. Statistical significance was determined using a two-way ANOVA test. *p < 0.05 and ****p < 0.0001. Figures 31A – 31D. Image-guided embolization of the rabbit liver bearing a VX2 tumor using ILE14. Figure 31A and 31B) DSA images from two rabbits showed a 2.4 F microcatheter positioned inside the hepatic artery pre- and post-embolization with ILE14. Figure 31C) Gross images of exposed livers showed ICG in the VX2 tumors at one-hour post-embolization with ILE14. Figure 31D) IVIS®images of embolized VX2 liver tumor. Figures 32A and 32B. Histological evaluation of VX2 tumor tissues at 1 hour after embolization with ILE14. Figure 32A) Representative H&E-stained histology sections of VX2 tumors showing embolized vessels (arrows) surrounded by ablated tissues throughout the treated zone. Figure 32B) Immunostaining for PCNA showing a marked decrease in PCNA detection in the perivascular ablated areas. Scale bars: 150 µm. Figure 33. Localization of nivolumab in the VX2-bearing rabbit liver after embolization with ILE14. Immunostained VX2 bearing liver section showed extensive detection of nivolumab after embolization with ILE14 demonstrating the ability of ILE to induce efficient transvascular diffusion of immunotherapy throughout the tumor zone and the adjacent area. Figure 34. Immunostaining of nivolumab in VX2 tumor after embolizing adjacent hepatic segment. Immunostained tissue section harvested from a rabbit liver bearing a VX2 tumor after embolization of an adjacent hepatic segment with ILE14 showed the absence of nivolumab within the tumor zone suggesting that only embolization of the targeted tumor arteries leads to ablation and drug delivery. Scale bars: Tiled image (top); 3 mm and inset (bottom); 400 µm. Figures 35A – 35E. Injection of ILE14 into human solid tumors. Figure 35A) Representative ex vivo tumor tissue specimen harvested from a patient with renal cancer that was injected with ILE14 demonstrated time-dependent enhancement of ICG fluorescence and diffusion area up to 24 hours. Figure 35B and 35C) Quantitative analysis of radiant efficiency and diffusion area respectively at 1, 12, and 24 hours after the injection. Figure 35D) Images of H&E-stained histology sections and graphic summary of nuclei count demonstrated marked ablation and complete elimination of nuclei in the ILE14 injected renal cancer tissues compared to the untreated samples. Figure 35E) Immunostaining for nivolumab and morphometric quantification of positively stained tissue area of control and ILE14-injected human tumor tissues. Scale bars: Figures 35D and 35E, 200 µm. Statistical significance was determined using an unpaired t-test (Figures 35D and 35E) and one-way ANOVA followed by Tukey’s post hoc multiple comparison tests (Figures 35B and 35C). ns, not significant; *p < 0.05, **p < 0.01 and ****p < 0.0001. Figure 36. Antibacterial property of ILE14. Heat map illustrating antimicrobial susceptibility of a serially diluted ILE14 against common pathogens associated with nosocomial infections including Escherichia coli (E. coli), the Methicillin-Resistant Staphylococcus aureus (MRSA), Vancomycin- Resistant Enterococci (VRE), Carbapenem-Resistant Enterobacteriaceae (CRE), Clostridium difficile (C. difficile), and the multidrug-resistant Candida auris (C. auris). Figures 37A – 37B. Limited access of OBSIDIO™conformable gel embolic in the distal microvessels. Figure 37A) DSA in a 63-year-old male with acute and severe hemoptysis secondary to lung adenocarcinoma. Circle indicates the area of hemorrhage. The patient underwent pulmonary arterial embolization using OBSIDIO™gel embolic, resulting in instant hemostasis. The gel embolic successfully embolized the upstream larger bronchial artery and did not address the smaller bleeding blood vessels distally. Figure 37B) The computed tomography (CT) chest angiogram during the 6-day follow-up revealed a stable OBSIDIO™gel embolic but confirmed that the peripheral microvasculatures were not directly targeted. A few days after embolization, bleeding resumed from the distal microvessels requiring a surgical resection. Figures 38A – 38J. Characterization and mechanism of P-LE. Figure 38A) The formulation of P- LE consists of IL, PEG (100 kPa), and IOH. Figure 38B) Hemorheology curves of blood mixed with IL generated in different stoichiometric ratios. Figures 38C – 38E) Activated clotting time (ACT) of platelet poor plasma (Figure 38C), platelet rich plasma (Figure 38D), and red blood cells (Figure 38E), when mixed with IL (- / +) or calcium chloride (CaCl2) (- / +) (n=3). Figure 38F) Representative hemorheology curves of various concentrations of RBC (0, 12.5, 25, 50, and 100%) mixed with IL at 37℃. Figure 38G) Storage modulus (G′) of the mixture of IL and various amounts of RBC (0, 12.5, 25, 50, and 100% v / v) (n=3). Figures 38H – 38J) Blood clotting index (BCI) of citrated whole blood (Figure 38H), heparinized whole blood (Figure 38I), and defibrinated whole blood (Figure 38J) mixed with IL (- / +) or CaCl2(- / +) (n=6). Data are mean ± s.e.m.. Statistical analysis was performed using one-way ANOVA. ** p < 0.01 and **** p < 0.0001. Figures 39A – 39I. Optimization of P-LE. Figure 39A) The experimental setup for measuring the displacement pressures. The vessel occlusion was simulated inside a silicone tube and blood was infused at a flow rate of 30 mL / minute with a syringe pump. Figure 39B) The quantification of maximum pressure required to displace control (anticoagulated blood), a blood clot (anticoagulated blood+CaCl2), and anticoagulated blood mixed with IL (n=3). Figure 39C) Viscosity curves of IL, PEG, and IL+PEG characterized by shear rate sweeps. Figure 39D) A hemorheology curves of anticoagulated blood mixed with PBS (control), IL, or IL+PEG at 37℃. Figure 39E) Fluoroscopic images of IL+PEG mixed with various concentrations of IOH (0, 5, 10, 20, and 30% v / v) and their corresponding intensities (n=5). Figure 39F) A hemorheology curve of anticoagulated blood mixed with P-LE at 37℃. Figure 39G) An injection force of P-LE to inject through a 2.8F microcatheter (length: 100 cm). Figure 39H) Representative displacement pressure curves of various simulated vessel occlusions. Figure 39I) The quantification of maximum pressure required to displace control (anticoagulated blood), a blood clot (anticoagulated blood+CaCl2), anticoagulated blood mixed with IL, PEG, or P-LE (n=3). Data are mean ± s.e.m.. Statistical analysis was performed using one-way ANOVA. * p <0.05, and **** p < 0.0001. Figures 40A – 40O. PEG, IL, or P-LE injection in rat femoral artery embolization model. Figure 40A) Rat femoral artery embolization performed with PEG only. Severe breakthrough bleeding from the needle-puncture hole was observed post-injection, suggesting that the PEG does not obstruct blood flow when used alone. Figure 40B) Representative H&E images of untreated and PEG-injected FA. PEG- injected arteries appear to be normal with no sign of occlusion. Scale bars: 500 µm. Figure 40C) The quantification of vascular nuclei demonstrates no significance between control and PEG groups (n=3). Figure 40D) Rat FA embolization performed with IL alone. Minor breakthrough bleeding from the needle-puncture hole was observed post-injection of IL, suggesting that IL-alone injection resulted in an incomplete occlusion of FA (the asterisk shows the breakthrough bleeding). Laser speckle contrast imaging (LSCI) showed minimal perfusion change in FA pre- and post-embolization. Figure 40E) Representative hindlimb LSCI images showing minimal changes of perfusion after embolization with IL. Figure 40F) The quantification of hindlimb perfusion rate in control and embolized hindlimb before and after IL injection (n=3). Figure 40G) Micro-CT image showing poor embolization in FA and its downstream microvessels; the majority of IOH has been washed out. Figure 40H) Representative H&E and CD31 immunostaining images of control and IL-injected FA. Scale bars: 500 µm. Figure 40I) The quantitative analysis of vascular nuclei and CD31-positive cells of IL-injected FA (n=3). Figure 40J) Gross and LSCI analyses of rat FA embolization showing the persistent embolization using P-LE (yellow arrows indicate the created embolus). Figure 40K) LSCI images of control and embolized rat hindlimbs pre- and post-embolization with P-LE. Figure 40L) Hindlimb perfusion rate change of control and embolized limbs compared to respective baseline measurements (n=3). Figure 40M) Micro-CT image showing the embolization of rat FA and its downstream microvessels with P-LE. Figure 40N) Representative H&E and CD31 immunostaining images of untreated and embolized FA with P-LE. Scale bars: 250 µm. Figure 40O) Average count of vessel wall nuclei and CD31+cells per vessel for control and embolized FA (n=3). The images shown are representative of the 3 mice analyzed per group. Data are mean ± s.e.m.. Statistical analysis was performed using an unpaired t-test in Figures40C, 40F, and 40L, and multiple unpaired t-tests in Figures 40I and 40O. ns, not significant; ** p < 0.01 and *** p < 0.001. Figures 41A – 41F. P-LE injection in porcine renal embolization non-survival model. Figure 41A) Representative DSA images showing arterial anatomy of a porcine kidney before injecting P-LE (baseline), during the injection of P-LE to the lower lobe of the kidney, and post-embolization (arrows indicate the site of P-LE injection). Figure 41B) Gross images of whole and bisected kidneys showing discoloration in the lower lobe embolized with P-LE demarcating the area of injection. Scale bars: 3 cm. Figure 41C) IVIS image of a bisected kidney showing ICG signals localized in the lower renal cortex region, and the quantification of ICG radiance in control (non-treated lobes) and P-LE-embolized lobes (n=4). Figure 41D) H&E image of renal cortex embolized with P-LE and its representative magnified images showing embolized microvessels (top). Relative frequencies of embolized vessels in various diameters were quantified (n=300). Scale bars: 200 µm. Figure 41E) Representative H&E and CD31 immunostaining images of vessels in control and embolized kidneys. Scale bars: 100 µm. Figure 41F) The count of total cells in vessel walls and CD31+cells per vessel in control and embolized kidneys (n=10). The images shown are representative of the 4 pigs analyzed per group. Data are mean ± s.e.m.. Statistical analysis was performed using an unpaired t-test in Figure 41C, and multiple unpaired t-tests in Figure 41F. ** p < 0.01 and **** p < 0.0001. Figures 42A – 42H. P-LE injection in heparinized canine renal embolization non-survival model. Figure 42A) Representative DSA images showing arterial anatomy of a heparinized canine with no embolization (control) and pre- and post-embolization with P-LE. Figure 42B) IVIS image of a bisected kidney showing ICG fluorescence from P-LE formulation localized in the renal parenchyma after embolization. Figure 42C) Gross image of a bisected kidney and its representative magnified images showing the embolized vessels (indicated arrowheads). Scale bars: 2 cm. Figure 42D) H&E images of whole tissue sections and their magnified images of kidney parenchyma in control and embolized kidneys. Arrowheads indicate microvessels in control and embolized kidneys, respectively. Scale bars: 6 mm (top) or 1 mm (bottom). Figure 42E) Representative H&E and CD31 immunostaining images of vessels in control and P-LE-injected kidneys. Scale bars: 100 µm. Figures 42F – 42H) Average nuclei count in vessel walls (Figure 42F) and perivascular regions (Figure 42G), and CD31+ cells (Figure 42H) per vessel in non-embolized (control) and embolized kidneys (n=12). The images shown are representative of the 4 canines analyzed per group. Data are mean ± s.e.m.. Statistical analysis was performed using an unpaired t-test in Figures 42F – 42H. **** p < 0.0001. Figures 43A – 43G. P-LE injection in porcine renal embolization survival model. Figure 43A) Baseline and post-embolization (D0 and D14) DSA of a porcine kidney. The upper pole of the kidney was injected with clinically used 300 micron microbeads (EMBOSPHERE®) and the lower pole was injected with P-LE. Both showed successful initial embolization (D0), but the upper lobe injected with microbeads was recanalized at D14 while P-LE showed persistent embolization. Figure 43B) Gross and IVIS images of a bisected kidney showing discoloration and ICG localization in the lower pole of the kidney demonstrating permanent embolization by P-LE. Scale bars: 3 cm. Figure 43C) Representative H&E images of renal cortex parenchyma in beads and P-LE-injected kidneys. Arrowheads indicate microvessels in beads and P-LE-injected kidneys, respectively. Scale bars: 500 µm. Figure 43D) Relative frequencies of embolized microvessels. Only 14 vessels were shown to remain occluded with beads indicating that the majority was patent. Figure 43E) Representative H&E, CD31, and myeloperoxidase (MPO) immunostaining and trichrome images of microvessels suggesting persistent embolization of microvessels with P-LE. Scale bars: 50 µm. Figure 43F) Quantitative analysis of vascular nuclei count and CD31 immunostaining from microvessels show significant differences with P-LE embolization. Quantification of MPO immunostaining shows no significance among all tested groups. Quantitative analysis of collagen fraction in microvessels demonstrated significantly lower collagen content in P-LE- embolized vessels indicating persistent occlusion of vessels (n=12). Figure 43G) Fluoroscopic images of hemorrhaging microvessels and post-embolization with P-LE. Immediate hemostasis was achieved with P-LE embolization. The images shown are representative of the 6 pigs analyzed per group. Data are mean ± s.e.m.. Statistical analysis was performed using two-way ANOVA in Figure 43F (left) and one-way ANOVA in Figure 43F (right). ns, not significant; ** p < 0.01 and **** p < 0.0001. Figure 44. The tilt test was conducted 5 minute after mixing various IL+PEG with anticoagulated blood to grossly assess the blood gelation. The results indicate that at a PEG concentration of 100 mg / mL, the gelation process was faster, with no signs of flow. Figure 45. ACT of citrated whole blood when mixed with various permutations of IL (- / +), PEG (- / +), or CaCl2(- / +) (n=3). Data are mean ± s.e.m.. Statistical analysis was performed using two-way ANOVA. ** p < 0.01 and **** p < 0.0001. Figures 46A – 46B. Figure 46A) Images of blood thrombosis test using anticoagulated citrated blood mixed with PBS (control), P-LE, Coil, or Coil+P-LE at 37℃ taken at different time points (1-30 minute). Figure 46B) Images showing the rapid formation of thrombus inside the coil when P-LE was mixed with anticoagulated blood suggesting the rescue of coil embolization in clinical scenarios. Figure 47. Images showing the procedure of embolization in a rat femoral artery (FA). After ligation, materials were injected into the FA, filling the whole lumen. The proximal ligature was immediately removed to allow blood influx and mixing with P-LE. The distal ligature was subsequently removed to evaluate embolization and resulting hemostasis. Figure 48. The comparison between OBSIDIO™and P-LE in a rat femoral artery embolization model. While OBSIDIO™mostly remained upstream, P-LE readily reached and embolized sub-mm downstream vessels. Scale bars: 1 mm. Figures 49A – 49G. Figure 49A) ACT analysis of normal (pre-heparinization) blood and heparinized blood of rats (n=3). Figure 49B) LSCI analysis of FA embolization in heparinized rats showing durable embolization with P-LE (arrows indicate created embolus). Figure 49C) LSCI images of control and embolized rat hindlimbs in a heparinized rat pre- and post-embolization with P-LE. Figure 49D) Hindlimb perfusion rate change of control and embolized limbs compared to baseline in heparinized rats (n=3). Figure 49E) Micro-CT image showing the embolization of FA and its downstream microvessels in a heparinized rat. Figure 49F) Representative H&E and CD31 immunostaining images of non-embolized FA (control) and FA embolized with P-LE in a heparinized rat. Figure 49G) Average count of vessel wall nuclei and CD31+cells per vessel of control and embolized FA in heparinized rats (n=3). Data are mean ± s.e.m.. Statistical analysis was performed using an unpaired t-test in Figure 49A and Figure 49D, and multiple unpaired t-tests in g. * p <0.05 and ** p < 0.01. Figure 50. A gross image of the respective bisected porcine kidney without embolization (control) and post-embolization (embolized). Figure 51. HIF-2α immunostaining images of the upper lobe of a porcine kidney which was untreated (control) and the lower lobe which was embolized with P-LE (embolized) showing hypoxia regions surrounding the embolized vessels. Figure 52. Angiogram of spleen before and after embolization using P-LE. Post-embolization, the splenic sinusoids are no longer visible. Figure 53. ACT of normal (baseline) and heparinized canine whole blood showing successful anticoagulation with heparin (n=3). Data are mean ± s.e.m.. Statistical analysis was performed using an unpaired t-test in c. ** p < 0.01. Figure 54. DSA, gross images, cross-section images, and IVIS images of a non-embolized porcine kidney (control lobe) and an embolized porcine kidney (treated lobe). The upper pole was injected with microbeads and the lower pole was injected with P-LE. The only area injected with P-LE showed evident discoloration at D14 suggesting persistent embolization. IVIS imaging taken at D14 demonstrated intense ICG fluorescence while the pole injected with the bead barely retained the ICG signal suggesting a washout. Figure 55. MPO immunostaining of renal capsule of the tissues injected with beads and P-LE. Beads-injected tissue appears to be normal with cellularity and no sign of MPO. P-LE-injected tissue demonstrates no detectable nuclei in the parenchyma with infiltrated inflammatory cells only in the renal capsule region secondary to separate blood supply of the renal capsule from the capsular artery. Figures 56A – 56D. Oxygen saturation (SpO2) (Figure 56A), ACT (Figure 56B), complete blood cell (CBC) counts (Figure 56C), and serum biochemical analysis (Figure 56D) results from baseline, D0 post-embolization, and D14 post-embolization blood samples in a survival porcine renal embolization model (n=6). Data are mean ± s.e.m.. Statistical analysis was performed using one-way ANOVA in Figures 56A and 56B, and two-way ANOVA in Figures 56C and 56D. ns, not significant; * p < 0.05 and ** p < 0.01. Figures 57A – 57B. Figure 57A) Antimicrobial effect of P-LE against patient-derived antibiotic- resistant pathogens (MRSA, CRE, VRE, C. difficile, and C. auris). P-LE was serially diluted with each microbe, incubated overnight, and inoculated on blood agar plates to examine the minimum inhibitory (MIC) concentration of P-LE. The susceptibility for each tested microbe is summarized in the table. Figure 57B) Antimicrobial effect of 210 and 211-fold diluted P-LE against MRSA and C. auris. Figures 58A – 58K. Preparation and characterization of ionic liquids (ILs) and optimization of LEAD formulation. Figure 58A) Exemplary chemical reaction yielding ILs and image of the synthesized IL3:1, IL1:1, IL1:3 formulations in glass vials. Figure 58B) Exemplary schematic representation of the chemical reaction yielding ILs. Figure 58C) Representative fluorescence images and a graph demonstrating ICG diffusion from IL preparations, with IL3:1 exhibiting the largest diffusion area. Figure 58D) Fractional viability assessment of several human cancer cell lines (SNU478, HUCCT1, HEPG2) for IL3:1. Figure 58E) Live / dead cell viability assay of SNU478 cells incubated with different concentrations of IL3:1, indicating a dose-dependent cancer cell death (green) compared to control group. Figure 58F) Gel electrophoresis and graph showing the stability of Nivolumab band in IL3:1 solution up to day 28, compared to Nivolumab in saline (control). Figure 58G) LEAD formulation (70% IL3:1 + 20% IOH + 1 mg / mL Nivolumab + 0.25 mg / mL ICG) displaying adequate visibility under fluoroscopic imaging. Figure 58H) Diffusion assay comparing IL3:1 and LEAD, indicating no difference in ICG diffusion. Figure 58I) Fractional viability assessment in the SNU478 cancer cells for IL3:1 and LEAD, revealing comparable cytotoxicity. Figure 58J) Measurements of break loose and injection force for LEAD, demonstrating ease of injectability with a microcatheter. Figure 58K) Viscosity measurement of LEAD. Statistical differences were calculated using unpaired T-Test, or ANOVA and Tukey’s post- hoc tests as applies. Data in all graphs are the means ± SEM (n ≥ 3). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. Figures 59A – 59G. In vitro evaluation and anti-bacterial activity of LEAD. Figure 59A) An illustration detailing the transwell assay design, employing LEAD or saline + ICG overlayed on top of 2% agarose gel and porcine blood layers. The graphs depict the cytotoxic effect and enhanced drug diffusion through tissue-like barriers of LEAD. Figure 59B) Microscopy and quantitative analysis of SNU478 cell migration at 24 hours post-treatment with 0% (control), 0.31%, and 0.63% LEAD, revealing reduced cell migration. Figure 59C) Microscopy and quantitative analysis of SNU478 cell adhesion at 4 hours post-treatment with 0% (control), 0.31%, and 0.63% LEAD, indicating decreased cell adhesion. Figure 59D) Sterility assessment of LEAD following a 1-day incubation at 37°C. Figure 59E) Antibacterial evaluation against E. coli using serially diluted concentrations of LEAD, demonstrating a potent antibacterial effect. Figure 59F) Representative blood agar plates of antibiotic-resistant patient- derived bacterial strains after incubation in serially diluted LEAD solutions. Numbers (n) in the figure indicate the LEAD dilution factor (2n). Figure 59G) Table summarizing the susceptibility of antibiotic- resistant patient-derived bacterial strains to LEAD. Statistical differences were calculated using unpaired T-Test, or ANOVA and Tukey’s post-hoc tests as applies. Data in all graphs represent means ± SEM (n ≥ 3). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. Figures 60A – 60J. Tissue ablation and drug delivery using LEAD in ex vivo human liver tissues. Figure 60A) Representative gross image of a non-fibrotic human liver tissue before LEAD injection. Figure 60B) IVIS images of the representative human liver tissue at baseline, 1-hour, and 24-hour post- injection, with a graph depicting changes in ICG fluorescence intensity over time. Figure 60C) High- power microscopic images of H&E and Nivolumab-immunostained of non-fibrotic human liver tissue sections after LEAD injection or control. Figures 46D – 46E) Quantitative analysis of cell density (Figure 60D) and nivolumab detection area (Figure 60E) in non-fibrotic human liver tissues. Figure 60F) Representative gross image of a fibrotic human liver tissue before LEAD injection. Figure 60G) IVIS images of the representative human fibrotic liver tissue at baseline, 10 minutes, 1-hour, and 24-hour post- injection, with a graph depicting changes in ICG fluorescence intensity over time. Figure 60H) High- power microscopic images of H&E and nivolumab-immunostained of fibrotic human liver tissue sections after LEAD injection or control. Figures 60I – 60J) Quantitative analysis of cell density (Figure 60I) and nivolumab detection area (Figure 60J) in fibrotic human liver tissues. Statistical differences were calculated using unpaired t-tests. Data in all graphs represent means ± SEM (n ≥ 3). ***P < 0.001, and ****P < 0.0001. Figures 61A – 61G. In vivo portal vein embolization, ablation, and drug delivery in a rat liver model. Figures 61A – 61B) Representative images during (Figure 61A) and after (Figure 61B) the placement of a catheter for LEAD delivery into the portal vein. Figure 61C) Posterior view of an explanted rat liver following LEAD delivery into the right medial lobe; the orange arrow indicates the treated lobe. Figure 61D) IVIS image of the corresponding rat liver; the dashed circle highlights ICG fluorescence in the treated lobe. Figure 61E) Representative gross and IVIS images of transected rat liver lobes from control and LEAD embolized groups. Figure 61F) High-power microscopic images of liver histology sections stained with H&E, or immunostained with nivolumab, CK7, or CD31 from control or LEAD groups. Figure 61G) Quantifications of cell density, nivolumab distribution area, cholangiocyte count, bile duct count, and CD31+cell count in control and LEAD groups. Statistical differences were determined using unpaired t-tests. Data in all graphs are means ± SEM (n=3). ***P < 0.001, and ****P < 0.0001. Figures 62A – 62H. In vivo portal vein embolization, ablation, and drug delivery in a nonsurvival porcine model. Figure 62A) Representative image illustrating portal vein access under ultrasound (US) guidance. The line depicts the needle tract, while the arrow and asterisk indicate the needle tip and portal vein, respectively. Figures 62B – 62D) Representative digitally subtracted fluoroscopy images displaying pre-embolization portal vein angiography (Figure 62B), LEAD delivery in the portal vein branch supplying the left lateral lobe (Figure 62C), and post-embolization angiography using a 5F catheter (Figure 62D). The dashed lines outline the borders of the embolized porcine liver lobe, with the arrow in the post-embolization fluoroscopy image indicating successful embolization of the portal vein branches. Figure 62E) Gross image of the explanted porcine liver following LEAD delivery into the left lateral lobe. The arrows denote the embolized (LEAD) lobe and control lobe. Figure 62F) Representative cross- sectional gross and IVIS images of LEAD embolized porcine liver lobes illustrating uniform ablation and ICG distribution compared to control. Figure 62G) High-power microscopic images of porcine liver histology sections stained with H&E, or immunostained for nivolumab, CK7, or CD31 from control and LEAD groups. Figure 62H) Quantifications of cell density, nivolumab distribution, cholangiocyte count, bile duct count, and CD31+cell count in control and LEAD groups. Scale bar = 75 µm. Statistical differences were determined using unpaired t-tests. Data in all graphs are means ± SEM (n=7). ****P < 0.0001. Figures 63A – 63H. In vivo portal vein embolization, ablation, and drug delivery in a survival porcine model. Figure 63A) Representative image illustrating portal vein access under US guidance; the green dashed line depicts the needle tract, while the white arrow and white asterisk indicate the needle tip and portal vein, respectively. Figures 63B – 63D) Representative digital subtracted fluoroscopy images of pre-embolization portal vein angiography (Figure 63B), portal vein embolization with LEAD (Figure 63C), and angiography at 1 minute (1m) and 7 days (7d) after post-embolization (Figure 63D) using a 5F catheter. Dashed lines outline the borders of the embolized porcine liver lobe with arrows in the post- embolization fluoroscopy images showing the embolized portal vein. Figure 63E) Gross image of the explanted porcine liver following LEAD delivery into the left lateral lobe. The arrows denote the embolized (LEAD) lobe and control lobe. Figure 63F) Representative cross-sectional gross and IVIS images of LEAD embolized porcine liver lobes illustrating uniform ablation and ICG distribution compared to control at 7 days after embolization. Figure 63G) High-power microscopic images of porcine liver histology sections stained with H&E, or immunostained for nivolumab, CK7, or CD31 from control and LEAD groups. Figure 63H) Quantifications of cell density, nivolumab distribution, cholangiocyte count, bile duct count, and CD31+cell count in control and LEAD groups. Scale bar = 150 µm. Statistical differences were determined using unpaired t-tests. Data in all graphs are means ± SEM (n=5). ***P < 0.001, and ****P < 0.0001. Figures 64A – 64G. In vivo tumor ablation and drug delivery in a mouse ectopic cholangiocarcinoma tumor model. Figure 64A) Illustration outlining the construction of the murine model of cholangiocarcinoma and US-guided treatment with LEAD. Figure 64B) Gross appearance of the ectopic tumor on the back of a mouse. Figure 64C) LEAD injection into the tumor guided by US imaging. Figure 64D) Representative cross-sectional gross and IVIS images of explanted tumors from the control and LEAD groups. Positive signal in the cross-sectional IVIS image of the LEAD group indicates uniform ablation and ICG distribution. Scale bar = 5 mm. Figure 64E) Representative images of H&E and nivolumab-stained histology sections of tumors from the control and LEAD groups. Scale bar = 5 mm. Black boxes denote areas displayed in (Figure 64F). Figure 64F) Representative images showcasing high-power microscopic images of H&E and nivolumab-stained areas of control and LEAD groups. Scale bar = 75 µm. Figure 64G) Quantifications of cell density and nivolumab distribution in control and LEAD groups. Statistical differences were calculated using unpaired t-tests. Data in all graphs represent means ± SEM (n=5). ***P < 0.001, and ****P < 0.0001. Figures 65A – 65B. Doxorubicin diffusion after solubilizing in ILs. Figure 65A) Fluorescent image illustrating variable doxorubicin (Dox) diffusion in tissue-like matrices following solubilization in IL3:1, IL1:1, and IL1:3. Figure 65B) Graph illustrating the area of Dox diffusion in IL3:1, IL1:1, and IL1:3. Statistical differences were calculated using ANOVA and Tukey’s post-hoc tests. Data in all graphs are the means ± SEM (n ≥ 6). **P < 0.01, ***P < 0.001, and ****P < 0.0001. Figures 66A – 66B. Indocyanine green diffusion after solubilizing in IL3:1. Figure 66A) Near- infrared fluorescent images illustrating concentration-dependent indocyanine green (ICG) radial diffusion in a agarose matrix following solubilization in varying concentrations of IL3:1. Figure 66B) Graph illustrating the area of ICG diffusion in 10%, 30%, 50%, and 70% IL3:1 solutions. Statistical differences were calculated using ANOVA and Tukey’s post-hoc tests. Data in all graphs are the means ± SEM (n ≥ 3). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. Figures 67A – 67H. Assessing fractional viability in response to cell treatment with ILs. Dose- response plots demonstrating the fractional viability and IC50 values of SNU478 (Figures 67A and 67D), SB1 (Figures 67B, 67E, and 67G), HUCCT1 (Figures 67C and 67F), and HUVEC (Figure 67H) cells 24 hours post-treatment with serially diluted ILs, highlighting pronounced cytotoxicity at low concentrations across all tested cells. IC50values were extrapolated from the sigmoidal fitted curve of the viability values specific to each cell line corresponding to the log10 concentrations of the ILs. Data in all graphs are the means ± SEM (n ≥ 8). Figures 68A – 68D. Assessment of synergistic cytotoxicity potential of combined treatment with IL3:1 and cisplatin in cancer cells. Figure 68A) Fractional viability plot illustrating the response of SNU478 cells to varying concentrations (0-200 µM) of cisplatin. Figure 68B) A matrix displaying the viability response values of SNU478 cells at 24 hours post combination treatment with the indicated cisplatin concentrations (0-12.5 µM) and IL3:1 concentration (0.156%-1.25%). Figures 68C – 68D) 3D surface plot (Figure 68C) and synergy score matrix (Figure 68D) generated using the Loewe / Bliss model with Combenefit, demonstrating the highest cytotoxic synergy score of 37±1 with a combination treatment of 0.156% IL3:1 and 3.125% cisplatin against SNU478 for 24 hours. Data represent replicates of three independent experiments (n = 3). Figures 69A – 69C. FTIR spectroscopy of IL3:1 (Figure 69A), geranic acid (Figure 69B), and choline bicarbonate (Figure 69C) in the spectral region between 500-4000 cm-1wavenumber shows highly intense IR peaks characteristic of the selected chemical bond in the fingerprint region of its respective spectrum. Figures 70A – 70B. Zeta potential (Figure 70A) and conductivity (Figure 70B) measurements of IL3:1. Graph depicting concentration-dependent change in zeta potential and conductivity of serially diluted IL3:1. Data are reported as mean ± s.e.m. and are based on a minimum of three independent replicates. Figures 71A – 71B. Assessment of ICG stability. Figure 71A) Near-infrared images of ICG in multiwells loaded with either ICG solubilized in water (control, top row) or in LEAD (bottom row), showing the time-dependent stability of ICG fluorescence in LEAD compared to the control after incubation at 37°C. Figure 71B) Quantitative analysis depicting the stability of ICG fluorescence over 7 days in LEAD compared to diminished fluorescent intensity in the control group within 2 days after incubation at 37°C. Data are reported as mean ± s.e.m. and are based on a minimum of three independent replicates. Figure 72. Antibacterial assay. Representative blood agar plates illustrating antibacterial effects of LEAD at varying concentrations against patient-derived Pseudomonas aeruginosa (P. aeruginosa), vancomycin-resistant Enterococcus (VRE), and methicillin-resistant Staphylococcus aureus (MRSA) bacterial strains. Numbers (n) in the figure indicate the LEAD dilution factor (2n). Figures 73A – 73E. Assessment of ablation efficacy and drug diffusion in ex vivo human renal cell carcinoma (RCC) tissue. Figure 73A) Representative IVIS fluorescent images of RCC tissue at 1- hour post-intratumoral injection of LEAD compared to control. Figure 73B) Representative histology sections stained with H&E or immunostained for nivolumab in control (top) and LEAD-injected (bottom) RCC tissue. Figures 73C – 73E) Graphs depicting fold change in average radiant efficiency (Figure 73C), cell density (Figure 73D), or nivolumab (Figure 73E) detection area directly measured on explanted RCC tissues or evaluated on histology sections from control or LEAD-injected RCC tissues. Data are reported as mean ± s.e.m. and are based on a minimum of 3 independent replicates. Figures 74A – 74D. Assessment of ablation efficacy and drug diffusion in ex vivo human lung cancer (HLC) tissue. Figure 74A) Representative near-infrared fluorescent images of HLC tissues and graph depicting a time-dependent increase in ICG fluorescence up to 4-hours post-intratumoral injection of LEAD compared to control. Figure 74B) Representative HLC histology sections stained with H&E or immunostained for nivolumab in control (top) and LEAD-injected (bottom). Figures 74C – 74D) Graphs depicting fold change in mean cell density (Figure 74C) or nivolumab detection area (Figure 74D) evaluated on histology sections from control or LEAD-injected HLC tissues. Figures 75A – 75D. Rat model of portal vein embolization. An illustrative summary of the rat portal vein embolization model using LEAD, depicting an exemplary procedure. The figure outlines a process used to assess the ablation efficacy and drug delivery potential of LEAD, including surgical exposure (Figure 75A) of the rat portal vein (Figure 75B), portal vein catheterization (Figure 75C), followed by LEAD injection using a 1F catheter to cause embolization (Figure 75D). Figures 76A – 76I. Renal artery embolization, ablation, and drug delivery in a nonsurvival porcine model. Figures 76A – 76B) Representative digitally subtracted angiography (DSA) through a 5F catheter positioned in the main renal artery illustrates patent renal artery branches at baseline (pre- injection) (Figure 76A) and complete renal artery occlusion after the injection of LEAD (post-injection) (Figure 76B). Figures 76C – 76D) Representative anterior and posterior gross and near-infrared fluorescent images of the explanted porcine kidney before (Figure 76C) and after (Figure 76D) bisection following renal artery embolization with LEAD. Figure 76E) Representative tiled H&E-stained histology section of kidney tissue post-embolization with LEAD. Figure 76F) Representative images of H&E- stained, nivolumab-, or HIF2α-immunostained histology sections corresponding to the numbered black boxes in Figure 76E, obtained at 1 hour after renal artery embolization with LEAD compared to control. Figure 76G) Graph depicting the frequency of embolization based on arterial branch diameter evaluated across the renal histology sections. Figures 76H – 76I) Graphs illustrating morphometric measurements of cell density (Figure 76H) or nivolumab detection area (Figure 76I), evaluated on stained histology sections from control or after renal artery embolization with LEAD. Statistical differences were calculated using unpaired t-test. Data in all graphs are the means ± SEM (n = 4). **P < 0.01, and ****P < 0.0001. Figures 77A – 77C. Porcine model of portal vein embolization. An illustrative summary of the porcine model of portal vein embolization used to assess the efficacy of embolization, ablation, and drug delivery potential of LEAD, including percutaneous access to the portal vein using ultrasound imaging guidance (Figure 77A), portal vein catheterization (Figure 77B), followed by LEAD injection into the portal vein using a 5F Yueh catheter to achieve embolization, ablation, and drug delivery (Figure 77C). Figures 78A – 78C. In vivo portal vein embolization in a nonsurvival pig model. Figure 78A) Representative gross image of explanted porcine liver following the delivery of 70% PBS + 20% IOH (v / v) + 10% Nivo + 0.25 mg / mL ICG solution into the left lateral lobe. Arrows denote the treatment lobe and the control lobe. Figure 78B) Representative gross and IVIS images of transected control and treated lobes. Figure 78C) Representative images of H&E-stained, nivolumab-immunostained, and CK7- immunostained histology sections from control and treatment group. (n = 3). Figures 79A – 79E. Porcine model of portal vein embolization with LEAD. An illustrative summary of the porcine survival portal vein embolization model used to assess the embolization and ablation efficacy and drug delivery potential of LEAD. Upon percutaneous access to the portal vein using ultrasound imaging guidance (Figure 79A), portal vein catheterization (Figure 79B), and subsequent LEAD injection into the portal vein using a 5F Yueh catheter was performed to achieve embolization, ablation, and drug delivery (Figure 79C). Following a 1-week survival period (Figure 79D), the pigs were euthanized (Figure 79E). Figures 80A – 80B. Histological evaluation of liver morphology after embolization with LEAD. Figure 80A) Representative tiled microscopic images of H&E-stained liver histology sections obtained from the control liver or at 7 days after percutaneous portal vein embolization with LEAD. Figure 80B) Representative high-power microscopic images corresponding to the numbered boxes in (Figure 80A), showing serially cut histology sections stained with H&E, Picrosirius Red, or immunostained for the marker of bile ducts, cytokeratin-7 (CK7), the marker of macrophage / monocyte, CD68, or the marker of acute inflammatory cells, MPO. Scale bar = 50 µm. (n=3). Figures 81A – 81C. In vitro analysis of porcine blood hemolysis after treatment with LEAD. Figure 81A) Representative images of vials containing whole porcine blood treated with various concentrations of LEAD showing concentration-dependent hemolysis. Figure 81B) Graph illustrating hemolysis rate in porcine blood treated with 0.1%, 1%, 2%, 5%, or 10% LEAD. Figure 81C) Representative porcine blood smears after treatment with various concentrations of LEAD. Statistical differences were calculated using ANOVA and Tukey’s post-hoc tests. Data in are the means ± SEM (n ≥ 3). ns: not significant, ***P < 0.001, and ****P < 0.0001. Figures 82A – 82F. In vitro analysis of complete porcine blood counts in response to treatment with LEAD. Graphs illustrating red blood cell (Figure 82A), white blood cell (Figure 82B), lymphocyte (Figure 82C), monocyte (Figure 82D), granulocyte (Figure 82E), and platelet (Figure 82F) counts in porcine blood samples after one-hour incubation with varying concentrations of LEAD at 37°C. Statistical differences were calculated using ANOVA and Tukey’s post-hoc tests. Data in are the means ± SEM (n ≥ 4). ns: not significant, *P < 0.05, ***P < 0.001, and ****P < 0.0001. Figure 83. In vitro analysis of porcine blood thrombogenicity in response to treatment with LEAD. Image showing citrated blood after being mixed with varying LEAD concentrations (0.1%, 1%, 2%, 5%, or 10%) and subsequently activated with CaCl2to induce coagulation. The mixture was incubated for 1, 3, 5, 7, 10, 20, and 30 minutes at room temperature inside a multiwell plate, demonstrating complete blood coagulation at 5 minutes in the control compared to delayed coagulation at 1% LEAD or higher (n ≥ 3). Figure 84. In vitro analysis of ACT of porcine pig after treatment LEAD. Graph illustrating ACT values for porcine blood samples treated with varying concentrations of LEAD (0%, 0.1%, 1%, 2%, 5%, and 10%) demonstrating delayed coagulation with 1% LEAD treatment or higher. Statistical differences were calculated using ANOVA and Tukey’s post-hoc tests. Data in are the means ± SEM (n ≥ 3). *P < 0.05, and ****P < 0.0001. Figures 85A – 85B. Hemorheology of activated porcine blood treated with LEAD. Figure 85A) Representative G′ flow curves obtained from activated porcine blood samples after treatment with varying concentrations of LEAD (0%, 0.1%, 1%, 2%, 5%, and 10%), measured by a rheometer over a 30-minute at 37°C. Figure 85B) Graph depicting a decrease in storage modulus of coagulated blood (G′) at 30 minutes after treatment with 0.1% LEAD or higher. Statistical differences were calculated using ANOVA and Tukey’s post-hoc tests. Data in are the means ± SEM (n ≥ 3). ns: not significant, and ****P < 0.0001. DETAILED DESCRIPTION The terminology used herein is for the purpose of describing particular cases only and is not intended to be limiting. The below terms are discussed to illustrate meanings of the terms as used in this specification, in addition to the understanding of these terms by those of skill in the art. As used herein and in the appended claims, the singular forms “a”, “an”, and “the” include plural referents unless the context clearly dictates otherwise. It is further noted that the claims can be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as “solely,” “only” and the like in connection with the recitation of claim elements, or use of a “negative” limitation. Certain ranges are presented herein with numerical values being preceded by the term “about.” The term “about” is used herein to provide literal support for the exact number that it precedes, as well as a number that is near to or approximately the number (±10%) that the term precedes. In determining whether a number is near to or approximately a specifically recited number, the near or approximating un-recited number may be a number which, in the context in which it is presented, provides the substantial equivalent of the specifically recited number. Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the methods and compositions described herein are. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also encompassed within the methods and compositions described herein, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the methods and compositions described herein. The terms “individual,” “patient,” or “subject” are used interchangeably. None of the terms require or are limited to situation characterized by the supervision (e.g., constant or intermittent) of a health care worker (e.g. a doctor, a registered nurse, a nurse practitioner, a physician’s assistant, an orderly, or a hospice worker). Further, these terms refer to human or animal subjects. “Treating” or “treatment” refers to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) a targeted pathologic condition or disorder. Those in need of treatment include those already with the disorder, as well as those prone to have the disorder, or those in whom the disorder is to be prevented. Unless otherwise indicated, the term “percent” refers to weigh percentage, i.e. percent by weigh or “wt%”. When the disclosure specifically refers to volume percentage, it mean percent by volume or “%v / v”. The terms “effective amount” or “therapeutically effective amount,” as used herein, refer to a sufficient amount of an agent or a compound being administered which will relieve to some extent one or more of the symptoms of the disease or condition being treated. The result can be reduction and / or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. For example, an “effective amount” for therapeutic uses is the amount of the composition including a compound as disclosed herein required to provide a clinically significant decrease in disease symptoms without undue adverse side effects. An appropriate “effective amount” in any individual case may be determined using techniques, such as a dose escalation study. The term “therapeutically effective amount” includes, for example, a prophylactically effective amount. An “effective amount” of a compound disclosed herein is an amount effective to achieve a desired pharmacologic effect or therapeutic improvement without undue adverse side effects. It is understood that “an effect amount” or “a therapeutically effective amount” can vary from subject to subject, due to variation in metabolism of the compound, age, weight, general condition of the subject, the condition being treated, the severity of the condition being treated, and the judgment of the prescribing physician. By way of example only, therapeutically effective amounts may be determined by routine experimentation, including but not limited to a dose escalation clinical trial. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the methods and compositions described herein belong. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the methods and compositions described herein, representative illustrative methods and materials are now described. This document provides methods and materials for occluding blood vessels within a mammal (e.g., a human), ablating tissue (e.g., cancer tissue) or cells (e.g., cancer cells) in an area proximal to the blood occlusion site, and / or delivering agents (e.g., anti-cancer agents) to localized areas within the mammal (e.g., a human). For example, this document provides a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents and methods for using such compositions to occlude blood vessels, to ablate tissue or cells, and / or to deliver agents to localized areas within a mammal (e.g., a human). In some cases, a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be administered to a lumen of a blood vessel within a mammal having cancer to ablate at least some cancer cells of a tumor (e.g., a solid tumor) within the mammal. For example, a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be administered to a lumen of a blood vessel within a mammal to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration such that the blood occlusion site can prevent blood supply from reaching a targeted tissue such as targeted tumor tissue. In some cases, a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be administered to a lumen of a blood vessel within a mammal having cancer to deliver agents to localized areas within a mammal (e.g., a human). For example, a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be administered to a lumen of a blood vessel within a mammal to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration such that at least some components of the composition (e.g., a mixture of geranate and choline) can exit the blood vessel in the area of the blood occlusion site, enter the surrounding or nearby tissue, and ablate at least some tissue (e.g., tumor tissue) or cells (e.g., cancer cells) in that area. For example, a composition comprising an ionic liquid of choline geranate, glycerol, and optionally other compounds or agents (e.g., an anti-cancer agent) can be administered to a lumen of a blood vessel within a mammal to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration such that at least some components of the composition (e.g., ionic liquid, glycerol, and / or the other compound or agent) can exit the blood vessel in the area of the blood occlusion site, enter the surrounding or nearby tissue, ablate at least some tissue (e.g., tumor tissue) or cells (e.g., cancer cells) in that area, and deliver the other compound or agent to that area in a targeted manner such that the delivered compound or agent is primarily detectable at the blood occlusion site and the surrounding or nearby tissue. In some cases, a composition provided herein including an ionic liquid of choline geranate, and glycerol, and optionally one or more other compounds or agents for targeted delivery can be sterile. For example, sterile forms of choline geranate, glycerol, and one or more other compounds or agents can be mixed together under sterile conditions to form a composition that is sterile. A composition provided herein (e.g., a composition comprising an ionic liquid of choline geranate, glycerol, and optionally other compounds or agents such as an anti-cancer agent) can have any appropriate molar ratio of geranate to choline. In some cases, a composition provided herein can have a molar ratio of from about 1:3 (geranate:choline) to about 1:6 (geranate:choline). For example, a composition provided herein can have a ratio of from about 1:3 to about 1:5.5, from about 1:3 to about 1:5, from about 1:3 to about 1:4.5, from about 1:3 to about 1:4, from about 1:3.5 to about 1:6, from about 1:3.5 to about 1:5, from about 1:3.5 to about 1:4, or from about 1:3.5 to about 1:4.5 of geranate to choline. In some cases, a composition provided herein can have a ratio of geranate to choline of 1:4. In some cases, a composition provided herein can have molar ratio of from about 6:1 (geranate:choline) to about 1:6 (geranate:choline). For example, a composition provided herein can have a ratio of from about 5:1 to about 1:5, from about 4:1 to about 1:4, from about 3:1 to about 1:2, from about 2:1 to about 1:2. In some cases, the composition have a molar ratio of geranate to choline of about 6:1, about 5.5:1, about 5:1, about 4.5:1, about 4:1, about 3.5:1, about 3:1, about 2.5:1, about 2:1, about 1.5:1, about 1:1, about 1:1.5, about 1:2, about 1:2.5, about 1:3, about 1:3.5, about 1:4, about 1:4.5, about 1:5, about 1:5.5, or about 1:6. In some cases, the geranate anion of a composition provided herein can be replaced with another anion. Examples of anionic that can replace the geranate anion of a composition provided herein include, without limitation, bistriflimide, oleate, hexanoate, dodecyldimethyl ammonia propane sulfonate, N- lauryl sarcosinate, geraniolate, tetrafluoroborate, hexafluorophosphate, methyl sulfate, octyle sulfate, acesulfame, halides, bis(trifluoromethylsulfonyl)amide, bis(trifluoromethyl)amide, dicyanamide, and trifluoromethanesulfonate. The geranate anion of a composition provided herein is an anion of geranic acid and not an anion of geranic acid ester. In some cases, the choline cation of a composition provided herein can be replaced with another compound. Examples of cationic compounds that can replace the choline of a composition provided herein include, without limitation, benzyl pyridinium, benzyl dimethyl dodecyl ammonium, phosphonium, tetraalkylphosphonium, benzethonium, imidazolium, pyridinium, piperidinium, quinolinium, morpholinium, quaternary phosphonium, and quaternary ammonium. A composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can include any appropriate amount of glycerol. In some cases, at least 30 percent (e.g., about 30%, about 32%, about 35%, about 38%, about 40%, about 42%, or more) of a composition provided herein can be glycerol. In some cases, the glycerol of a composition provided herein (e.g., a composition including geranate, choline, and glycerol such as an ionic liquid composition including geranate, choline, and glycerol, and optionally one or more other compounds or agents for targeted delivery) can be replaced with another ingredient. For example, an ingredient that can replace the glycerol of a composition provided herein can be a sugar such as a sugar alcohol, a synthetic sugar, or a natural sugar. Examples of ingredients that can replace the glycerol of a composition provided herein include, without limitation, dextrose, fructose, galactose, glucose, lactose, maltose, polysorbates, sucrose, xylose, erythritol, glycerol, hydrogenated starch hydrolysates, isomalt, lactitol, maltitol, mannitol, sorbitol, xylitol, agave, honey, and molasses. A composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be in the form of a solution (e.g., an aqueous solution) or a suspension. A composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can have any appropriate viscosity. In some cases, a composition provided herein can have a viscosity of from about 0.1 centipoise (cP) to about 1000 cP (e.g., from about 0.1 cP to about 800 cP, from about 0.1 cP to about 600 cP, from about 0.1 cP to about 500 cP, from about 0.1 cP to about 400 cP, from about 0.1 cP to about 300 cP, from about 0.1 cP to about 200 cP, from about 0.1 cP to about 100 cP, from about 0.1 cP to about 50 cP, from about 1 cP to about 1000 cP, from about 10 cP to about 1000 cP, from about 50 cP to about 1000 cP, from about 100 cP to about 1000 cP, from about 200 cP to about 1000 cP, from about 300 cP to about 1000 cP, from about 400 cP to about 1000 cP, from about 500 cP to about 1000 cP, from about 700 cP to about 1000 cP, from about 1 cP to about 750 cP, from about 5 cP to about 500 cP, from about 10 cP to about 250 cP, from about 15 cP to about 90 cP, from about 20 cP to about 50 cP, from about 1 cP to about 100 cP, from about 50 cP to about 150 cP, from about 100 cP to about 200 cP, from about 150 cP to about 250 cP, from about 200 cP to about 300 cP, from about 250 cP to about 350 cP, from about 300 cP to about 400 cP, from about 350 cP to about 450 cP, from about 400 cP to about 500 cP, from about 450 cP to about 550 cP, from about 500 cP to about 600 cP, from about 650 cP to about 750 cP, from about 700 cP to about 800 cP, or from about 750 cP to about 850 cP). In some cases, a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can include one or more substances that can alter the viscosity of the composition. Examples of substances that can be included in a composition provided herein to alter the viscosity of the composition include, without limitation, acacias, agars, alamic acids, alginic acids, aluminum monostearates, attapulgites (e.g., activated attapulgite such as colloidal activated attapulgite), bentonites (e.g., bentonite magma and purified bentonite), polymers of acrylic acid (e.g., Carbomers such as Carbomer 1342, Carbomer 910, Carbomer 934, Carbomer 934P, Carbomer 940, Carbomer 941, Carbomer copolymer, Carbomer homopolymer, and Carbomer Interpolymer), carboxymethylcellulose calciums, carboxymethylcellulose sodiums (e.g., carboxymethylcellulose sodium 12), carrageenans, celluloses (e.g., microcrystalline cellulose), dextrins, gelatins, gellan gums, guar gums, hydroxyethyl celluloses, hydroxypropyl celluloses, hypromellose xanthan gums, magnesium aluminum silicates, maltodextrins, methylcelluloses, pectins, polyethylene oxides, polyethylene glycols, polyvinyl alcohols, povidones, propylene glycol alginates, silicon dioxides (e.g., colloidal silicon dioxide), sodium alginates, starches (e.g., corn starch, potato starch, tapioca starch, and wheat starch), and tragacanths. In some cases, a composition provided herein including choline geranate, and glycerol can include one or more (e.g., one, two, three, four, or more) additional compounds or agents. In some cases, an additional compound or agent that can be included in a composition provided herein can be a biologically active compound. An additional compound or agent that can be included in a composition provided herein can be any appropriate type of molecule (e.g., polypeptides, nucleic acids, and small molecules). Examples of compounds and agents that can be included in a composition provided herein include, without limitation, imaging agents, and therapeutic agents. An additional compound or agent that can be included in a composition provided herein (e.g., a composition including geranate, choline, and glycerol such as an ionic liquid composition including geranate, choline, and glycerol) can be any appropriate size. For example, a compound or agent that can be included in a composition provided herein can have a molecular weight that is greater than about 7,500 Daltons (e.g., greater than about 10,000 Daltons, greater than about 15,000 Daltons, greater than about 25,000 Daltons, greater than about 50,000 Daltons, greater than about 100,000 Daltons, or greater than 125,000 Daltons). For example, a compound or agent that can be included in a composition provided herein can have a molecular weight that is less than about 750,000 Daltons, less than about 500,000 Daltons, less than about 250,000 Daltons, or less than about 200,000 Daltons. For example, a compound or agent that can be included in a composition provided herein can have a molecular weight that is from about 7,500 Daltons to about 750,000 Daltons (e.g., from about 7,500 Daltons to about 500,000 Daltons, from about 7,500 Daltons to about 250,000 Daltons, from about 7,500 Daltons to about 100,000 Daltons, from about 7,500 Daltons to about 75,000 Daltons, from about 7,500 Daltons to about 50,000 Daltons, from about 7,500 Daltons to about 30,000 Daltons, from about 7,500 Daltons to about 15,000 Daltons, from about 7,500 Daltons to about 10,000 Daltons, from about 10,000 Daltons to about 750,000 Daltons, from about 15,000 Daltons to about 750,000 Daltons, from about 25,000 Daltons to about 750,000 Daltons, from about 50,000 Daltons to about 750,000 Daltons, from about 75,000 Daltons to about 750,000 Daltons, from about 100,000 Daltons to about 750,000 Daltons, from about 250,000 Daltons to about 750,000 Daltons, from about 500,000 Daltons to about 750,000 Daltons, from about 600,000 Daltons to about 750,000 Daltons, from about 10,000 Daltons to about 500,000 Daltons, from about 25,000 Daltons to about 250,000 Daltons, from about 50,000 Daltons to about 200,000 Daltons, from about 75,000 Daltons to about 150,000 Daltons, from about 25,000 Daltons to about 50,000 Daltons, from about 50,000 Daltons to about 100,000 Daltons, from about 100,000 Daltons to about 150,000 Daltons, from about 150,000 Daltons to about 200,000 Daltons, from about 200,000 Daltons to about 250,000 Daltons, from about 250,000 Daltons to about 300,000 Daltons, from about 300,000 Daltons to about 350,000 Daltons, from about 350,000 Daltons to about 400,000 Daltons, from about 400,000 Daltons to about 450,000 Daltons, from about 450,000 Daltons to about 500,000 Daltons, from about 500,000 Daltons to about 550,000 Daltons, from about 550,000 Daltons to about 600,000 Daltons, from about 600,000 Daltons to about 650,000 Daltons, from about 650,000 Daltons to about 700,000 Daltons, or from about 700,000 Daltons to about 750,000 Daltons). In some cases, a composition provided herein including choline geranate, and glycerol can include one or more (e.g., one, two, three, four, or more) imaging agents. In some cases, an imaging agent included in a composition provided herein can be a radiodense imaging agent. In some cases, an imaging agent included in a composition provided herein can be an earth metal-based imaging agent. In some cases, an imaging agent included in a composition provided herein can be compatible with magnetic resonance imaging. In some cases, an imaging agent included in a composition provided herein can be compatible with nuclear imaging. In some cases, an imaging agent included in a composition provided herein can be compatible with ultrasound imaging. In some cases, an imaging agent included in a composition provided herein can be compatible with fluorescent imaging. Examples of imaging agents that can be included in a composition provided herein include, without limitation, blue fluorescent proteins (BFPs), green fluorescent proteins (GFPs), red fluorescent proteins (RFPs), indocyanine green, ExiTron™, Lipiodol®, iohexol, tantelum (e.g., tantalum nanoparticles and tantalum microparticles), gold nanoparticles, gadolinium, indium111, iodine, and microbubbles. When a composition provided herein including choline geranate, and glycerol includes one or more (e.g., one, two, three, four, or more) imaging agents, the composition can include any appropriate amount of the imaging agent(s). For example, when a composition provided herein includes one or more imaging agents, at least 1 percent (e.g., 1 percent v / v) of the composition can be imaging agent(s). In some cases, when a composition provided herein includes one or more imaging agents, from about 1 percent (e.g., 1 percent v / v) to about 40 percent (e.g., 40 percent v / v) of the composition (e.g., from about 1 percent to about 35 percent, from about 1 percent to about 30 percent, from about 1 percent to about 25 percent, from about 1 percent to about 20 percent, from about 1 percent to about 15 percent, from about 1 percent to about 10 percent, from about 1 percent to about 5 percent, from about 5 percent to about 40 percent, from about 10 percent to about 40 percent, from about 15 percent to about 40 percent, from about 20 percent to about 40 percent, from about 25 percent to about 40 percent, from about 30 percent to about 40 percent, from about 35 percent to about 40 percent, from about 5 percent to about 35 percent, from about 10 percent to about 30 percent, from about 15 percent to about 25 percent, from about 5 percent to about 15 percent, from about 10 percent to about 20 percent, from about 20 percent to about 30 percent, or from about 25 percent to about 35 percent) can be imaging agent(s). In some cases, when a composition provided herein includes one or more imaging agents, from about 0.01 mg to about 500 mg (e.g., from about 0.01 mg to about 450 mg, from about 0.01 mg to about 400 mg, from about 0.01 mg to about 350 mg, from about 0.01 mg to about 300 mg, from about 0.01 mg to about 250 mg, from about 0.01 mg to about 200 mg, from about 0.01 mg to about 150 mg, from about 0.01 mg to about 100 mg, from about 0.01 mg to about 50 mg, from about 0.01 mg to about 10 mg, from about 0.05 mg to about 500 mg, from about 0.1 mg to about 500 mg, from about 1 mg to about 500 mg, from about 50 mg to about 500 mg, from about 100 mg to about 500 mg, from about 150 mg to about 500 mg, from about 200 mg to about 500 mg, from about 250 mg to about 500 mg, from about 300 mg to about 500 mg, from about 350 mg to about 500 mg, from about 400 mg to about 500 mg, from about 450 mg to about 500 mg, from about 0.05 mg to about 400 mg, from about 0.1 mg to about 300 mg, from about 0.5 mg to about 200 mg, from about 1 mg to about 100 mg, from about 0.05 mg to about 250 mg, from about 0.1 mg to about 10 mg, from about 0.1 mg to about 100 mg, from about 1 mg to about 100 mg, from about 10 mg to about 200 mg, from about 50 mg to about 300 mg, from about 100 mg to about 350 mg, or from about 150 mg to about 400 mg) per mL of the composition of the composition can be imaging agent(s). In some cases, a composition provided herein including choline geranate, and glycerol can include one or more (e.g., one, two, three, four, or more) therapeutic compounds or agents. In some cases, a therapeutic agent included in a composition provided herein can be a biologically active compound (e.g., a biologically active polypeptide). In some cases, a therapeutic agent (e.g., a therapeutic polypeptide) included in a composition provided herein can be an immunogenic polypeptide (e.g., a peptide that is immunogenic within a mammal such as a human). In some cases, a therapeutic agent (e.g., a therapeutic polypeptide) included in a composition provided herein can be an antigen-binding polypeptide (e.g., an antibody). Examples of antigen-binding polypeptides that can be included in a composition provided herein include, without limitation, nivolumab, ipilimumab, pembrolizumab, atezolizumab, durvalumab, cemiplimab, and avelumab. In some cases, a therapeutic agent included in a composition provided herein can have anti-cancer activity (e.g., within a mammal such as a human). When a composition provided herein including choline geranate, and glycerol includes one or more (e.g., one, two, three, four, or more) therapeutic compounds or agents, the composition can include any appropriate amount of the therapeutic compound(s) or agent(s). For example, when a composition provided herein includes one or more therapeutic agents, at least 0.01 percent of the composition can be therapeutic agent(s). In some cases, when a composition provided herein includes one or more therapeutic agents, from about 0.01 percent to about 10 percent of the composition can be therapeutic agent(s). In some cases, when a composition provided herein includes one or more therapeutic agents, from about 0.01 mg to about 500 mg (e.g., from about 0.01 mg to about 450 mg, from about 0.01 mg to about 400 mg, from about 0.01 mg to about 350 mg, from about 0.01 mg to about 300 mg, from about 0.01 mg to about 250 mg, from about 0.01 mg to about 200 mg, from about 0.01 mg to about 150 mg, from about 0.01 mg to about 100 mg, from about 0.01 mg to about 50 mg, from about 0.01 mg to about 10 mg, from about 0.05 mg to about 500 mg, from about 0.1 mg to about 500 mg, from about 1 mg to about 500 mg, from about 50 mg to about 500 mg, from about 100 mg to about 500 mg, from about 150 mg to about 500 mg, from about 200 mg to about 500 mg, from about 250 mg to about 500 mg, from about 300 mg to about 500 mg, from about 350 mg to about 500 mg, from about 400 mg to about 500 mg, from about 450 mg to about 500 mg, from about 0.05 mg to about 400 mg, from about 0.1 mg to about 300 mg, from about 0.5 mg to about 200 mg, from about 1 mg to about 50 mg, from about 0.05 mg to about 250 mg, from about 0.1 mg to about 10 mg, from about 0.1 mg to about 100 mg, from about 1 mg to about 100 mg, from about 10 mg to about 200 mg, from about 50 mg to about 300 mg, from about 100 mg to about 350 mg, or from about 150 mg to about 400 mg) per mL of the composition of the composition can be therapeutic agent(s). In some cases, a composition provided herein including choline geranate, and glycerol can include one or more nucleic acids. A nucleic acid included in a composition provided herein can be in any appropriate form (e.g., an expression vector, a viral vector, naked DNA, or naked RNA). A nucleic acid included in a composition provided herein can include single-stranded nucleic acid, double-stranded nucleic acid, or a combination thereof. In some cases, a nucleic acid included in a composition provided herein can encode an imaging agent described herein (e.g., a BFP). In some cases, a nucleic acid included in a composition provided herein can encode a therapeutic agent described herein. When a composition provided herein including choline geranate, and glycerol includes one or more (e.g., one, two, three, four, or more) nucleic acids, the composition can include any appropriate amount of the nucleic acid(s). In some cases, when a composition provided herein includes one or more nucleic acids, from about 0.01 mg to about 500 mg (e.g., from about 0.01 mg to about 450 mg, from about 0.01 mg to about 400 mg, from about 0.01 mg to about 350 mg, from about 0.01 mg to about 300 mg, from about 0.01 mg to about 250 mg, from about 0.01 mg to about 200 mg, from about 0.01 mg to about 150 mg, from about 0.01 mg to about 100 mg, from about 0.01 mg to about 50 mg, from about 0.01 mg to about 10 mg, from about 0.05 mg to about 500 mg, from about 0.1 mg to about 500 mg, from about 1 mg to about 500 mg, from about 50 mg to about 500 mg, from about 100 mg to about 500 mg, from about 150 mg to about 500 mg, from about 200 mg to about 500 mg, from about 250 mg to about 500 mg, from about 300 mg to about 500 mg, from about 350 mg to about 500 mg, from about 400 mg to about 500 mg, from about 450 mg to about 500 mg, from about 0.05 mg to about 400 mg, from about 0.1 mg to about 300 mg, from about 0.5 mg to about 200 mg, from about 1 mg to about 50 mg, from about 0.05 mg to about 250 mg, from about 0.1 mg to about 10 mg, from about 0.1 mg to about 100 mg, from about 1 mg to about 100 mg, from about 10 mg to about 200 mg, from about 50 mg to about 300 mg, from about 100 mg to about 350 mg, or from about 150 mg to about 400 mg) per mL of the composition of the composition can be nucleic acid(s). Any appropriate method can be used to obtain a composition provided herein (e.g., a composition including geranate, choline, and glycerol such as an ionic liquid composition including geranate, choline, and glycerol, and optionally one or more other compounds or agents for targeted delivery). In some cases, a composition provided herein can be obtained by combining geranic acid and choline bicarbonate in the presence of a solute. Examples of solutes that can be used to produce a composition provided herein include, without limitation, acetone, and ILE. In some cases, a composition provided herein can be obtained as described in Example 1. In some cases, a composition provided herein can be obtained as described elsewhere (see, e.g., U.S. Patent No.10,449,254 at, e.g., column 11, line 6 to column 14, line 66; WO 2022 / 036309 at, e.g., Example 1; and Albadawi et al., Sci. Transl. Med., 13:eabe3889 (2021) at, e.g., the paragraph bridging pages 8-9)). Also provided herein are methods for using a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents. In some cases, a composition provided herein can be used to treat a mammal (e.g., a human) having cancer. For example, a composition provided herein can be administered to a lumen of a blood vessel within a mammal (e.g., a human) having cancer to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration. In some cases, such a blood occlusion site can be formed to prevent blood supply from reaching a targeted tissue (e.g., targeted tumor tissue). An occlusion formed by a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be any appropriate size. In some cases, a composition provided herein can be administered to a lumen of a blood vessel within a mammal (e.g., a human) in need thereof (e.g., a mammal such as a human having cancer) to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration, and to reduce the blood supply that reaches a targeted tissue (e.g., targeted tumor tissue) by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent. In some cases, a composition provided herein can be administered to a lumen of a blood vessel within a mammal (e.g., a human) in need thereof (e.g., a mammal such as a human having cancer) to form a blood occlusion that completely or at least partially occludes the blood up to 20 cm downstream of the site of administration, and to reduce the blood supply that reaches a targeted tissue (e.g., targeted tumor tissue) by at least 2-fold (e.g., by 2-fold, 3-fold, 4-fold, 5-fold, or more). In some cases, a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be administered to a lumen of a blood vessel within a mammal (e.g., a human) having cancer to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration, and such that at least some components of the composition (e.g., the ionic liquid and / or additional agents) can exit the blood vessel in the area of the blood occlusion site, enter the surrounding or nearby tissue, and ablate at least some targeted tissue (e.g., targeted tumor tissue) or cells (e.g., cancer cells) in that area. For example, a composition provided herein can be administered to a lumen of a blood vessel within a mammal (e.g., a human) having cancer to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration, and such that at least some components of the composition (e.g., the ionic liquid and / or the other compound or agent) can exit the blood vessel in the area of the blood occlusion site, enter the surrounding or nearby tissue, and ablate, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent of the targeted tissue (e.g., targeted tumor tissue) or cells (e.g., cancer cells) in that area. For example, a composition provided herein can be administered to a lumen of a blood vessel within a (e.g., a human) in need thereof (e.g., a mammal such as a human having cancer) to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration, and such that at least some components of the composition (e.g., the ionic liquid and / or the other compound or agent) can exit the blood vessel in the area of the blood occlusion site, enter the surrounding or nearby tissue, and ablate targeted tissue (e.g., targeted tumor tissue) or cells (e.g., cancer cells) in that area to reduce the size of a cancer (e.g., to reduce the number of cancer cells in the mammal and / or to reduce the volume of one or more tumors in the mammal) within a mammal (e.g., a human). In some cases, a composition provided herein can be administered to a lumen of a blood vessel within a mammal (e.g., a human) having cancer to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration, and to reduce the size of the cancer by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent. In some cases, a composition provided herein can be administered to a lumen of a blood vessel within a mammal (e.g., a human) having cancer to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration, and to reduce the size of the cancer by at least 2-fold (e.g., by 2-fold, 3-fold, 4- fold, 5-fold, or more). A targeted tissue (e.g., targeted tumor tissue) or cells (e.g., cancer cells) that can be ablated when a mammal (e.g., a human) in need thereof (e.g., a mammal such as a human having cancer) is administered a composition provided herein (e.g., a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents) to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration, and when at least some components of a composition (e.g., the ionic liquid and / or the additional agent) exit the blood vessel in the area of the blood occlusion site, can be any appropriate distance from the occlusion site. For example, a composition provided herein can be administered to a lumen of a blood vessel within a mammal (e.g., a human) having cancer to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration, such that at least some components of the composition (e.g., the ionic liquid and / or additional agent) can exit the blood vessel in the area of the blood occlusion site, enter the surrounding or nearby tissue, to ablate at least some targeted tissue (e.g., targeted tumor tissue) or cells (e.g., cancer cells) that is from about up to about 20 cm from the occlusion site. A targeted tissue (e.g., targeted tumor tissue) or cells (e.g., cancer cells) that can be ablated when a mammal (e.g., a human) in need thereof (e.g., a mammal such as a human having cancer) is administered a composition provided herein (e.g., a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents) to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration, and when at least some components of a composition (e.g., the ionic liquid and / or additional agents) exit the blood vessel in the area of the blood occlusion site, can be any appropriate tissue. For example, a composition provided herein can be administered to a lumen of a blood vessel within a mammal (e.g., a human) having cancer to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration, such that at least some components of the composition (e.g., the ionic liquid and / or additional agent) can exit the blood vessel in the area of the blood occlusion site, enter the surrounding or nearby tissue, to ablate liver tissue, and / or kidney tissue. In some cases, a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be used to treat a mammal (e.g., a human) having a disease or disorder other than cancer that could benefit from the ablation of at least a portion of a tissue within the mammal. For example, a composition provided herein can be administered to a lumen of a blood vessel within a mammal (e.g., a human) having a disease or disorder associated with fat accumulation to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration, such that at least some components of the composition (e.g., the ionic liquid and / or additional agent) can exit the blood vessel in the area of the blood occlusion site, enter the surrounding or nearby tissue, to ablate fat tissue in that area. For example, a composition provided herein can be administered to a lumen of a blood vessel within a mammal (e.g., a human) having a heart disease or disorder to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration, such that at least some components of the composition (e.g., the ionic liquid and / or additional agent) can exit the blood vessel in the area of the blood occlusion site, enter the surrounding or nearby tissue, to ablate one or more blood clots in that area. For example, a composition provided herein can be administered to a lumen of a blood vessel within a mammal (e.g., a human) having an infection (e.g., a bacterial infection or a viral infection) to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration, such that at least some components of the composition (e.g., the ionic liquid and / or additional agent) can exit the blood vessel in the area of the blood occlusion site, enter the surrounding or nearby tissue, to infected tissue in that area. When a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents is administered to a lumen of a blood vessel within a mammal (e.g., a human) having an infection (e.g., a bacterial infection or a viral infection) to form a blood occlusion that completely or at least partially occludes the blood vessel such that at least a portion of an infected tissue within the mammal is ablated, the mammal can have any type of infection. In some cases, a composition provided herein can be administered to a mammal having a bacterial infection. In some cases, a composition provided herein can be administered to a mammal having a viral infection. When a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents is administered to a lumen of a blood vessel within a mammal (e.g., a human) having an infection (e.g., a bacterial infection or a viral infection) to form a blood occlusion that completely or at least partially occludes the blood vessel such that at least a portion of an infected tissue within the mammal is ablated, the infection can be caused by any type of pathogen. Examples of pathogens that can cause an infection in a mammal and that could benefit from the ablation of at least a portion of an infected tissue within the mammal include, without limitation, E. coli, S. aureus (e.g., MRSA), Enterococci (e.g., VRE), Enterobacteriaceae (e.g., CRE), C. difficile, C. auris (e.g., multidrug-resistant C. auris). In some cases, when a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents includes one or more other compounds or agents for targeted delivery, the composition can be administered to a lumen of a blood vessel within a mammal (e.g., a human) in need thereof (e.g., a mammal such as a human having cancer) to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration, and such that at least some components of the composition (e.g., the ionic liquid and / or additional compound / agent) can exit the blood vessel in the area of the blood occlusion site, enter the surrounding or nearby tissue to deliver the other compound(s) or agent(s) to that area in a targeted manner. For example, a composition including geranate, choline, and glycerol, and one or more other compounds or agents for targeted delivery can be administered to a lumen of a blood vessel within a mammal (e.g., a human) having cancer to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration, and such that at least some components of the composition (e.g., the ionic liquid and / or additional compound / agent) exit the blood vessel in the area of the blood occlusion site, enter the surrounding or nearby tissue, and deliver the other compound(s) or agent(s) to that area such that the delivered compound or agent is primarily detectable at the blood occlusion site and the surrounding or nearby tissue. In some cases, administering a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be effective to deliver at least a portion of the other compound(s) or agent(s) to a tissue (e.g., a targeted tissue) within a mammal. For example, a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be administered to a lumen of a blood vessel within a mammal (e.g., a human) in need thereof (e.g., a mammal such as a human having cancer) to form a blood occlusion within the vessel, and can deliver at least a portion of the compound(s) or agent(s) to a tissue (e.g., a targeted tissue) located within 20 cm of the blood occlusion site. In some cases, a composition including geranate, choline, and glycerol, and one or more other compounds or agents for targeted delivery can be administered to a lumen of a blood vessel within a mammal (e.g., a human) to form a blood occlusion within the vessel, and can deliver at least a portion of the compound to a tissue (e.g., a targeted tissue) located within 20 cm of the blood occlusion site and outside of the blood vessel. In some cases, a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be administered to a lumen of a blood vessel within a mammal (e.g., a human) to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration, and such that at least some components of the composition (e.g., the ionic liquid and / or additional compound / agent) exit the blood vessel in the area of the blood occlusion site, enter the surrounding or nearby tissue, and deliver at least a portion of the compound(s) or agent(s) to a tissue (e.g., a targeted tissue) located outside of the blood vessel such that the tissue located outside of the blood vessel contains more of the compound. For example, a composition including geranate, choline, and glycerol, and one or more other compounds or agents for targeted delivery can be administered to a lumen of a blood vessel within a mammal (e.g., a human) to form a blood occlusion that completely or at least partially occludes the blood vessel up to 20 cm downstream of the site of administration, and such that at least some components of the composition (e.g., the ionic liquid and / or additional compound / agent) exit the blood vessel in the area of the blood occlusion site, enter the surrounding or nearby tissue, to deliver at least a portion of the compound(s) or agent(s) to a tissue (e.g., a targeted tissue) located outside of the blood vessel such that the tissue located outside of the blood vessel contains more of the compound than another tissue (e.g., a non-targeted tissue) away from the blood occlusion site. A composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be used to deliver any appropriate amount (e.g., any appropriate dose) of one or more compounds or agents for targeted delivery (e.g., one or more anti-cancer compounds) to at least a portion of a targeted tissue (e.g., targeted tumor tissue) within a mammal. A composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be administered to (e.g., can be administered into the lumen of a blood vessel within) any appropriate mammal. Examples of mammals that can be administered a composition provided herein include, without limitation, humans, non-human primates such as monkeys, horses, bovine species, porcine species, dogs, cats, horses, cows, pigs, sheep, mice, rabbit, and rats. For example, a composition provided herein can be administered to a human. In some cases, a composition provided herein can be administered to a human having cancer to treat the human. When treating a mammal (e.g., a human) having cancer as described herein (e.g., by administering a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents), the cancer can be any type of cancer. In some cases, a cancer to be treated as described herein can include one or more solid tumors. A tumor to be treated as described herein can be a benign tumor or a malignant tumor. For example, a cancer to be treated as described herein can include one or more fat laden solid tumors. In some cases, a cancer to be treated as described herein can be a blood cancer. In some cases, a cancer to be treated as described herein can be a primary cancer. In some cases, a cancer to be treated as described herein can be a metastatic cancer. In some cases, a cancer to be treated as described herein can be a cancer that has escaped and / or has been non- responsive to chemotherapy (e.g., a chemoresistant cancer). Examples of cancers that can be treated as described herein (e.g., by administering a composition including geranate, choline, and glycerol such as an ionic liquid composition including geranate, choline, and glycerol, and optionally one or more other compounds or agents for targeted delivery) include, without limitation, liver cancers (e.g., HCCs), bile duct cancers (e.g., cholangiocarcinoma), pancreatic cancers (e.g., pancreatic adenocarcinomas), colorectal cancers (e.g., colorectal cancer liver metastasis (CRCLM)), renal cancers, ovarian cancers, breast cancers, prostate cancers, colon cancers, bladder cancers, lung cancers, thyroid cancers, melanomas, brain cancers, stomach cancers, cervical cancers, uterine cancers, skin cancers, synovial cancers, appendiceal cancers, adrenal cancers, sarcomas, and lymphomas. In some cases, methods of treating a mammal having cancer described herein (e.g., by administering a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents) also can include identifying the mammal as having cancer. Examples of methods for identifying a mammal as having cancer include, without limitation, physical examination, laboratory tests (e.g., blood and / or urine), biopsy, imaging tests (e.g., X-ray, PET / CT, MRI, and / or ultrasound), nuclear medicine scans (e.g., bone scans), endoscopy, and / or genetic tests. In some cases, methods for using a composition provided herein (e.g., a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents) can include delivering the composition to (e.g., directly into the lumen of) one or more blood vessels of a mammal (e.g., a human). A composition provided herein can be administered to a lumen of any type of blood vessel (e.g., an artery, a vein, or a capillary) within a mammal (e.g., a human). In some cases, a composition provided herein can be administered to a lumen of a blood vessel that feeds (e.g., provides blood supply to) a tumor (e.g., a solid tumor). In some cases, a composition provided herein can be administered to a lumen of a blood vessel at a location that is within 20 cm of a tumor (e.g., a solid tumor). A composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be administered to a lumen of a blood vessel within any appropriate tissue (e.g., a target tissue) within a mammal (e.g., a human). Examples of tissues that include one or more blood vessels and can be targeted by administering a composition provided herein include, without limitation, kidney tissues, liver tissues, brain tissues, prostate tissues, pancreatic tissues, and breast tissues. In some cases, a tissue that can include one or more blood vessels to which a composition provided herein can be administered to a lumen of can include one or more cancer cells. For example, a tissue that can include one or more blood vessels to which a composition provided herein can be administered to a lumen of can include one or more kidney cancer cells, one or more liver cancer cells, one or more brain cancer cells, one or more prostate cancer cells, one or more pancreatic cancer cells, one or more breast cancer cells, one or more lung cancer cells, one or more colon cancer cells, and / or one or more bladder cancer cells. A composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be administered to a mammal (e.g., a human) by any appropriate route. In some cases, a composition provided herein can be administered to a mammal (e.g., a human) by injection (e.g., percutaneous injection) into a lumen of a blood vessel such as an intravascular injection or an intra-articular injection). For example, a composition provided herein can be administered to a lumen of a blood vessel within a mammal (e.g., a human) by percutaneous injection directly into the lumen of the blood vessel. In some cases, a composition provided herein can be administered to a mammal (e.g., a human) without the need for any anesthesia (e.g., without the need for general anesthesia). In some cases, a composition provided herein can be administered to a mammal (e.g., human) using a guided injection (e.g., using ultrasound guidance). In some cases, when a composition provided herein is administered to a mammal (e.g., a human) by a percutaneous injection into a lumen of a blood vessel, a single injection can be used to administer the composition. In some cases, when a composition provided herein is administered to a mammal (e.g., a human) by a percutaneous injection into a lumen of a blood vessel, two or more (e.g., two, three, four, five, or more) injections can be used to administer the composition. A composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be administered to a mammal (e.g., a human) in any appropriate amount (e.g., any appropriate dose). In some cases, an effective amount of a composition provided herein can be any amount that can completely or at least partially occlude a blood vessel within a mammal (e.g., a human) as described herein without producing significant toxicity to the mammal. The effective amount can remain constant or can be adjusted as a sliding scale or variable dose depending on the mammal’s response to treatment. Various factors can influence the actual effective amount used for a particular application. For example, the frequency of administration, duration of treatment, use of multiple treatment agents, route of administration, and / or severity of the condition being treated may require an increase or decrease in the actual effective amount administered. A composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be administered to a lumen of a blood vessel within a mammal (e.g., a human) at any appropriate frequency. The frequency of administration can be any frequency that can completely or at least partially occlude a blood vessel within a mammal without producing significant toxicity to the mammal. For example, the frequency of administration can be from about from about once a week to about once a month, from about twice a month to about once a month, or from about once a month to about once every three months. The frequency of administration can remain constant or can be variable during the duration of treatment. As with the effective amount, various factors can influence the actual frequency of administration used for a particular application. For example, the effective amount, duration of treatment, use of multiple treatment agents, and / or route of administration may require an increase or decrease in administration frequency. A composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be administered to a lumen of a blood vessel within a mammal (e.g., a human) for any appropriate duration. An effective duration for administering a composition provided herein can be any duration that can completely or at least partially occlude of a blood vessel within a mammal without producing significant toxicity to the mammal. For example, the effective duration can vary from several weeks to several months, from several months to several years, or from several years to a lifetime. Multiple factors can influence the actual effective duration used for a particular treatment. For example, an effective duration can vary with the frequency of administration, effective amount, use of multiple treatment agents, and / or route of administration. In some cases, a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be administered to mammal (e.g., a human such as a human having cancer) to induce an inflammatory reaction within an ablation zone created by the composition. For example, a composition provided herein can be administered to a mammal (e.g., a human) to recruit T-cells (e.g., activated T-cells) to an ablation zone created by the composition. In some cases, a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents can be administered to mammal (e.g., a human such as a human having cancer) to facilitate entry of one or more T cells (e.g., activated T cells) into a tumor (e.g., to increase the amount of one or more T cells in the tumor) within the mammal. For example, a composition provided herein can be administered to a mammal (e.g., a human) in need thereof (e.g., a human having cancer) as described herein to increase the amount of one or more T cells in a tumor within the mammal by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent. Examples of T cells that can be increased in a tumor following administration of a composition provided herein include, without limitation, CD4+T cells, CD8+T cells, and natural killer T cells. In some cases when a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents is used to treat a mammal (e.g., a human) having cancer, the composition provided herein can be the sole active agent used to treat the cancer. For example, a composition including geranate, choline, and glycerol can be the sole active agent used to treat a mammal (e.g., a human) having cancer. For example, a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents (e.g. anti-cancer agents) can be the sole active agent used to treat a mammal (e.g., a human) having cancer. In some cases when a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents is used to treat a mammal (e.g., a human) having cancer, the composition provided herein can include one or more (e.g., one, two, three, four, five or more) additional therapeutic agents used to treat the cancer. In some cases, a therapeutic agent used to treat cancer can be a chemotherapy agent. In some cases, a therapeutic agent used to treat cancer can be a radioactive agent. In some cases, a therapeutic agent used to treat cancer can be an immunotherapy agent (e.g., an immune checkpoint inhibitor such as anti-PD-1 antibodies and / or anti-PD-L1 antibodies). In some case, a therapeutic agent used to treat cancer can be a stimulator of interferon (IFN) gene (STING) agonist. Examples of therapeutic agents used to treat a cancer that can be administered to a mammal having cancer together with a composition provided herein (e.g., a composition including geranate, choline, and glycerol such as an ionic liquid composition including geranate, choline, and glycerol, and optionally one or more other compounds or agents for targeted delivery) include, without limitation, doxorubicin, cisplatin, paclitaxel, olaparib, everolimus, mitomycin, radioactive isotopes (e.g., yttrium Y- 90, lutetium-177, actinium, fluorine-18, gallium-67, krypton-81m, rubidium-82, nitrogen-13, technetium- 99m, indium-111, iodine-123, xenon-133, and thallium-201), atezolizumab, bevacizumab, cabozantinib- s-malate, ramucirumab, pembrolizumab, lenvatinib mesylate, sorafenib tosylate, nivolumab, pemigatinib, pembrolizumab, ramucirumab, regorafenib, and abemaciclib. In some cases, the one or more additional therapeutic agents can be administered together with the composition provided herein (e.g., a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents). In some cases, the one or more additional therapeutic agents can be administered independent of the composition provided herein (e.g., a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents). When the one or more additional therapeutic agents are administered independent of the composition provided herein (e.g., a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents), the composition provided herein can be administered first, and the one or more additional therapeutic agents administered second, or vice versa. In some cases, methods for treating a mammal (e.g., a human) having cancer as described herein (e.g., by administering a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents) also can include subjecting the mammal to one or more (e.g., one, two, three, four, five or more) additional treatments (e.g., therapeutic interventions) that are effective to treat cancer. Examples of additional treatments that can be used as described herein to treat cancer include, without limitation, radiation therapy, surgery, percutaneous tumor ablation, transcatheter embolization, and cancer immunotherapy. In some cases, the one or more additional treatments that are effective to treat cancer can be performed at the same time as the administration of the composition provided herein (e.g., a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents). In some cases, the one or more additional treatments that are effective to treat cancer can be performed before and / or after the administration of the composition provided herein (e.g., a composition provided herein including choline geranate, and glycerol, and optionally one or more other compounds or agents). The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims. NON-LIMITING EXAMPLES Example 1 Liquid embolic for catheter-directed embolization, tissue ablation, and drug delivery Materials and methods Preparation and characterization of ILE in different ratios ILE was prepared from a mixture of geranic acid and choline bicarbonate at predetermined ratios as described elsewhere (Kim et al., Nat. Nanotechnol., 5:465-472 (2010)). Briefly, neat geranic acid (Sigma-Aldrich, St. Louis, MO) was purified five times via recrystallization method at -80°C in acetone. Purified geranic acid was then added with choline bicarbonate (Sigma-Aldrich) and stirred at ambient temperature until the CO2byproduct was no longer detected. Subsequently, residual H2O was removed by rotary evaporator (R-300, Buchi, New Castle, DE) at 60°C for 1 hour. The synthesized ILE was characterized using attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy (Lumos II with Alpha II add-on, Bruker, Kontich, Belgium). Rheological study All rheological measurements of samples were performed using a rheometer (MCR 302, Anton Paar, Torrance, CA) at 37°C. For rheological measurements, a sandblasted 25-mm aluminum shaft and aluminum plate were used, keeping the gap in between at 1 mm. Flow curve measurements of all samples were performed at a shear rate range from 100to 103s-1. During the measurements, a water trap was used to prevent samples from drying out. Quantification of viscosity was calculated based on the values from the 5001 / s shear rate. All measurements were performed at least 3 times. Injection force measurements The injectability of all samples were determined using a mechanical tester equipped with a 100N load cell (Instron 5942, Instron, Norwood, MA). Briefly, all material was loaded in 3 cc syringes (Medallion, Merit Medical, South Jordan, UT) and injected through a 110 cm 2.8 F microcatheter (PROGREAT®, Terumo Interventional Systems, Somerset, NJ) at a constant flow rate of 1 mL min-1. The injection force was recorded using the Bluehill version 3 software (Instron). Agar diffusion assay Tissue-mimicking matrix was created to assess the diffusion of small molecules loaded in ILEs. Agarose (11685660001, Roche, Mannheim, Germany) was measured and dissolved in pH 7.41x phosphate buffered saline (PBS) (Life Technologies, Bleiswijk, Netherlands) to the final concentration of 2%. The 3D-printed well-casting inserts (5 mm diameter) were placed in 6-well plates (CELLTREAT®Scientific Products, Pepperell, MA) and 11 mL of warm 2% agarose solution was poured into each well and left to cure in a sterile hood. Once cured, inserts were carefully removed and 250 µL of free dyes and ILEs incorporated with equal amounts of dyes were transferred into the center reservoir to investigate the diffusion. Indocyanine green (ICG) (USP, Rockville, MD) and doxorubicin (DOX) (Pfizer, New York, NY) were used at 0.25 and 1 mg / mL, respectively. The samples were incubated in a 37°C humidified chamber and at predetermined time points (0, 4, and 24 hours), the distribution of fluorescence radiating from the dyes was imaged using the IVIS®spectrum in vivo imaging system (PerkinElmer Inc., Waltham, MA) at 740 / 850 nm (ICG) and 460 / 560 nm (DOX) wavelengths. The diffusion coefficient of each ILE sample was calculated at 24 hour timepoint with the following formula: Cell culture HepG2 hepatocellular carcinoma cell line (CRL10741, ATCC, Manassas, VA) was cultured using an Iscove’s Modified Dulbecco’s Medium (IMDM; HyClone, Logan, UT) supplemented with 10% heat-inactivated bovine serum albumin (BSA; HyClone), 100 IU penicillin and 10 μg / mL streptomycin (Thermo Fisher Scientific, Waltham, MA), and 0.1 mM non-essential amino acids (Lonza, Walkersville, MD). Cells were cultured in a 37°C, 5% CO2humidified chamber and handled aseptically. TRANSWELL®diffusion assay In a 12-well permeable TRANSWELL®culture plate with 400 nm pore-sized polycarbonate membrane (230615, CELLTREAT®), 5 × 105cells / well of HepG2 cells were seeded in the lower compartment and incubated for 16 hours at 37°C, 5% CO2humidified chamber. Upon incubation, HepG2 cells were washed and replenished with a fresh growth medium.500 µL of samples (1:2 or 1:3 or 1:4 ILE+ICG) were transferred into the inserts and incubated for 24 hours (n= 3). After incubation, inserts were removed and ICG that has been diffused to the lower chamber through the pores was quantified using a microplate reader using 780 / 820 nm wavelengths (SpectraMax iD5, Molecular Devices, San Jose, CA). The viability of HepG2 cells was assessed using WST-1 cell assay kit (Cayman, Ann Arbor, MI) according to the manufacturer’s instructions. In vitro fractional viability assay 3 × 104cells / well of HepG2 were seeded in 96-well tissue culture plates (229196, CELLTREAT®) and incubated overnight in a 37°C, 5% CO2humidified chamber. Followed by incubation, cells were washed with a pre-warmed Dulbecco’s modified phosphate buffer (DPBS; Sigma- Aldrich) and treated with a fresh IMDM growth medium contained with serially 2-fold diluted samples (Maximum concentration: 25%). The samples were incubated for 24 hours followed by three repeats of washing with DPBS to remove dead cells. After the washing step, each well was added with 100 µL of growth medium and 10 µL of WST-1 reagent and incubated for 1.5 hours. The absorbance of samples was measured using a microplate reader (Molecular Devices) at 450 nm and the percentage of viable cells in each treated well was calculated relative to the control. The dose-response factional viability plot and corresponding IC50values were analyzed using Prism Software (GraphPad, San Diego, CA). Optimization of ILE To increase the viscosity of ILEs, different amount of glycerol (Sigma-Aldrich) ranging from 0 ~ 40% (v / v) was added. The viscosity of the resulting mixture at 37°C was measured with a rheometer (Instron). For real-time visualization during the embolization procedure, different amount of FDA- approved contrast agent iohexol (Omnipaque™, GE Healthcare Systems, Chicago, IL) was incorporated and examined using fluoroscopy (OEC Elite C-Arm, GE Healthcare Systems) and micro-CT scanner (Skyscan 1276, Bruker). The quantification of fluoroscopic intensities and radiodensities of different concentrations of IOH was quantified using ImageJ (National Institutes of Health, Bethesda, MD) and CTAn (Bruker) software. Next, the stability of nivolumab (Bristol Myers Squibb, Princeton, NJ) in ILE was investigated.10 mg / mL stock of nivolumab was diluted to 1 mg / mL in ILE and ILE+glycerol samples and incubated at 37°C. At predetermined time points up to day 28, the samples were evaluated by running sodium sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under non-denaturing conditions. Each sample was mixed with Laemmli protein sample buffer (1610747, Bio-Rad, Hercules, CA), loaded (5 µg / lane) onto stain-free gels (4568084, Bio-Rad), and electrophoresed at 100V for 70 minutes. The samples in the gel were transferred to a 0.2 µm polyvinylidene fluoride (PVDF) membrane (Bio-Rad) using a trans-blot turbo transfer system (Bio-Rad). Following a successful transfer, the samples in the membrane were visualized using a gel imaging system (Gel Dox XR+, Bio-Rad). Zeta potential measurements The zeta potential of ILE, ILE+glycerol, and ILE+glycerol+IOH+ICG+nivolumab samples (ILE12, 13, and 14) were loaded in capillary cells (DTS1070, Malvern Panalytical Ltd., Malvern, United Kingdom) and analyzed using a Zetasizer ultra (Malvern Panalytical Ltd.) at ambient temperature. The data was collected and processed with ZS Xplorer software (Malvern Panalytical Ltd.). In vitro ICG degradation and fluorescence intensity over time ICG dissolved in an aqueous solution (control) and other ILE samples with various formulations were loaded into black solid 96-well plates and visualized every 24 hours for 28 days using IVIS®. The radiance efficiency of ICG was quantified with Living Image®software (PerkinElmer Inc.). The change in radiance efficiency values were calculated relative to the values of day 0 as a control. In vitro drug release test The release profiles of nivolumab (146 kDa) and BSA (67 kDa) from ILE14 were examined using the Float-A-Lyzer®G2 dialysis device with 300 kDa-sized pores (Repligen Corporation, Rancho Dominguez, CA).2 mL of ILE(1:4)+glycerol+IOH+nivolumab and ILE(1:4)+glycerol+IOH+BSA were transferred into the dialysis tube and incubated at 37°C while being immersed in 6 mL of pH 7.41x PBS (Life Technologies). At predetermined time points, 25 µL of 1x PBS was taken out and the amount of protein was quantified using a Pierce™BCA protein assay kit (Thermo Fisher Scientific) according to the manufacturer’s instructions. Renal embolization in a porcine model All animal studies were performed according to animal protocols approved by the Mayo Clinic Institutional Animal Care and Use Committee (IACUC). Briefly, Yorkshire / Landrace pigs (Premier BioSource, Ramona, CA) weighing 50~55 kg were acclimated for 7 days under the supervision of a veterinarian. On the day of the procedure, anesthesia of pigs was induced by intramuscular injection of 5 mg / kg tiletamine-zolazepam (Telazol®, Zoetis, Parsippany-Troy Hills, NJ), 2 mg / mL xylazine, and 0.02 mg / kg glycopyrrolate, and was maintained with 1.5~2% of isoflurane inhalation. The iliac artery was percutaneously accessed under ultrasound guidance (Butterfly iQ+, Butterfly Network Inc., Guilford, CT) and a Bentson guidewire (Cook Medical, IN, USA) was placed.150 U / kg of heparin (Mylan, Canonsburg, PA) was intravenously administered to prevent coagulation during the procedure. Real-time DSA was used to navigate a 2.8 F microcatheter (Terumo Interventional Systems) and positioned inside the targeted renal artery branch. ILE (1:4), glycerol, ILE12, 13, 14, and beads (Bead Block™, Boston Scientific, Marlborough, MA) were injected into the targeted blood vessels under real-time fluoroscopy. After embolization, DSA was repeated to confirm vessel occlusion. The animals were euthanized at 1 hour post-embolization for the nonsurvival study and at 7 days post-embolization for the survival study. Blood samples were collected before embolization, after embolization, and at the terminal time-point for analysis. At necropsy, all kidneys and other major organs (heart, lung, liver, muscle, and spleen) were resected for analysis. Images of intact and bisected kidneys were taken immediately after the organ collection. Subsequently, ex vivo fluorescence imaging of cross-sectioned whole kidneys was performed using IVIS®and radiance efficiency was calculated using Living Image®software. Histological and Immunohistochemical (IHC) analysis Tissues were fixed in 10% buffered formalin (Fisher Scientific, Waltham, MA) for at least 7 days then processed for paraffin-embedding. All tissue sections were sliced in 5 µm sections and hydrated by serially immersing in xylene (twice, Fisher Scientific), 100% (three times), 95%, 80, 70, and 50% ethanol (Fisher Scientific) and distilled water for hematoxylin and eosin (H&E, Thermo Fisher Scientific) staining. For immunostaining, hydrated slides were subsequently immersed in 10 mM sodium citrate buffer and boiled to 95°C for 30 minutes followed by 30 minutes of cooling down in a water bath for antigen retrieval. After washing the slides in distilled water for 5 minutes, endogenous peroxidase quenching was performed with 3% hydrogen peroxide (Sigma-Aldrich) in 60% methanol (Fisher Scientific) for 30 minutes at ambient temperature. The slides were then washed with 0.05% PBS-T for 5 minutes and blocked using 5% goat serum (50197Z, Thermo Fisher Scientific) in 1x PBS for 1 hour to prevent non-specific binding. After the blocking step, cleaved caspase-3 (ab13847, 1:250, Abcam, Cambridge, MA), CD31 (ab182981, 1:200, Abcam), HIF-2α (ab109616, 1:1000, Abcam), PCNA (NB500-106, 1:1000, Novus Biologicals, Centennial, CO) primary antibodies diluted in 5% goat serum were added to each slide and incubated overnight at 4°C in a humidified chamber. The following day, the slides were washed three times with 0.05% PBS-T for 5 minutes each and horseradish peroxidase (HRP)- conjugated goat anti-rabbit IgG H&L (ab97051, 1:500, Abcam) was added to the tissues and incubated at room temperature for 1 hour. The samples were then washed three times with 0.05% PBS-T and developed using 3,3'-Diaminobenzidine (DAB) substrate (Vector Laboratories, Newark, CA) for 1 to 10 minutes. Once the color development is visible, the slides were rinsed in distilled water to halt further color development, counterstained in hematoxylin for 5 seconds, and mounted using a mounting medium (Richard-Allan Scientific, Fisher Scientific). For nivolumab detection in the porcine tissue, peroxidase- quenched slides were blocked using an avidin-biotin blocking buffer (004303, Invitrogen, Waltham, MA) for 1 hour at room temperature. After, a biotin-conjugated rabbit anti-human IgG4 secondary antibody (ab238617, 5 µg / mL, Abcam) diluted in a universal blocking reagent (FP1020, Akoya Biosciences, Marlborough, MA) was added to the slides and incubated for 1 hour at room temperature. For detecting nivolumab in the rabbit tissue, peroxidase-quenched slides were blocked in a universal blocking reagent overnight at 4°C in a humidified chamber. After, a biotin-conjugated rabbit anti-human IgG4 secondary antibody (1 µg / mL) was blocked in 5% rabbit serum for 1 hour. The blocked secondary antibody was directly added to the slides and incubated for 1 hour at room temperature. Streptavidin-HRP solution (51- 75477E, BD Biosciences, Franklin Lakes, NJ) was later applied to the slides and incubated for another 1 hour at room temperature. DAB staining, counterstaining, and mounting were performed. Endothelial cell counting was performed using Qupath software (University of Edinburgh) and IHC staining quantification was performed using ImageJ software. ILE (1:4) and ILE14 in blood For ACT measurements, anti-coagulated citrated blood (Innovative Research Inc. Novi, MI) was mixed with ILE (1:4) or ILE14 to the final concentrations of 0, 0.1, 1, 2, and 10% (v / v). Next, 0.2 M CaCl2(30:1, v / v, Sigma-Aldrich) was added to the samples and gently mixed to induce coagulation. The samples were instantly loaded in the Celite ACT cartridge (Abbott laboratories, Chicago, IL) and inserted into the i-STAT handheld critical blood analyzer (Abbott laboratories) for ACT analysis. The sample preparation for the hemorheology study was performed in the same preparation as in the ACT samples and flow curves were recorded as described in the “rheological study” section. The hemocompatibility of ILE (1:4) and ILE14 was evaluated via hemolysis assay. Briefly, anti- coagulated citrated blood was centrifuged at 2,000 rpm for 10 minutes and serum was aspirated. The remaining red blood cell (RBC) layer was resuspended in 150 mM sodium chloride to the original volume and further diluted to 5% (v / v) in 1x PBS. The prepared RBC samples were mixed with either ILE (1:4) or ILE14 to the final concentrations of 0, 0.1, 1, 2, 10% (v / v) and ammonium-chloride- potassium (ACK) lysing buffer (KD Medical, Columbia, MD) were used to prepare positive control. All samples were incubated at 37°C for 1 hour and centrifuged at 1,500 rcf for 5 minutes.150 µL of supernatant was transferred to a 96-well plate and the absorbance was measured at 450 nm. The hemolysis rate was calculated with the following formula: To assess the oxidation of blood mixed with different concentrations of embolic agents, the samples were prepared in the same way as the hemolysis assay except for the final centrifugation step. The samples were transferred to a 96-well plate and the absorbance spectrum (600~1,000 nm) was measured using a microplate reader. To investigate the fate of the innate and recruited immune cells in the presence of ILE, the blood sample was mixed with either ILE (1:4), glycerol or ILE14 to the final concentrations of 0.1, 1, 2 and 10% (v / v). The mixture was incubated at 37°C for 1 hour and analyzed using a veterinary hematology analyzer (HemaTrue®, Heska, Loveland, CO). Biocompatibility test Complete blood count, biochemical analysis and cytokine array of serum were performed using collected blood samples. CBC was analyzed with a veterinary hematology analyzer (Heska) and biochemical analysis was performed using a veterinary chemistry analyzer (DRI-CHEM 4000, Heska). Serum cytokine arrays were performed using the porcine multiplex cytokine array / chemokine array 13- plex (Eve Technologies, Calgary, CA). In vitro migration assay The migration assay was performed using a 35 mm µ-Dish with 2 well culture-insert (ibidi, Grafelfing, Germany).4 × 104HepG2 cells in 70 µL of supplemented IMDM growth medium were seeded in each insert well and further incubated until complete confluency. Once the cells have reached the confluency, the insert was carefully removed. The cells were washed twice with fresh 1x PBS and replenished with fresh supplemented IMDM growth medium containing sub-lethal doses (0.025, 0.05, 0.1% v / v) of ILE14 and incubated in a 37°C, 5% CO2humidified chamber. Every 24 hours, migration of the cells was observed using a cell imaging microscope (EVOS FL Auto 2, Thermo Fisher Scientific) until the area was completely closed in the control group. The experiments were done in triplicates. Embolization of the rabbit VX2 liver tumor arteries To generate a rabbit VX2 liver tumor model, an aliquot of a rabbit VX2 tumor homogenate that was kept in liquid nitrogen was thawed and resuspended in 1 mL of Dulbecco’s modified Eagle medium (DMEM, Thermo Fisher Scientific).1 mL of VX2 tumor homogenate suspension was injected into the calf muscle of a female New Zealand white rabbit (Charles River Laboratories, Wilmington, MA). After the injection, donor rabbits were left for 2-3 weeks for tumor growth. Once the tumor growth was confirmed with periodic ultrasound imaging, calf muscle tissue lesion with the tumor was harvested, and donor rabbits were euthanized. Harvested tumor tissue was immediately immersed in DMEM on ice and disassociated to about 1 mm3.2 pieces of tumor section were implanted in a liver medial segment of recipient rabbits using aseptic surgical techniques. After tumor implantation, a liver incision was compressed for at least 3 minutes using a gelatin sponge (Ethicon, Summerville, NJ) for hemostasis and wound closure. The tumor was left to grow until it reached 1-2 cm3based on ultrasound imaging. On the day of the procedure, the rabbits were anesthetized, the iliac artery was percutaneously accessed and a 0.014-inch REFLEX™steerable guidewire (Cordis, Miami Lakes, FL) was placed. Under the guidance of real-time DSA, a 2.4 F microcatheter with a 45° tip shape (Medtronic, Dublin, Ireland) was navigated to the hepatic artery. The microcatheter was further guided to the hepatic artery branch feeding the tumor- bearing liver segment and 1 cc of ILE14 was carefully injected into the targeted vessels for embolization. After embolization, DSA was repeated to confirm the vascular occlusion. At necropsy, the VX2 tumor bearing liver was exposed, photographed and liver tissues were harvested with other major organs (liver, heart, lung, kidney, and spleen) for fluorescence imaging and histological analysis. Ex vivo IVIS®imaging of intact and bisected tumor-bearing livers was performed and analyzed using Living Image®software. Ex vivo evaluation of diffusion and ablation in human cancer tissue Freshly explanted human kidney cancer tissues were injected with 20-30 µL of ILE14 into the central area of the tumor mass using a 30-gauge needle. The treated tissue samples were then placed in a humidified chamber and incubated in a complete RPMI 1640 medium at 37°C. After 1, 12, and 24 hours of injection, the samples were subjected to IVIS®imaging and then processed for histological analysis. Bacteria susceptibility test The bactericidal efficacy of ILE14 was tested using a green fluorescent protein (GFP) reporter- labeled Escherichia coli (E. coli) (25922GFP, ATCC) and patient blood culture isolates. GFP-E. coli was cultured overnight in a Luria-Bertani (LB) broth (Fisher Scientific) at 37°C / 230 rpm. Cultured GFP-E. coli was diluted in fresh LB broth to about 0.7 OD600and serially diluted ILE14 was added to the bacteria solution for further incubation. After 24 hours incubation at 37°C / 230 rpm, the samples were collected, and the fluorescence of bacteria suspension was evaluated using IVIS®and a microplate reader. For patient-derived pathogens, the stock was diluted to 0.5 McFarland standard absorbance (1.5 x 108CFU / mL) in 0.45% saline solution tubes (Remel, San Diego, CA). In a 96-well plate, 100 µL of bacteria solution and 100 µL of serially diluted (20to 210–fold) ILE14 were mixed (1:1 v / v). Following incubation at 37°C (C. difficile for 48 hours; the rest for 24 hours), the mixtures were directly plated on sheep blood agar plate (Remel) using cotton-tipped applicators (MEDLINE, Northfield, IL) and further incubated to assess bacterial susceptibility. Susceptible ILE14 concentrations exhibited no bacterial growth, intermediate ILE14 concentrations showed decreased bacterial colonies compared to the control, and resistant ILE14 concentrations showed comparable bacterial growth to the control. Results The molar ratio between the anionic (geranic acid; GA) and cationic (choline; C) monomers of the ionic liquid embolic (ILE) was varied to develop a formulation optimized for tissue ablation and drug delivery. Seven versions of the ILE were synthesized; these varied in the ratio of GA: C ranging from 4:1 to 1:4 (Figure 1A, Figure 7A, and 7B). To identify a formulation that would allow hand injectability for ease of use; the viscosity and the corresponding injection of force of each ILE were measured. The viscosity of the ILEs at 37°C and the force required to inject the ILE through a 2.8 French (F) (length: 110 cm) microcatheter indicated that 4:1, 1:2, 1:3, and 1:4 formulations to be the most suitable for injection using microcatheters (Figure 1B and 1C). Next, the diffusivity of the ILEs was tested using an agar-based diffusion assay inspired by an immunodiffusion assay (Restrepo et al., Appl Microbiol, 23:132-137 (1972)); fluorescent dyes indocyanine green (ICG; used as a drug surrogate) and doxorubicin (DOX), were used to visualize the diffusion capability of each ILE (Figure 1D and1E). At 24 hours, ILE ratios of 1:2, 1:3, and 1:4 demonstrated the highest level of circumferential diffusion compared to control samples. To show that the diffusion of the fluorescent dye in Figure 1D and 1E also represented the spreading of ILE, a functional diffusion assay was performed to measure the viability of hepatocellular carcinoma cells (HepG2). Using a TRANSWELL®plate with 400 nm-sized membrane pores (Figure 1F), which is similar to the size of pores in hyperpermeable vessels in solid tumors (Mperkris et al., P Natl Acad Sci USA, 114:1994-1999 (2017); Stylianopoulos et al., Proc Natl Acad Sci USA, 110:18632-18637 (2013)), ILE ratios 1:2, 1:3 and 1:4 demonstrated maximal diffusion across the TRANSWELL®into the cell culture chamber resulting in near zero viability of the HepG2 cells (Figure 1G and 1H). To determine the minimum amount of ILEs required for cell death, IC50values for each ILE were determined (Figure 1I). Fractional viability assays showed that all three candidate ILEs have potent cytotoxic effects with IC50values of 0.24 %, 0.39 %, and 0.54 % for the 1:2, 1:3 and 1:4 formulation, respectively (Figure 1I). The viability of HepG2 cells in Figure 1H and the IC50data of HepG2 cells in Figure 1I collectively indicated that at least 13% of the ILE from the TRANSWELL®likely diffused into the lower chamber within 24 hours. These data suggested that 1:2, 1:3, and 1:4 formulations of the ILEs have favorable viscosity for microcatheter injection and robust diffusion capability in agar and TRANSWELL®diffusion assays mimicking tissue matrix. To limit the effects of ILE to the arterial distribution, the viscosity of the ILE formulations was increased by adding glycerol, a biocompatible and viscous additive often used as a therapeutic preservative (Figure 9) (Takamura et al., J Petrol Sci Eng, 98:50-60 (2012)). Rheological measurements showed that the final concentration of 40 % (v / v) glycerol significantly increased the viscosity of the ILEs compared to controls (Figure 2A) without impacting their ability to diffuse (Figure 10A). Next, to achieve precise delivery of the ILE using clinical imaging tools such as fluoroscopy and computed tomography (CT), IOH, an FDA-approved aqueous contrast agent, was added to the ILE. The amount of IOH mixed with ILE was balanced by X-ray visibility, material properties, and cytotoxicity. The concentration-dependent increase in IOH corresponded to a proportional increase in radiodensity on fluoroscopic and CT images; 20% (v / v) of IOH was selected as an acceptable amount for visibility using X-ray imaging. Furthermore, the visibility of the ILEs with 20% IOH did not change when mixed with glycerol (Figure 2B and 2C). Next, the stability of nivolumab was tested, which is the most commonly used immunotherapy drug worldwide, when mixed with different ratios of ILE with or without glycerol. Results showed that nivolumab did not undergo degradation over a 24 hour incubation period when mixed with various ILE formulations (Figure 2D). To ensure the stability of nivolumab over a longer period, the analysis was continued up to 28 days at 37°C; no significant degradation of the protein was observed (Figure 11). To understand the charge distribution of the various ILE formulations (Table 1), zeta potential was measured. When glycerol was incorporated, the zeta potential shifted from positive to negative indicating a greater likelihood to diffuse rapidly throughout tissues (Figure 2E) (Kim et al., Nat Nanotechnol, 5:465-472 (2010)). Next, to examine the capability of the ILE formulation to preserve smaller molecules, near-infrared imaging agent ICG mixed with ILE was tested. Within 24 hours, the fluorescent signal from the free ICG was barely detectable and the signal was completely lost within 2 days (Mindt et al., Photoch Photobio Sci, 17:1189-1196 (2018)). However, when ICG was mixed with ILE with and without glycerol, the fluorescence signal remained significantly higher up to 28 days when compared to the control group indicating that ILE+glycerol has the capability to stabilize large proteins and small molecule structures (Figure 12). The release of nivolumab (146 kDa) from the ILE formulation to achieve drug delivery was examined. ILE mixed with nivolumab inside a dialysis tubing showed rapid release of nivolumab reaching nearly 90% within 24 hours. The release behavior of a smaller protein, bovine serum albumin (BSA; 67 kDa) was also examined showing a burst release reaching 67% within 24 hours and subsequent sustained release up to about 86% on day 5 (Figure 13). Viscosity measurements of the final ILE formulations containing ICG, IOH, and nivolumab demonstrated a range of 20-30 mPa*s, which is approximately 10-fold higher than the viscosity of whole blood at 37°C (Nader et al., Front Physiol, 10:1329 (2019)). The break-loose and injection force of ILEs also increased; however, the ILEsy were still easily injectable using a 2.8 F microcatheter (Figure 2G). Fractional viability assays of ILEs with ICG, IOH, and nivolumab showed a slight increase in IC50values compared to ILE alone, however, still highly lethal to the cancer cells (Figure 2H). Table 1. Composition of the three ILE formulations used for catheter-mediated embolization of the renal artery in swine. To assess ILE’s ability to embolize, ablate, and deliver drugs locally, a porcine renal embolization model was used, which has a hierarchical vascular network similar to tumors. Under real- time fluoroscopic imaging guidance, the lower pole of the kidney was embolized with either ILE(1:2)+glycerol+IOH+ICG+nivolumab (ILE12), ILE13, or ILE14, allowing the use of the upper pole of the same kidney as a negative control. As the ILEs exited the tip of the microcatheter, ILEs demonstrated excellent X-ray radiodensity reaching smaller distal arteries in the renal parenchyma and allowing real-time visibility, indicating the completion of embolization (Figure 14, Figure 3A, and 3B). These images also showed that the embolization of the lower pole arteries was instant; subsequent DSA imaging of the renal artery showed an absence of flow in the lower pole arteries that received ILEs (Figure 3B and Figure 15 – Figure 17). On necropsy, the kidneys visibly demonstrated the lower half of the kidney that received ILEs to appear darker in color, and on IVIS®imaging, ICG distribution showed that the darker areas on gross evaluation corresponded to ILE delivery (Figure 3C). On sagittal sections of the kidney, IVIS®images further indicated uniform detection of the ICG throughout the lower half of the kidney suggesting successful delivery into the renal parenchyma (Figure 3D, 3H, and 3I Figure 15 – Figure 17). ILE delivery into the kidneys demonstrated similar ICG detection levels and markedly higher ICG fluorescence in the lower pole when compared to the untreated upper pole (Figure 3H and 3I). On histology, lower pole arteries were embolized containing ILE and blood vessels also demonstrated no nuclei implying ablation compared to untreated upper pole arteries (Figure 3E). Immunohistochemical (IHC) staining of cleaved caspase-3 showed apoptosis throughout the lower pole of the ILE-treated kidneys suggesting that the ILE diffused trans-arterially into the renal parenchyma to achieve tissue ablation (Figure 3F and 3K). To further show that trans-arterial delivery occurred with ILE, in addition to uniform ICG detection and apoptosis, nivolumab was immunostained. Figure 3G and 3L showed that nivolumab could be trans-arterially delivered throughout the renal parenchyma; however, ILE14 demonstrated a greater level of delivery. To show that glycerol enhanced the delivery of the ILE and its components, separate porcine renal embolization experiments were performed with bare ILE (1:4) and glycerol alone. Injection of ILE into the lower pole of the kidneys led to instant but incomplete embolization and glycerol demonstrated immediate recanalization suggesting that they are not effective embolic agents alone. Both bare ILE (1:4) and glycerol alone also presented reduced levels of tissue ablation and drug delivery when compared to the ILE14 formulation (Figure 18 – Figure 21). Liquid embolic agents were injected into blood vessels leading to significant interactions with blood. To evaluate the effect of ILE14 on blood coagulation potential and hemocyte count, the ACT and hemorheology of various ILE14 concentrations and bare ILE (1:4) were tested. ILE14 up to 1% (v / v) and bare ILE at 0.1% (v / v) when mixed with blood demonstrated thrombosis similar to untreated blood. However, the blood remains anticoagulated at ILE14 concentrations greater than 1% and ILE alone greater than 0.1% and did not demonstrate the ability to thrombose (Figure 4A – 4D). Furthermore, hemolysis was minimal at concentrations up to 2%; however, lymphocytes demonstrated significant resistance at concentrations up to 10% (Figure 4E – 4G, and Figure 22). ILE14 and glycerol alone each demonstrated preservation of various hemocytes up to 10% concentration, possibly due to the preservative properties of glycerol (Figure 23) (Lee et al., J Appl Physiol, 100:615-622 (2006); Dervieux et al., J Biomed Opt, 25:105001-105001 (2020)). These results indicate that the proximal embolization observed in Figure 3 was unrelated to thrombosis but was instead related to the viscosity of the material, while downstream embolization may have been caused by thrombosis. In addition, these results showed that the cellular components of blood were minimally affected by ILE14 suggesting that the delivery of nivolumab to the ablated renal parenchyma can enhance the chances of achieving immunotherapy in solid tumors. To determine the durability of ILE14 embolization, nivolumab and ICG delivery and tissue ablation, survival experiments were performed and the results were compared to clinically used chemoembolization beads (Bead Block™) (Figure 24) (Chen et al., Front Chem, 7:408 (2019)). Beads were suspended in a mixture that contained an equal amount of nivolumab and ICG in ILE14. At baseline, both beads and ILE14 were able to achieve embolization (Figure 5A). However, on day 7, the upper pole of the kidneys that received bead embolization all demonstrated significant recanalization; the lower pole of the kidneys that received ILE14 demonstrated persistent embolization with no evidence of any recanalization implying that the vessel occlusions were permanent (Figure 5A and 5B). On gross examination of the kidneys, the upper pole that received beads showed heterogeneity with ischemic areas and normally perfused areas when compared to the contralateral normal kidneys. However, the lower pole of the kidneys that received ILE14 demonstrated significant ischemic injury with uniform atrophy and ablation of the renal parenchyma (Figure 5C). On IVIS®imaging, only the lower pole of the kidneys that received ILE14 showed ICG fluorescence suggesting successful small molecule delivery to the tissue parenchyma that did not wash away up to day 7 (Figure 5D, 5F, and Figure 25). On histology, marked differences were observed. Both the untreated and the bead-embolized kidneys showed similar cellularity with normal endothelial cell (CD31) immunostaining suggesting that tissue ablation was not observed in the sections evaluated (Figure 5E and Figure 26). Hypoxia-inducible factor-2α (HIF-2α) was detected at greater levels in the bead-embolized samples when compared to the controls suggesting a state of increased hypoxia from temporary embolization. ILE14-treated samples demonstrated occluded arteries with no signs of CD31 or HIF-2α immunostaining suggesting persistent embolization and ablation of the arterial wall (Figure 5E and 5G – 5I). Similarly, apoptosis in the bead-embolized kidneys was patchy and confined mostly to the glomeruli with no evidence of nivolumab detection. In the ILE14-treated samples, apoptosis was uniformly detected throughout the renal cortex with significant delivery of nivolumab. These results suggested that ILE14 achieved durable and permanent embolization and marked tissue ablation with significant drug delivery. ILE14 demonstrated no signs of toxicity when compared to untreated controls, immediately post- embolization, and to animals on day 7. Complete blood cell (CBC) counts, biochemical analysis of serum (Figure 27), serum cytokine array (Table 2), and histological evaluation of major organs (Figure 28) showed no signs of abnormality. Furthermore, an estimation of the potential pharmacodynamics of ILE14 based on the calculation of physiologic concentrations showed that the systemic amount of ILE will be too low to cause any detectable toxicity (Figure 29). Since tumor margins are often the sites of tumor recurrence, whether 5 to 25-fold dilution of the ILE14 from its IC50(0.57%) had an effect on the migration of HepG2 cells was examined. ILE14 as low as 0.025% (v / v) was able to suppress the migration of HepG2 (Figure 30). This suggested that while the local concentration of ILE14 may be under the threshold of cell death, ILE14 can still exert a therapeutic benefit by reducing cellular migration and potentially reduce recurrence. To determine whether ILE14 can embolize, ablate an arterial distribution, and deliver therapeutics to this ablated zone, the highly malignant rabbit VX2 orthotopic liver tumor model was used. Once the liver tumors reached a certain size based on ultrasound imaging (Figure 6A), a vascular embolization procedure was performed using the ILE14. Similar to clinical cases, a microcatheter was used in combination with a microwire to super- select the segmental artery that perfused the VX2 tumor. With the angiography image magnified to help visualize the material exiting the catheter tip, ILE14 was slowly injected using a 1 cc syringe, as would be done clinically, until stasis of flow was achieved. DSA imaging following ILE14 injection showed immediate embolization (Figure 6B, Figure 31A and 31B). At 1 hour post-embolization, gross and IVIS®images of the explanted liver were obtained, which showed preferential delivery and accumulation of the small molecule ICG into the VX2 liver tumor tissue (Figure 6C – 6F, Figure 31C and 31D). A histology analysis of the tumor was performed to determine whether ICG accumulation at the liver tumor also implied delivery of the ILE and nivolumab. On H&E and PCNA immunostained sections, control tumors showed cellularity with no evidence of nivolumab delivery. The ILE14 embolized tumors demonstrated marked tissue ablation and nivolumab delivery resulting from transarterial diffusion as evidenced by hypocellularity, significantly reduced PCNA staining, and a marked increase in nivolumab detection surrounding the embolized tumor arteries (Figure 6G – 6L, Figure 32, Figure 33). An off-target ILE14 injection demonstrated viable VX2 tumor and the absence of any nivolumab detection, indicated that therapeutic delivery and tissue ablation were specific to targeted ILE14 injection (Figure 34). Table 2: Summary of cytokine and chemokine levels in pig blood. Serum samples collected from pigs at baseline (before embolization) and at 7 days post-embolization (post-embo) showed no significant differences in cytokines and chemokine levels. Data are presented as mean ± SEM (n= 5). Statistical significance was determined using paired t-test. ns, not significant. To demonstrate whether ILE14 could diffuse and ablate human tumor matrix, freshly resected human renal cell carcinoma tumors were injected with ILE14 within one hour of resection. IVIS®imaging and quantitative analysis showed increased fluorescence radiance and diffusion area over 24 hours (Figure 35A – 35C) with histology showing a near-complete ablation of the tumor tissue, compared to the untreated samples (Figure 35D). Furthermore, nivolumab immunostaining of the tissues showed uniform distribution of nivolumab across large tissue area suggesting that ILE14 was also capable of delivering drugs throughout the human tumor matrix similar to the rabbit VX2 tumor (Figure 35E). To determine whether the unique tissue ablation capability of ILE14 extends to infections, susceptibility testing using antibiotic-resistant patient-derived pathogens was performed. ILE14 demonstrated major bactericidal effects on highly drug-resistant strains including, methicillin-resistant Staphylococcus aureus (MRSA; MIC: 6.25%), vancomycin-resistant Enterococcus faecium (VRE; MIC: 6.25%), and carbapenem-resistant Enterobacterales (CRE; MIC: 12.5%), Clostridioides difficile (C. difficile; MIC: 6.25%), and Candida auris (C. auris; MIC: 3.13%) (Figure 36). These results suggested that while currently used embolics may be prone to procedural infections (Guimaraes et al., Semin Intervent Radiol, 28:350-356 (2011)), ILE14 can achieve embolization, tissue ablation, and drug delivery, and, at the same time, protect the patient against abscess formation and potentially fatal sepsis even at very low concentrations. Example 2: Biocompatible liquid embolic for the treatment of microvascular hemorrhage Materials and methods Preparation and optimization of P-LE The ionic liquid was prepared by a metathesis reaction of geranic acid (Sigma-Aldrich, St. Louis, MO) and choline bicarbonate (Sigma-Aldrich). Initially, neat geranic acid was purified using the recrystallization method at a temperature of -80°C in acetone. Subsequently, the purified geranic acid was combined with choline bicarbonate at predefined ratios of 2:1, 1:1, or 1:2. The mixture was stirred at 25°C until the handheld carbon dioxide probe (GM70, VAISALA, Vantaa, Finland) indicated the absence of CO2byproduct. Following the completion of the reaction, any remaining H2O was removed using a rotary evaporator (R-300, Buchi, New Castle, DE) at 60°C for 1 hour. To enhance IL's viscosity and promote blood gelation, 100 kDa polyethylene glycol (PEG, Sigma-Aldrich) was added (100 mg / mL). To completely dissolve PEG in IL, neat PEG was added to IL and magnetically stirred at 60°C for 48 hours. Anti-coagulated blood (Innovative Research Inc., Novi, MI) was mixed with each IL or IL+PEG mixture (1:1 v / v). The hemorheology of mixture samples was measured using a rheometer (MCR 302, Anton Paar, Torrance, CA) at 37°C. A rheometer was outfitted with a sandblasted 25-mm aluminum shaft (Anton Paar) and aluminum plate (Anton Paar), maintaining a 1 mm gap in between. Storage modulus measurements were conducted on all samples for 30 minutes. Then, different amount of the FDA-approved contrast agent OMNIPAQUE™(IOH, GE Healthcare Systems, Chicago, IL) was incorporated, and the samples were subjected to fluoroscopy (OEC Elite C-Arm, GE Healthcare Systems) for real-time visualization during the embolization procedure. The intensity of the fluoroscopic signal of different concentrations of IOH was quantified using ImageJ (National Institutes of Health, Bethesda, MD). Upon finalizing the formulation of P-LE, hemorheology measurement was repeated to ensure that the addition of IOH did not hinder the ability of liquid embolic to gelate blood. For in vivo studies, ICG (United States Pharmacopeia, North Bethesda, MD) was incorporated into P-LE at a final concentration of 0.25 mg / mL to assess P-LE delivery post-embolization. Blood gelation assay using P-LE and clinically used coils Saline (Baxter Healthcare Corporation, Deerfield, IL), or P-LE was mixed with anti-coagulated blood (1:1 v / v) with or without an embolic coil (TORNADO® embolization microcoil, Cook Medical, Bloomington, IN) in a 12-well plate. The samples were incubated at 37°C and at predetermined time points, all wells were washed 3 times using saline to remove residues before imaging. Injection force measurements The injectability of P-LE was determined using a mechanical tester (Instron 5942, Instron, Norwood, MA). P-LE was loaded in 1 cc syringe (Medallion, Merit Medical, South Jordan, UT) and injected through a 100 cm 2.8 F microcatheter (PROGREAT®, Terumo Interventional Systems, Somerset, NJ) at a constant flow rate of 1 mL / minute. The injection force was recorded using the Bluehill version 3 software (Instron). Displacement pressure measurements The displacement pressure of the created ex vivo embolus was measured with a custom-built displacement pump set-up (WPI; pressure sensor from PASCO Capstone). Tested materials (IL, PEG, and P-LE) were mixed with blood, and 1 cc of the mixture was quickly placed at the center of the silicone tubing using a 2.8F microcatheter. After assembling the set-up, the infusion syringe pump (Kent Scientific, Torrington, CT) loaded with a 50 mL syringe (Henke Sass Wolf, Tuttlingen, Germany) filled with anti-coagulated blood was activated at a constant 30 mL / minute flow rate. All measurements were performed 3 times for quantitative analysis. Blood clotting index (BCI) assessments Anticoagulated blood was first placed in a 10 mL glass vial (Duran Wheaton Kimble, Rockwood, TN) and P-LE was slowly added (10% v / v) with or without the addition of CaCl2(30:1, v / v, Sigma- Aldrich). After gentle mixing, the samples were incubated at 37⁰C for 5 minutes. Saline was gently added to the samples without disrupting the formed clots. BCI was calculated as, BCI= 100%-(A450of the sample’s supernatant / A450of negative control) x 100%. Each measurement was repeated at least 5 times using citrated, heparinized (Innovative Research Inc.), and defibrinated (Innovative Research Inc.) blood for quantitative analysis. Activated clotting time (ACT) assessments using different blood components EDTA-blood (Innovative Research Inc.) was used to obtain each blood component. Platelet poor plasma was prepared by centrifugation at 3,000g for 10 minutes. Platelet rich plasma (supernatant) and red blood cells (cell pellets) were prepared by centrifugation at 2,000 rpm for 10 minutes. Prepared separate blood components were mixed with P-LE (10% v / v) with or without CaCl2and immediately loaded into the Celite ACT cartridge (Abbott Laboratories, Chicago, IL) and the i-STAT handheld blood analyzer (Abbott Laboratories) for the measurements. Femoral artery embolization in normal and heparinized rat model Sprague-Dawley rats with weights ranging from 300 to 400g were randomly divided for the femoral artery embolization procedure. The animals were anesthetized via inhalation of 2 L / minute of isoflurane (Piramal Critical Care, Mumbai, India) while resting on a warming pad. Positioned in a supine posture, the hair in the inguinal region was clipped and applied with povidone–iodine (Purdue Products, Stamford, CT). A 2.0 cm incision was created in parallel with the inguinal ligament to expose the femoral artery. A vascular sheet was removed, and the proximal FA was ligated using a 7-0 silk suture (Teleflex Medical, Plymouth, MN). Two ligatures were positioned distally on the FA for the injection of the embolic material to be tested. Within the space delineated by the two distal ligatures, a 30-gauge syringe was used for the intra-arterial injection of either PEG, IL, or P-LE. Following the injection, the proximal ligature was promptly released to allow blood flow into the injected materials. Five minutes after the removal of the proximal FA ligature, the distal ligatures were also removed to evaluate blood vessel occlusion by monitoring for any signs of breakthrough bleeding through the needle puncture site. For heparinized rats, a dosage of 250U / kg of heparin (Fresenius Kabi, Lake Zurich, IL) was administered 10 minutes prior to performing the embolization procedure. Hindlimb and FA of all rats were subjected to gross examination and LSCI (PeriScan PIM 3, Perimed Inc., Las Vegas, NV). At necropsy, the FA and surrounding tissues were harvested and examined under a micro-CT scanner (Skyscan 1276, Bruker, Kontich, Belgium) to visualize the distribution of injected materials. Renal embolization in non-survival and survival porcine model Renal artery embolization was performed as described elsewhere (Hu et al., Adv Mater, 34:e2106865 (2022); Albadawi et al., Adv Sci, 8:2003327 (2020)). Briefly, Yorkshire pigs (Premier BioSource, Ramona, CA) with a weight range of 50~60 kg were acclimated for 7 days under the supervision of a veterinarian. Before the procedure, pig anesthesia was initially induced through intramuscular injection of 5 mg / kg tiletamine-zolazepam (TELAZOL®, Zoetis, Parsippany-Troy Hills, NJ), 2 mg / mL xylazine, and 0.02 mg / kg glycopyrrolate, and then maintained with 2% of isoflurane inhalation. To access the renal artery, the iliac artery was initially accessed under the guidance of ultrasound imaging (Butterfly iQ+, Butterfly Network Inc., Guilford, CT). Subsequently, a guidewire (Cook Medical) was carefully placed. Real-time DSA was used to guide and accurately position a 2.8 F microcatheter (Terumo Interventional Systems, Somerset, NJ) within the targeted lower renal artery branch. P-LE was introduced into the lower lobe of the kidney under real-time fluoroscopic imaging. For survival study, clinically used 300 micron EMBOSPHERE®was injected into the upper renal lobe for comparison. After the P-LE injection, DSA was repeated to confirm the successful embolization of targeted renal artery branches. The pigs were humanely euthanized with EUTHASOL®(Virbac, Westlake, TX) one hour after embolization for the nonsurvival study and 14 days after embolization for the survival study, respectively. During necropsy, all kidneys were harvested and bisected for gross examination, as well as for IVIS®spectrum in vivo imaging system (PerkinElmer Inc., Waltham, MA) imaging. The IVIS imaging was taken using an ICG filter (740 / 850 nm) and quantified using Living Image®software (PerkinElmer). Renal embolization in a non-survival heparinized canine model Mongrel breed canines (Tri-Valley Resources, LLC, Spring Green, WI) with weights ranging from 35 to 50 kg were randomly divided for the embolization procedure (approval: A00007058-23). To sedate animals, an intramuscular injection of a cocktail comprising 0.2 mg / kg Midazolam (Avet Pharmaceutical, East Brunswick, NJ), 0.1 mg / kg Butorphanol (VCA Inc., Los Angeles, CA), and 5 mg / kg Propofol (Zoetic Inc., Richmond VA) was used. To induce heparinization in the canines, a dose of 250U / kg of heparin was administered 10 minutes before the procedure. The procedure for accessing the renal artery of the canines was performed identically to porcine models. Given the unique kidney structure in canines, the entire kidney was subjected to embolization. The canines were humanely euthanized one hour after the embolization, and all kidneys were collected for gross examination and IVIS imaging. Histological and immunohistochemical (IHC) analysis The tissues were fixed in 10% buffered formalin (Fisher Scientific, Waltham, MA) for a minimum of 7 days before being prepared for paraffin embedding. Tissue sections were sliced to a thickness of 5 µm and underwent a hydration process by sequentially immersing them in xylene (Fisher Scientific), followed by a series of ethanol (Fisher Scientific) solutions at concentrations of 100%, 95%, 80, 70, and 50%, and finally distilled water. For immunostaining of the slides, previously hydrated slides were placed into a 10mM sodium citrate buffer and subjected to boiling at 95°C for 30 minutes followed by 30 minutes of cooling down for antigen retrieval. The slides were washed with 3 changes of distilled water and background quenching was performed by immersing them in a solution of 3% hydrogen peroxide (Sigma-Aldrich) in 60% methanol (Fisher Scientific) for 30 minutes at room temperature. Subsequently, the slides were washed with 0.05% PBS-T for 5 minutes and then blocked for one hour using 5% goat serum (50197Z, Thermo Fisher Scientific). After the blocking step, CD31 (ab182981, 1:1000, Abcam), HIF-2α (ab109616, 1:1000, Abcam), MPO (ab208670, 1:500, Abcam) primary antibodies in 5% goat serum were pipetted onto each slide and incubated overnight at 4°C. On the following day, the slides were consecutively washed three times with 0.05% PBS-T. Horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG H&L (ab97051, Abcam) at a 1:500 dilution was then applied to the tissue sections and incubated at room temperature for one hour. Afterward, the samples were once again washed three times with 0.05% PBS-T. Subsequent 3,3'-Diaminobenzidine (DAB) substrate (Vector Laboratories, Newark, CA) incubation for up to 10 minutes was performed to develop the color. Once the desired color development was achieved, the slides were rinsed in distilled water to stop any further color development. The developed slides were briefly counterstained with hematoxylin for 5 seconds and mounted with a medium (Richard-Allan Scientific, Fisher Scientific). All quantitative analyses of the slides were performed using QuPath (University of Edinburgh) and ImageJ software. Toxicology studies The SpO2of survival study porcine subjects was measured at baseline, immediately after embolization, and 14 days after embolization. ACT, CBC, and serum biochemical analyses were performed using collected blood samples. ACT was analyzed using the Celite ACT cartridge (Abbott Laboratories) and the i-STAT handheld blood analyzer (Abbott Laboratories). CBC was analyzed using a veterinary hematology analyzer (Heska, Loveland, CO) and serum biochemical analysis was performed using a DRI-CHEM 4000 chemistry analyzer (Heska). Evaluation of P-LE in porcine microvascular hemorrhage models Prior to inducing acute hemorrhage, a 2.8F microcatheter was positioned at the desired location within the hepatic or splenic artery branches via percutaneous femoral artery ultrasound-guided access. Initially, DSA was conducted to observe the baseline vasculature by injecting IOH (350 mg / mL) through the catheter. To cause bleeding in the liver and stomach, Stiff™ guidewire (Boston Scientific, Marlborough, MA) was pushed through a microcatheter multiple times to induce vascular hemorrhage. To induce vascular hemorrhage in the renal tissue, under image guidance, an 18-gauge, 15 cm long needle was percutaneously inserted to injure the peripheral parenchyma of the renal cortex where the blood vessels are typically sub-mm. Angiography was performed to visualize contrast extravasation indicative of bleeding. Subsequently, the microcatheter was placed just proximal to the bleeding site, and P-LE was injected until stasis was observed. A final angiography was performed to ensure hemostasis was achieved. Bacteria susceptibility test The bacteria susceptibility test of P-LE was performed using multiple drug-resistant patient- derived isolated pathogens (MRSA, VRE, CRE, C. difficile, and C. auris). These microbes were first diluted in 0.45% saline (Remel, San Diego, CA) to 0.5 McFarland absorbance. After mixing 100 µL of each microbe with 100 µL of serially diluted P-LE, the mixture was incubated at 37°C. Aerobic microbes (MRSA, VRE, CRE, and C. auris) were incubated for 24 hour and anaerobic microbe (C. difficile) was incubated for 48 hours. Upon incubation, the mixes were plated directly onto sheep blood agar (Remel) using a cotton applicator (MEDLINE, Northfield, IL) without dilution. The plates were then further incubated at 37°C to evaluate susceptibility. Susceptible concentrations: no growth; Intermediate concentrations: reduced growth compared to control; Resistance concentrations: No growth change compared to control. Statistical Analysis Data are presented as mean ± SEM. Statistical analysis was performed by unpaired t-test for 2 groups or one-way ANOVA followed by Tukey’s comparison test for multiple groups unless otherwise specified. All analyses were performed using GraphPad Prism 10. All P values are 2-tailed, and P values less than or equal to 0.05 were considered statistically significant. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. NS, no significant. Results Microvascular hemorrhage can be seen in patients with chronic inflammation or infections, such as cystic fibrosis or tuberculosis, leading to hemoptysis; neoplastic conditions or minor trauma are other causes of microvascular hemorrhage, especially in patients who are on anticoagulants (Kleinbongard and Heusch, Nat Rev Cardiol, 19:265 (2022); de Vries et al., Virchows Arch, 460:629 (2012); Fukumura et al., Microcirculation, 17:206 (2010); Savioli et al., Medicines (Basel), 8 (2021); Maegele, Dtsch Arztebl Int, 116:799 (2019)). In these cases, the strategy for treatment often involved embolizing accessible upstream blood vessels. By occluding these larger, upstream vessels, flow to the hemorrhaging microvessels was interrupted, effectively treating the bleed. This approach was taken because directly targeting the numerous and tiny sub-millimeter vessels was not feasible (Figures 37A – 37C). This study introduced a novel liquid embolic material composed of PEG and a biocompatible IL, named P-LE. As illustrated in Figure 38A, this innovative material was specifically designed to reach and effectively embolize the distal microvasculature, a challenging target often inaccessible with current clinical tools. Characterization and optimization of P-LE The study investigated a novel IL formulation composed of choline (C) and geranic acid (G) at varying molar ratios (2:1, 1:1, 1:2) for its potential as a small vessel embolic agent. This IL was analyzed for its interaction with blood and its ability to occlude blood vessels measuring 30–50 microns. Rheology experiments revealed that increasing geranic acid's ratio led to a significant rise in the gelation rate (G'), implying its role in enhancing coagulation or interaction with blood cells (Figure 38B). Further testing with separated blood components showed that plasma, white blood cells, and platelets had minimal impact on gelation (Figures 38C and 38D). However, mixing IL with the red blood cell (RBC) fraction caused immediate gelation, suggesting that RBCs are required for effective embolization (Figure 38E). The study also considered variable hematocrit levels in patients, which may result from factors including bleeding, chemotherapy, or radiotherapy. It was found that higher RBC levels corresponded with increased G' levels, confirming the RBC dependency of the gelation process (Figures 38F and 38G). Additionally, the involvement of the coagulation cascade was examined using anticoagulated blood samples. The results indicated that the gelation of the blood-IL mixture occurred independently of the coagulation cascade, as anticoagulated blood did not inhibit the gelation process (Figures 38H – 38J). The ideal embolic agent should only embolize the target site upon release from the catheter, avoiding migration or fragmentation and preventing unintended embolization. To assess the stability of the IL-blood mixture, displacement pressure tests were conducted in a simulated blood flow system (Figure 39A). These tests revealed that the IL-blood mixture exhibited displacement forces comparable to blood clots formed by the addition of CaCl2, indicating a potential risk of fragmentation or migration of the IL-blood mixture (Figure 39B). To enhance the viscosity and stiffness of the IL-blood gel, high molecular weight PEG was integrated into the IL formulation. Various amounts of PEG were dissolved in IL to determine the appropriate PEG levels to include in the final formulation. The viscosity measurement of IL with PEG showed the highest viscosity and the fastest blood gelation when the PEG concentration reached 100 mg / mL. PEG concentrations higher than 100 mg / mL resulted in a precipitate, indicating that they exceeded the solubility threshold. This suggested that 100 mg / mL of PEG could be included in the formulation (Table 3) (Figure 44). This addition increased the viscosity of the IL, imparting a shear-thinning property beneficial for catheter injection (Figure 39C). The IL-PEG combination led to a quicker and more pronounced increase in modulus upon mixing with blood, compared to the IL alone (Figure 39D). Further investigation was conducted on the impact of this formulation on the ACT of anticoagulated blood, using different combinations of IL, PEG, and CaCl2. The results showed that while CaCl2induced thrombosis in anticoagulated citrated blood, lowering the ACT, PEG alone did not alter the ACT. However, any combination that included IL resulted in immediate thickening of the blood, underscoring IL's role in blood gelation. These outcomes highlight IL's effectiveness as a component in the formulation of the liquid embolic agent. (Figure 45). Table 3. Viscosity measurements of IL+PEG at different PEG concentrations. To assess delivery of the IL+PEG mixture using fluoroscopic imaging in clinical settings, IOH, an FDA-approved contrast agent, was incorporated into the mixture. Fluoroscopic evaluation indicated that the ideal concentration of IOH was 30% by volume, resulting in a new formulation termed P-LE (IL+PEG+30% IOH) (Figure 39E). Although the addition of 30% IOH slightly reduced the IL+PEG content in P-LE, the formulation maintained its gelling properties (Figure 39F). The injection force of P-LE was measured through a 2.8F microcatheter, confirming its suitability for hand-injection in endovascular procedures (Figure 39G). Further experiments focused on the speed of blood gelation with P-LE, a crucial aspect for successful vessel occlusion in medical applications. Tests with anticoagulated citrated blood demonstrated that P-LE induced instant gelling of the blood in comparison to the control and embolization coil group, indicating that P-LE is a potent embolic agent capable of functioning effectively even in patients with coagulopathic conditions, an outcome that holds considerable significance in clinical settings. Furthermore, when P-LE was injected within the coil mass of a failed coil embolization using a microcatheter, the combination resulted in the immediate gelation of blood. This finding is particularly significant for clinical scenarios where coil embolization fails in coagulopathic patients; however, with P-LE, these failed cases can be rescued (Figure 46). Ex vivo P-LE displacement test To evaluate the stability of a P-LE-mediated vascular occlusion against the forces exerted by systolic blood pressure, displacement pressure measurements were conducted. These measurements were consistent with the rheology findings shown in Figure 39D. When blood was combined with IL or CaCl2, the resulting blood clots exhibited displacement forces that indicated a potential risk for fragmentation. In contrast, blood mixed with PEG as part of the P-LE formulation exhibited significantly greater resistance. The measured displacement forces, in this case, were eight times higher than the normal systolic blood pressure, suggesting robust stability of the embolization in vivo (Figures 39H and 39I). P-LE embolization in heparinized rat femoral artery The durability and effectiveness of P-LE-mediated embolization in rat femoral arteries were investigated, focusing on the extent of distal small vessel embolization (Figure 47). Additionally, to demonstrate that P-LE embolization is independent of coagulation, experiments were conducted using heparinized rats. For control purposes, FA embolization with either PEG or IL alone was initially performed. Post-PEG injection into the FA, there was notable breakthrough bleeding from the puncture site, indicating the absence of vessel occlusion (Figure 40A). On histology, the FA was patent with no embolization and vessel wall ablation, supporting our in vitro findings that PEG alone is ineffective as an embolic agent (Figures 40B and 40C). Similarly, FA embolization with IL alone also resulted in breakthrough bleeding, suggesting incomplete embolization. Laser Speckle Contrast Imaging revealed minimal changes in arterial blood flow following IL injection, compared to baseline levels (Figure 40D). Hindlimb perfusion assessments indicated an insignificant change in perfusion rate after IL-only embolization, implying insufficient embolization (Figures 30E and 40F). High-resolution Micro-CT imaging demonstrated that embolization with IL alone was not sustained, as it was predominantly washed out from the FA and its side branch microvessels (Figure 40G). Histologically, the IL-injected FA also remained patent with vessel wall cellularity similar to the untreated control and to the PEG injection. CD31 immunostaining showed no significant difference between the control and IL-injected FA, also indicating the lack of vessel occlusion (Figures 40H and 40I). Collectively, these findings suggest that neither PEG nor IL alone can achieve complete embolization. When PEG and IL were combined in the P-LE formulation, embolization was successful without any breakthrough bleeding (Figure 40J). This was confirmed by hindlimb LSCI analysis demonstrating reduced blood perfusion in the embolized limb, suggesting on-target embolization was achieved by P-LE (Figures 40K and 40L). Following embolization, the FA and its downstream branches were imaged using micro-CT to visualize whether P-LE was able to reach distal microvessels. Analysis of micro-CT images showed that P-LE can reach microvessels smaller than 40 microns in diameter (Figure 40M). In contrast, OBSIDIO™ gel embolic was not able to reach sub-mm downstream vessels and mostly remained upstream (Figure 48). On H&E and endothelial cell (CD31) immunostaining images, P-LE-embolized FA showed significantly reduced vascular cellularity and CD31 expression, indicating the complete and rapid embolization of FA and vessel wall ablation using P-LE (Figures 40N and 40O). Next, FA embolization was repeated in heparinized (250U / kg) rats to evaluate P-LE embolization in an anticoagulated state. In heparinized rats with significantly increased ACT (Figure 49A), embolization was consistently successful indicating that coagulation is not necessary for P-LE embolization to occur (Figures 49B – 49G). These findings suggest that the P-LE formulation can reach the microvasculature when the embolic material is delivered in upstream larger blood vessels and successfully embolizes them. P-LE in porcine renal embolization model To determine whether P-LE can reach the distal vasculature and embolize them, a porcine kidney embolization model was used because of its characteristic hierarchical vascular network. Under real-time fluoroscopic guidance, a microcatheter was navigated to the proximal portion of the main lower pole renal artery and approximately 2 cc of the P-LE was injected over 30 seconds (Figure 41A). During injection of P-LE, Figure 41A demonstrates clear visibility of the embolic agent and opacification of the distal parenchymal vessels in the cortex suggesting that it is reaching the intended smaller vessels. Right after P-LE delivery, DSA was performed, showing the absence of flow to the lower pole of the kidney, indicating instant and complete embolization. At necropsy, a gross examination of the kidney revealed a geographic demarcation of the embolized lower pole of the kidney; this was more noticeable in the bisected kidney showing the dark discoloration of the embolized regions of the lower kidney segment (Figure 41B). P-LE embolized renal arteries were also distinctively visible on gross inspection (Figure 50). Detection of indocyanine green (ICG) fluorescence overlapped with the discolored portions of the kidney confirming that the ICG signal was delivered to the lower pole kidney via P-LE (Figure 41C). On histology, more than 50% of the embolized vessels measured up to 100 microns in diameter with 30% 50 microns or less suggesting that P-LE is an effective embolic agent of microvessels (Figure 41D). CD31 staining showed normal endothelium cellularity in the untreated controls, while vessels embolized with P-LE demonstrated absence of CD31 detection with significant vessel wall ablation (Figures 41E and 41F). As an indirect measure of successful embolization, immunostaining for hypoxia-inducible factor- 2α (HIF-2α) was performed suggesting detection of hypoxia surrounding embolized blood vessels (Figure 51). To further show that P-LE can reach distal microvessels and embolize them, P-LE was injected into the splenic artery; DSA images in Figure 52 clearly showed that P-LE successfully embolized the splenic sinusoids while maintaining flow in the main branch splenic artery. P-LE in anti-coagulated canine renal embolization model To simulate the clinical scenario of anticoagulation or coagulopathy, canines received IV heparin (250U / kg) resulting in a significantly increased ACT (Figure 53). Using a combination of a guide catheter and a microcatheter, P-LE was slowly injected from the main renal artery. Post-embolization angiography demonstrated successful embolization of the entire kidney using P-LE (Figure 42A). On IVIS imaging, ICG distribution was seen across the parenchyma of the renal cortex, suggesting that the P-LE likely reached the distal target microvessels (Figure 42B). Gross evaluation of the bisected kidneys clearly demonstrated embolized vessels (Figure 42C) and on histology, uniform embolization of sub-mm microvessels as small as 50 microns in diameter were noted, consistent with the porcine model of embolization (Figure 42D). Representative H&E images and immunostaining for CD31 showed that while untreated control vessels had normal cellularity and CD31 expression, the arteries that received P- LE showed absence of CD31 detection and reduced cellularity suggesting vessel wall ablation (Figures 42E – 42H). P-LE in porcine survival renal embolization model Porcine survival experiments were performed to evaluate the durability and superiority of P-LE embolization compared to the clinically used microsphere embolization (EMBOSPHERE®). Beads were suspended in a solution with equal amounts of IOH and ICG and then injected to stasis into the upper pole of the kidney; the lower pole of the same kidney received P-LE (Figure 43A). Subsequent DSA imaging demonstrated successful embolization. DSA imaging at day 14 revealed recanalization of the upper pole renal artery that received EMBOSPHERE®, consistent with the clinical experience. In contrast, the lower pole that received P-LE remained embolized at day 14 suggesting permanent embolization (Figure 43A). At necropsy, discoloration of the P-LE embolized tissue overlapped with ICG detection suggesting persistent delivery of the ICG to the renal cortex (Figures 43B and 54). On histology, the renal cortex that received EMBOSPHERE®showed mostly viable tissue with patent microvessels, while tissue that received P-LE demonstrated uniform embolization with >60% of arteries up to 100 microns containing intra-luminal P-LE and ablation of the parenchyma tissue (Figures 43C – 43E). On histology and immunostaining for CD31, vessels embolized with P-LE demonstrated significantly reduced vascular cellularity and absence of CD31 detection. No notable difference was observed with MPO immunostaining in the perivascular space; however, significant MPO staining was observed in the renal capsule, which is consistent with the renal capsular region having its own independent blood supply (Figures 43E, 43F, and 55). Trichome images of microvessels and their quantitative analyses showed significantly lower collagen content in P-LE-embolized vessels further confirming the permanent obstruction of blood flow (Figures 43E and 43F) (Coats et al., Circulation, 95:1293 (1997)). P-LE demonstrated no toxicity up to day 14 compared to baseline and immediately post-embolization. Oxygen saturation, ACT, CBCs, and serum biochemical analysis showed no unusual changes suggesting biocompatibility of P-LE (Figure 56). Evaluation of P-LE in acute porcine hemorrhage model To determine the efficacy of P-LE in embolizing acutely bleeding microvessels, porcine acute microvascular hemorrhage models were created in the kidney, liver, and stomach. Once the bleeding was confirmed by contrast extravasation, the microcatheter was selectively navigated to the bleeding site, and P-LE was injected to achieve instant embolization and treatment of the hemorrhage. Post-embolization fluoroscopic imaging demonstrated that the embolization was successful with rapid hemostasis (Figure 43G). These experiments indicate that in an acute scenario, P-LE can successfully treat hemorrhage. P-LE susceptibility tests to drug-resistant pathogens In clinical practice, abscess formation is an uncommon but potentially fatal long-term complication of embolization procedures (Bilbao et al., Semin INtervent Radiol, 23:126 (2006)). To determine whether P-LE can prevent potential post-embolization related infections, susceptibility tests were performed using some of the most antibiotic-resistant bacteria isolated from patients. P-LE demonstrated significant antimicrobial effects on all drug-resistant superbugs including, methicillin- resistant Staphylococcus aureus (MRSA; MIC, 0.097%), vancomycin-resistant Enterococcus faecium (VRE; MIC, 0.39%), and carbapenem-resistant Enterobacterales (CRE; MIC, 0.39%), Clostridioides difficile (C. difficile; MIC, 0.39%), and Candida auris (C. auris; MIC, <0.049%) (Figure 57). These exceptional antimicrobial effects of P-LE suggest that while conventional embolics may be at risk of procedural infections, P-LE may protect patients from potential septic complications. Example 3: Catheter-directed ionic liquid embolic agent for rapid portal vein embolization, segmentectomy, and bile duct ablation Materials and methods IL preparation Ionic liquids were synthesized in varying molar ratios of geranic acid (Sigma-Aldrich, St. Louis, MO) and choline bicarbonate (Sigma-Aldrich, St. Louis, MO). Briefly, HPLC grade acetone (Sigma Aldrich, St. Louis, MO) was added to a predetermined amount of geranic acid to achieve a final concentration of 30% (v / v) in a dry ice bath containing acetone reaching -80°C. The GA purification process was performed by repeated cycles of recrystallizing and removal of the acetone solvent and the associated impurities. The resulting purified geranic acid was then allowed to warm up to room temperature. Subsequently, an appropriate amount of choline bicarbonate was added to the purified geranic acid to achieve a specific molar-ratio (e.g., IL 3:1, where three moles of choline bicarbonate was added to one mole of GA). The reaction mixture was magnetically stirred overnight to complete the reaction, yielding IL compounds with carbon dioxide (CO2) and water (H2O) as side byproducts. Next, the material was transferred into a glass flask and evaporated at 50°C using a rotary evaporator (Rotavapor, Buchi, New Castle, DE) set at 150 rpm and 370 mbar vacuum to remove excess acetone and water molecules from the IL preparation. Diffusion assay To assess the diffusion potential of therapeutic agents after solubilizing in varying molar ratios of IL, two molecules were selected for testing due to their inherent fluorescent properties and net ionic charges; including doxorubicin (Pfizer, New York, NY) as a positively charged molecule, and indocyanine green (ICG, United States Pharmacopeia, North Bethesda, MD) as a negatively charged molecule.3-D printed cross inserts with centrally located cylinder-shaped attachments were positioned on top of each well of a 6-well plate to ensure equal loading capacity and homogeneous diffusion across the tested formulations. A tissue-mimicking matrix was created by dissolving agarose powder (2% w / v, Roche Diagnostics GmbH, Mannheim, Germany) in PBS (Thermo Fisher Scientific) pH 7.4 solution that was melted through boiling and subsequent cooling to 60°C. Twelve mL aliquots of the 2% melted agarose was poured into each well and left to solidify at room temperature, then the inserts were gently removed forming a centrally located well. Subsequently, 200 μL aliquots of IL formulations containing one of the three different molar ratios of IL (IL3:1, IL1:1, IL1:3) at set concentrations of 10%, 30%, 50%, or 70%, and equal amount of ICG (0.25 mg / mL), or doxorubicin (1 mg / mL) were added to designated wells. The area of diffusion was monitored using fluorescent imaging at 1-, 4-, 12-, and 24- hours using the IVIS Spectrum In Vivo Imaging System (PerkinElmer Inc., Waltham, MA) using appropriate sets of filters (740 / 850 nm emission / excitation filter was used to detect ICG and 460 / 560 nm emission / excitation filter was used to track doxorubicin). The diffusion area of fluorescence detection was quantified at a standardized threshold and scale values using the Living Image®software (PerkinElmer Inc.). Each measurement was replicated six times for each formulation. Cell culture The human cell lines, SNU478 (ampulla of Vater adenocarcinoma), HepG2 (hepatocellular carcinoma), and HUCCT1 (cholangiocarcinoma) were cultured in 75-mm2flasks (Corning Inc., Corning, NY) at 37°C and 5% CO2in a humidified chamber using Iscove's Modified Dulbecco's Medium (IMDM, Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (FBS, Gibco, Grand Island, NY), 1 mM sodium pyruvate (Gibco), 0.1 mM nonessential amino acids (NEAA, Gibco), 100 IU / mL penicillin (Thermo Fisher Scientific), and 10 µg / mL streptomycin (Thermo Fisher Scientific). The human umbilical cord vein endothelial cells (HUVEC) were cultured in Endothelial Cell Growth Medium (EGM-2, Lonza, Morriston, NJ). The SB1 mouse cholangiocarcinoma cell line was cultured in 75-mm2flasks using Dulbecco's Modified Eagle Medium (DMEM, Thermo Fisher Scientific) supplemented with 10% FBS, 100 IU penicillin and 10 µg / mL streptomycin as previously described (Albadawi et al., Sci Transl Med, 13 (2021)). All cell lines (SNU478, HUCCT1, HepG2, HUVEC) were obtained from the American Type Culture Collection (Manassas, VA). In vitro cytotoxicity assay SNU478, HUCCT1, HepG2, HUVEC, and SB1 cell lines were seeded into 96-well cell culture plates (CellTreat Scientific Products, Pepperell, MA) at a density of 5 x 103cell / well in growth medium and then incubated for 24 hours at 37°C and 5% CO2. After incubation, the medium was removed and replaced with 100 µL growth medium aliquot containing serially diluted ILs or LEAD, generating a concentration gradient ranging from 0.02% to 10%. Following a 24-hour incubation, the experimental medium was removed, and the wells were washed twice with 150 µL of PBS solution. Cell viability was assessed using the water-soluble tetrazolium-1 (WST-1) according to the manufacturer's instructions (Cayman Chemicals, Ann Arbor, MI). A 100 µL of growth media supplemented with 10 µL WST-1 reagent mix was added to each well then allowed to react for up to 2 hours inside the humidified cell culture incubator at 37°C and 5% CO2. The optical density in each well was acquired using a microplate reader (SpectraMax iD5, San Jose, CA) at a 450 nm wavelength. The relative viability rate was calculated as follows: Viability (%) = (1-ODtreated / ODcontrol) × 100%. Dose-response fractional viability plots were used to calculate IC50values for each cell line using the Prism Software ver.9 (GraphPad, San Diego, CA). The experiment was replicated eight times for each concentration. Live / dead cell assay To visualize the cytotoxic effect of ILs on different cells, 2x103cells per were suspended in 400 μL growth medium and seeded into a slide chamber (Labtek Inc., Vancouver, WA). After a 24-hour treatment period, the medium was removed, and 400 µL of media containing 0% IL3:1, 0.31% IL3:1, or 0.63% IL3:1 was added to the designated slide chambers. Following a subsequent 24-hour incubation, a cell viability reagent (Thermo Fisher Scientific, Waltham, MA) was added to each chamber and incubated for 15 minutes. Fluorescent microscopy images of viable cells (stained blue) and nonviable cells (stained green) were captured using an inverted fluorescent microscope with the appropriate filter sets (EVOS FL Auto 2, Thermo Fisher Scientific, Waltham, MA). Drug synergy assay A synergy assay was performed on SNU478 cells to investigate the potential interaction between low concentrations of IL3:1 and cisplatin (Calbiochem, Darmstadt, Germany), one of the most commonly used chemotherapeutics for patients with cholangiocarcinoma. Initially, the IC50value of cisplatin for the SNU478 cell line was determined using the aforementioned in vitro fractional cell viability method. Subsequently, the concentrations spanning the IC50values were selected to create treatment formulations. These formulations included individual drugs (IL or cisplatin) and pairwise treatments with all possible combinations of IL3:1 and cisplatin, using the same concentrations as in the single treatments. This process resulted in a 5x5 matrix for each tested drug, with pairwise concentrations interpolated from the fitted Hill curves of each single treatment viability plot using the Loewe model. Loewe synergy plots were generated using the Combenefit software, and synergy values were calculated using the HSA synergy and antagonism assay. ATR-FTIR characterization Aliquots of synthesized IL3:1, GA, and choline bicarbonate were analyzed using attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy (Lumos II equipped with an Alpha II extension, Bruker, Kontich, Belgium) to confirm chemical composition and detect potential changes in chemical bonds. Infrared vibrational spectra in absorbance mode covering the spectral range of 4000 to 500 cm-1wavenumbers with a resolution of 4 cm-1, were recorded for each material. Each test was repeated at least three times, and the results were graphically illustrated using the OPUS software (Bruker, Billerica, MA). Zeta potential and conductivity measurements The zeta potential and conductivity of IL3:1 at varying concentrations ranging from 6.3% to 70% loaded into disposable folded capillary cells was measured using the Zetasizer ultra instrument (Malvern Panalytical Ltd., Malvern, United Kingdom). The acquired data were analyzed using ZS Xplorer software (Malvern Panalytical Ltd.). Each measurement was replicated four times for each concentration. Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) To assess the stability of Nivolumab (Bristol Myers Squibb, Princeton, NJ) after mixing with ionic liquid-based formulations, aliquots containing 1 mg / mL nivolumab in PBS or IL3:1 were incubated at 37°C for up to 28 days. At predetermined time point (0, 1, 7, 14, or 28 days), one aliquot was removed and mixed with protein denaturing solution containing 2-mercaptoethanol (Sigma-Aldrich) and Laemmli protein sample buffer (1610747, Bio-Rad) and boiled for five minutes. Subsequently, samples containing 5 µg of Nivolumab were loaded into the acrylamide gel (Bio-Rad, Hercules, CA) and the proteins were fractionated using electrophoreses at 100 Volts. Next, proteins in the gel were transferred onto a 0.2 µm polyvinylidene fluoride membrane (Bio-Rad) using the Trans-Blot Turbo system (Bio-Rad). The protein bands were visualized using the Gel Dox XR+ imaging system (Bio-Rad) and the bands intensities were quantified using the ImageJ software (National Institutes of Health, Bethesda, MD). Assessing radiodensity For visualization during percutaneous endovascular interventions, IOH (GE HealthCare Systems, Chicago, IL) contrast agent was incorporated into the LEAD formulation. LEAD containing increasing IOH concentrations ranging from 0% to 100% were prepared by mixing IOH with IL3:1. The mixture was loaded into 1-mL Medallion syringes (Merit Medical Systems Inc., South Jordan, UT), then imaged high-level fluoroscopy acquired using an OEC 3D Fluoroscope (GE HealthCare). Pixel intensity in each syringe was calculated using ImageJ software (National Institutes of Health, Bethesda, MD). ICG stability assessment To investigate the stability of ICG in the LEAD formulation, a comparative analysis of ICG stability was conducted at 37°C; ICG solubilized in water (Control) and ICG solubilized in LEAD. The fluorescence intensity measurements were taken at days 0, 1, 2, 3, 5, and 7 using a fluorescence spectrophotometer. Injection force measurements The injection force was measured using a mechanical tester (Instron 5942, Instron, Norwood, MA) by applying compression forces to 1-mL Medallion syringes filled with various samples being injected at a flow rate of 1 mL per minute through a 150 cm 2.8F catheter (Boston Scientific, Marlborough, MA) into a PBS solution to mimic the delivery into the bloodstream. Flow curves were plotted using the Bluehill version 3 Software (Instron). Break-loose and injection force values were measured in three sets for every formulation tested. Rheometry studies The rheological measurements were performed using a rheometer (MCR 302, Anton Paar, Torrance, CA) to evaluate the viscosity of the IL3:1 or LEAD. The assessment was performed at a shear rate ranging from 100to 103s-1at 37°C. The gap between the sandblasted 25 mm aluminum shaft plate and the lower aluminum plate was kept at 1 mm during throughout the tests. Each measurement was replicated three times. Transwell diffusion assay The diffusivity and cytotoxicity of the ionic liquid were assessed across a small-size pore tissue- like barrier that mimics the in vivo conditions. To perform the experiment, 4x105SNU478 cells were seeded in the lower chamber of a 12-well transwell plate. An upper chamber insert fitted with a 0.4 μm pore size polycarbonate membrane (Corning Inc.) was mounted on top of each well. To replicate the pore size of the perivascular matrix and cellular layers in the portal vein, the membrane in the upper chamber was initially covered with a 500 μL aliquot of 2% melted agarose and allowed to solidify at room temperature. Subsequently, a 200 μL pig aliquot blood was added on top of the agarose hydrogel in the upper chamber to simulate the intravascular environment. A 200 μL of LEAD or saline + ICG aliquots were then added on top of the blood in designated upper chambers to mimic the endovascular delivery. As a control, the upper chamber was treated with a formulation containing all LEADS components except the ionic liquid. After a 24-hour incubation at 37°C and 5% CO2, cell viability and ICG fluorescence intensity were evaluated in the lower chamber, as described elsewhere (Adlbadawi et al., Sci Transl Med, 13 (2021)). Cell migration assay To investigate the impact of LEAD on cellular migration using sublethal concentrations of LEAD that induce sub-IC50cytotoxicity. SNU478 were suspended in growth medium and 5x104cells were seeded inside the 2-well silicon insert within a 35-mm µdish (Ibidi, Grafelfing, Germany) to create a defined 500 µm cell-free gap. After a 24-hour incubation, the inserts were removed, and a medium containing varying concentrations of LEAD (0%, 0.31%, 0.63%) was added to the µdish. Serial phases contrast digital images were acquired using an EVOS FL Auto-2 microscope. The experiment was terminated when the gap in the control µdish treated with 0% LEAD was closed. The area of migration was calculated using the QuPath open-source software. Cell adhesion assay To investigate the impact of LEAD on cell adhesion, 1x106SNU478 cells were seeded into the wells of a 6-well plate in a 2 mL growth medium containing various LEAD concentrations. The control wells had growth medium alone. The cells were then observed hourly under a microscope until near-full adhesion in the control group was observed. The wells were gently washed twice with PBS solution to remove unattached cells, and 1 mL of fresh medium was added to each well. The cells were then imaged at 20x magnification using an EVOS FL Auto 2 microscope and the number of adherent cells in standardized 8 fields acquired in each well was counted using the QuPath software. Sterility test To confirm the sterility of LEAD preparation prior to in vivo testing, of LEAD solution was aseptically diluted in a fresh Luria-Bertani Broth (LB Broth, Fisher Scientific, New Lawn, NJ) at 1:14 v / v ratio in a sterile tube. The mixture was then incubated at 37°C inside a shaker incubator set a 180 RPM for 24 hours. Positive controls consisted of LB Broth inoculated with 107CFU mL-1of chemically competent E. coli bacteria, while negative controls included LB Broth without the addition of the LEAD sample. After 24 hours, 100 µL aliquots from each tube was transferred into a 96-well plate, and the optical density was measured at 600 nm (OD600) using a microplate reader to assess turbidity, providing a quantitative measure of bacterial growth. To further validate the sterility of LEAD preparation, 100 μL LEAD or E. coli samples were inoculated onto agar plates (Fisher Scientific, New Lawn, NJ) and incubated at 37°C for 24 hours. Subsequently, the agar plates were observed and photographed at 24 hours. The sterility experiments were conducted in quadruplicates. Antimicrobial Assay The antibacterial potential of LEAD was evaluated by measuring the OD600values of E. coli bacteria culture (25922, ATCC) using a plate reader. The E. coli medium was appropriately diluted in fresh LB broth to achieve an OD600value of 0.7 representing 5.6x108bacterial cells mL-1. Subsequently, the E. coli samples were mixed with predetermined ratios of LEAD and incubated for 24 hours at 37°C in an orbital shaker incubator set at 180 rpm. After 24 hours, 100 µL aliquots of each sample were transferred to a 96-well plate, and the absorbance values at 600 nm were measured by using a microplate reader (SpectraMax iD5, Molecular Devices). To illustrate the antibacterial effect of LEAD on the number of E. coli colonies, the samples were diluted x104folds. A 100 µL aliquot of the diluted samples including the controls were inoculated onto agar plates and incubated at 37°C for 24 hours. After 24 hours, the plates were inspected for bacterial growth and photographed. Paralleled experiments were performed to assess the antibacterial potential of LEAD against highly resistant bacteria, including the methicillin-resistant S. aureus (MRSA), vancomycin-resistant Enterococcus (VRE), carbapenem- resistant Enterobacteriaceae New Delhi metalloprotease-1 (CRE-NDM-1), Klebsiella pneumoniae (K. pneumo), extended-spectrum beta-lactamase E. coli (ESBL E. coli), Pseudomonas aeruginosa, and Enterobacter cloacae. Briefly, each bacterial culture was diluted in sterile normal saline to achieve an OD600value of 0.6 representing 4.8x108bacterial cells mL-1. An equal volume of bacterial culture was mixed with LEAD at various concentrations in sterile tubes and incubated at 37°C for 24 hours. A 100 µL aliquot from each culture was inoculated onto designated six locations in the blood agar plates (Thermo Fisher Scientific). The plates were then incubated at 37°C for 24 hours and then photographed to assess the absence or presence of bacterial growth. Assessment of LEAD ablation and drug delivery potential in ex vivo human tissues Freshly harvested human tissues including, non-fibrotic liver (n=4 for LEAD, n=4 for control), fibrotic liver (n=4 for LEAD, n=4 for control), renal cell carcinoma (n=3 for LEAD, n=3 for control), and lung adenocarcinoma (n=1 for LEAD, n=1 for control) were obtained for ex vivo testing. A 50 µL aliquot of LEAD loaded in a syringe was injected into the tissue core through a 21-gauge needle and then incubated for up to 24 hours inside a humidified chamber at 37°C. At predetermined time intervals, the injected tissues were imaged to assess drug diffusion based on ICG fluorescence included in LEAD using the IVIS imaging system. Subsequently, the tissues were fixed in 10% buffered formalin (Fisher Scientific, Waltham, MA) and processed for histological examination.4 µm serial histology sections were stained hematoxylin & eosin (Thermo Fisher Scientific) to assess morphology or immunostained for Nivolumab detection. Assessing the effect of LEAD on blood hemolysis and cell counts To verify the effect of various concentrations of LEAD on blood hemolysis, citrated whole pig blood (Innovative Research, Inc.) was centrifuged at 2000 rpm for 10 minutes to remove plasma. The sedimented blood cells were then resuspended to the original volume using isotonic sodium chloride solution and then diluted into aliquots with PBS using a 1:19 v / v ratio. Different diluted aliquots were incubated with various concentrations of LEAD, PBS (negative control), or ACK cell lysing buffer (positive control, KD Medical, Columbia, MD). After a 1-hour incubation period at 37°C and 5% CO2, the samples were centrifuged at 1500xg for 10 minutes at room temperature. A 100 µL supernatant aliquot was transferred into a 96-well plate, and the absorbance values were measured at 450 nm wavelength using a microplate reader. In parallel, blood smears on positively charged glass slides were prepared and stained with Wright staining (Camco Quik Stain) according to the manufacturer’s protocol and then observed under a brightfield microscope to visualize blood cells. Additionally, to investigate the effect of LEAD on pig blood count, whole pig blood aliquots were combined with LEAD at various concentrations ranging from 0.1% to 10% and then incubated for one hour inside a humidified chamber at 37°C and 5% CO2. A complete blood count was performed using a hematology analyzer (HemaTrue, Heska). Assessing the effect of LEAD on thrombosis To investigate the effect of LEAD on blood coagulation, prewarmed uncoagulated citrated pig blood was mixed with varying amounts of LEAD to achieve final concentrations of 0.1%, 1%, 2%, 5%, and 10%. Next, blood coagulation was initiated by adding a 1:30 v / v ratio of 0.2 M CaCl2solution. After brief mixing, 1 mL aliquots of each activated blood sample were treated with an appropriate amount of LEAD and then quickly transferred to multiple wells in a 12-well plate. In parallel, control samples consisted of activated whole pig blood aliquots without treatment. Clot formation was monitored time by removing excess liquid at 1, 3, 5, 7, 10, 20, or 30 minutes and recorded when liquid can no longer be removed. Additionally, ACT was performed on calcium-induced coagulation of citrated whole pig blood after mixing with LEAD at predetermined concentrations of 0.1%, 1%, 2%, 5%, 10%. The ACT of the mixtures was recorded using i-STAT blood analyzer (Abbott Laboratories, Chicago, IL) and a Celite ACT cartridge (Abbott Laboratories, Chicago, IL). Hemorheology measurements To illustrate changes in blood clotting behavior induced by LEAD, time-dependent change in clot modulus if activated pig blood treated with LEAD was monitored using a rheometer (Anton Paar MCR 302). Citrated whole pig blood samples were mixed with LEAD at predetermined concentrations of 0.1%, 1%, 2%, 5%, and 10% followed by the addition of 0.2 M CaCl2to the LEAD + blood mixtures at 1:30 v / v ratio. A 0.7 mL aliquot of the mixture was added to the lower plate of the rheometer. The gap between the 25 mm sandblasted aluminum shaft plate and the lower plate was kept at 1 mm. A solvent trap filled with water was used to provide a humidified environment. Flow curves and amplitude sweeps at 10 rad s−1were generated at 37°C for 30 minutes. The storage modulus (G’) values were recorded for each test at the end of the 30-minute interval with each measurement repeated three times. In vivo portal vein ablation and drug delivery in a rat model Two groups of Sprague Dawley rats were anesthetized using continuous inhalation of 2% isoflurane in 100% Oxygen using a nose cone. The rats were positioned supine on a warming platform with temperature regulation to 37°C using a rectal probe. A midline laparotomy exposed the liver, and wet cotton-tipped applicators were used to elevate the right and left medial liver lobes, exposing the portal vein alongside the hepatic artery and common bile duct. Access to the portal vein was established using a 25-gauge needle, followed by the insertion of a 1F catheter (World Precision Instruments, Sarasota, FL), carefully advanced inside the vein. The treatment group received a 100 µL endovascular injection of LEAD containing 70% IL3:1, 0.25 mg / mL ICG, 20% IOH, and 1 mg / mL nivolumab through the catheter. In contrast, the control group received a 100 µL endovascular injection of a solution containing 70% PBS, 0.25 mg / mL ICG, 20% IOH, and 1 mg / mL nivolumab. After the injection, the catheter was removed, and hemostasis was achieved by applying mild pressure to the portal vein with a cotton-tipped applicator. The subcutaneous and dermal layers were approximated using 5-0 Vicryl sutures, and the rats were kept under inhalation anesthesia with continuous monitoring for one hour, followed by euthanasia. Liver tissue was then explanted, and near-infrared fluorescence imaging was performed using the IVIS Spectrum In Vivo Imaging System at a 740 / 850 nm wavelength to assess ICG distribution on intact liver and after bisection of the liver lobes. After fluorescence imaging, the liver tissue was fixed in 10% buffered formalin and processed for histopathology. Renal artery embolization in a non-survival porcine model Percutaneous renal artery embolization procedure was performed on Yorkshire / Landrace pigs weighing 50-55 kg. Pre-procedure anesthesia was induced by 5 mg / kg of tiletamine-zolazepam (TELAZOL®, Zoetis, Parsippany-Troy Hills, NJ), 0.02 mg / kg glycopyrrolate, and 2 mg / mL xylazine. After endotracheal intubation, the pigs were placed supine on an x-ray compatible table and anesthesia was maintained with the 1-2% isoflurane inhalation throughout the procedure. The percutaneous access to the iliac artery was achieved under ultrasound imaging guidance (Butterfly iQ+, Butterfly Network Inc., Guilford, CT). A 0.018-inch access wire (Cook Medical) was initially placed, which was later replaced with a 0.035-inch wire to introduce a 5F catheter sheath. Over this sheath, a 5F Torcon angiographic catheter (Cook Medical) was advanced to the abdominal aorta. The catheter tip was positioned in one of the main renal arteries, confirmed by real-time digital subtraction angiography (DSA). A 3 mL aliquot of radiopaque LEAD was then delivered to the renal artery under real-time fluoroscopy. Following the delivery of LEAD, DSA was repeated to confirm the embolization of the renal artery vascular network. At 1-hour post-embolization, the pigs were euthanized, and the renal tissues were harvested. The tissues were bisected across their mid-sagittal plane for fluorescence imaging using the IVIS Spectrum In Vivo Imaging System at a wavelength of 740 / 850 nm to visualize ICG distribution. After IVIS imaging, the tissues were fixed in 10% buffered formalin and processed for histopathology. In vivo portal vein embolization, ablation and drug delivery in a porcine model The portal vein embolization procedure was performed on Yorkshire / Landrace pigs weighing 50- 55 kg. Anesthesia was induced by the intramuscular injection of 5 mg / kg of tiletamine-zolazepam, 0.02 mg / kg glycopyrrolate, and 2 mg / mL xylazine. After endotracheal intubation, the pigs were positioned supine on an x-ray-compatible-table and anesthesia was maintained with 1-2% isoflurane inhalation throughout the entire procedure. Following sterilization of the surgical field, the portal vein was visualized using ultrasonography. Upon percutaneous access to the portal vein, a 0.018-inch access wire (Cook Medical) was placed in the vein. Following the replacement of the 0.018-inch access wire with a 0.035-inch Glidewire (Terumo), a 5F catheter (Yueh, Cook Medical, Bloomington, IN) was advanced over the wire. The catheter was maneuvered to reach the portal vein branch supplying the left lateral lobe of the porcine liver under fluoroscopic guidance. IOH contrast was injected to confirm the catheter location in the portal tract under fluoroscopy. Subsequently, 10 mL of LEAD was delivered to the portal vein branch supplying the left lateral lobe of the porcine liver for embolization, ablation, and drug delivery purposes. Embolization was achieved in less than a minute and was confirmed by repeat fluoroscopy after LEAD delivery. Blood samples and ECG readings were taken before and after embolization.100 U / kg of heparin was administered intravenously to prevent coagulation. In the non- survival studies, pigs were euthanized within 1 hour after portal vein embolization. In a separate cohort, pigs survived for 7 days followed by angiography of the portal vein before euthanasia. At necropsy, the liver tissues were harvested for gross imaging and near-infrared fluorescence imaging using the In Vivo Imaging System at a wavelength of 740 / 850 nm for ICG visualization on the intact liver tissue and after trans-sectioning of the embolized and the untreated control lobes. After imaging, the liver tissue was fixed in 10% buffered formalin and processed for histology. Mouse model of cholangiocarcinoma The mouse SB1 cholangiocarcinoma cell line was used to induce ectopic tumors in C57BL6 male mice weighing 25-30 grams. To create the model, SB1 cells were cultured in DMEM growth medium containing 10% FBS (v / v), and penicillin streptomycin antibiotics in 75-mm2flasks. Before tumor inoculation, confluent SB1 cells underwent PBS washing, enzymatically dissociated using 0.25% Trypsin-EDTA followed by washing in growth medium. The cells were then diluted in supplement-free DMEM to create a one million cells mL-1suspension. Subsequently, a 50 μL aliquot of SB1 cell suspension was subcutaneously injected into the lower right flank of each animal. A cotton-tip applicator was used to apply gentle pressure to prevent backflow. After the tumor inoculation, the mice were monitored twice a week to detect tumor formation. Once tumors are detected, tumor growth was evaluated using a caliper to measure maximum diameters in two planes: superior-inferior (SI) and lateral- medial (LM). Tumor volumes (TV) were calculated using the formula: TV=(4πx(SI)x(LM)x((SI+LM)÷2)÷3. Additionally, an ACUSON S2000 ultrasound system with a multifrequency linear transducer (9L4, 9.0 MHz) was employed to visualize tumor borders in a three- dimensional plane, and relevant 2D images were captured in B-Mode. Upon reaching a tumor volume of approximately 0.1 cm3, mice were randomly assigned to treatment or control groups. When the tumor volume reached 0.5 cm3, control group mice were sacrificed, while the treatment group received an intratumoral injection of LEAD using a 25-gauge needle under ultrasonography guidance. Mice in the treatment group were euthanized an hour after injection. During necropsy, tumor tissues were explanted, fixed in 10% buffered formalin, and processed for histology. Histological evaluation The harvested tissues were fixed in 10% buffered formalin and embedded. Paraffin blocks were sectioned at 4 μm and mounted on positively charged glass slides (Fisher Scientific, #12-550-15, Pittsburgh, PA). The slides were baked at 60°C for 30 minutes, deparaffinized (2 x 100% xylene, 3 x 100% ethanol, 1 x 95% ethanol, 1 x 80% ethanol, 1 x 70% ethanol, 5 minutes each), and rehydrated in double-distilled water for 5 minutes. Serial sections were stained with hematoxylin and eosin (H&E, 7111 and 7221, ThermoFisher Scientific) to visualize tissue morphology. To visualize cholangiocytes in liver tissues, rat and pig liver sections were incubated with a rabbit anti-Rat CK7 IgG (AB181598, Abcam, 1:1000). To detect endothelial cells, rat and pig liver sections were incubated with a rabbit anti- mouse CD31 (ab182981, 1:1000, Abcam). To show hypoxia in kidney tissues, pig kidney sections were incubated with a rabbit anti-mouse HIF-2α (ab109616, 1:1000, Abcam). To observe infiltrating monocytes and macrophages, tissue sections were incubated with polyclonal rabbit anti-CD68 IgG (ab125212, Abcam, 1:250) (Albadawi et al., Sci Transl Med, 13 (2021)). To detect infiltrating granulocytes, tissue sections were incubated with anti-myeloperoxidase antibody (ab208670, 1:500, Abcam) and goat anti-rabbit IgG (HRP) (ab97051, Abcam, 1:500) was used as a secondary antibody. The checkpoint inhibitor, anti-PD-1 IgG4 (nivolumab) was visualized by incubating tissue sections with 5 μg / mL of the biotin-conjugated monoclonal rabbit anti-human IgG4 (ab238617, Abcam, Cambridge, MA). An EVOS FL Auto-2 microscope was used to obtain digitally stitched or individual micrographs at 20x magnification. Morphometric analysis was performed to count the number of CK7+, CD31+positive cells at each section in the rat and pig liver. These were conducted in twelve randomized fields per specimen. Additionally, the connective tissue of the liver capsule in the survival porcine studies was visualized using Picrosirius red (ab150681, Abcam) stained sections. All measurements and cell counts were performed using Qu Path software. Blood collection and analysis in pigs Whole porcine blood and serum samples were collected immediately before embolization with LEAD and after 7 days survival period. The collected blood samples were tested for complete blood counts and activated coagulation time (ACT, Abbott Laboratories, Chicago, IL). Serum markers of organ function were analyzed using DRI-CHEM 4000 (Heska), and a presurgical panel cartridge (Heska, Loveland, CO), including creatinine (CRE), blood urea nitrogen (BUN), alkaline phosphatase (ALP), alanine aminotransferase (ALT), and glucose (Glu). Statistical analyses The data, unless otherwise specified, are presented as mean ± s.e.m. and are derived from a minimum of three replicate experiments. Statistical analyses were conducted using either a two-tailed unpaired Student’s t-test for comparisons between two groups or a one-way ANOVA with Tukey’s or Dunnett’s post-hoc tests for comparisons involving three or more groups, as deemed appropriate. A probability value (P) of less than 0.05 was considered statistically significant. All statistical analyses were performed using GraphPad Prism 9. Results Preparation and characterization of ILs and optimization of LEAD formulation To develop a potent embolic agent with both ablation and drug delivery properties, three IL formulations were synthesized with varying molar ratios between choline bicarbonate and geranic acid (3:1, 1:1 and 1:3) (Figures 58A and 58B). To identify the optimal IL candidate for the LEAD formulation, diffusibility tests were conducted using a 2% w / v agar tissue-like matrix with an average pore size of 140 nm (Pluen et al., Biophys J, 77:542 (1999)). This setup closely mimics the average pore size of human hepatic sinusoidal capillaries, which could be as large as 180 nm (Sari, J Angiogenes Res, 2:14 (2010)). To help assess the diffusion capacity of different IL candidates, fluorescent markers such as doxorubicin (Dox) and ICG were included in the mixture to allow detection using IVIS imaging. Diffusion studies showed that IL, with a 3:1 molar ratio, had the highest diffusion rate compared to 1:1 and 1:3 ratios at the 24-hour time point (Figures 58C, 65A, and 65B). The higher rate of ICG and doxorubicin diffusion in IL3:1 formulation can be attributed to the significant hydrophilic quaternary ammonium structure of choline, which may enhance the diffusion of surrogate drugs within hydrophilic matrices, such as agarose gel, and possibly within tissue extracellular matrix (Rantz et al., J Cell Sci, 123:4195 (2010); Jiang et al., Mar Drugs, 21 (2023); Sugiyama et al., Anal Chem, 87:11176 (2015)). Based on these results, IL3:1 was selected for further testing as diffusion capacity is an important feature for effective transvascular drug delivery and tissue ablation. Subsequently, diffusion analysis was repeated using various concentrations of IL3:1 (10, 30, 50, and 70 % w / v). The results indicated that enhanced diffusion is associated with an increased concentration of IL, suggesting IL's role in delivering payloads in a tissue-like matrix (Figures 66A and 66B). To evaluate the potential anti-cancer properties of IL, cell viability assays were performed using a range of cell lines, including those from human cholangiocarcinoma and liver cancer. Fractional viability studies showed the potency of IL against cancer cells with the 3:1 formulation demonstrating IC50values of 0.43% for SNU478, 0.44% for HUCCT1, 0.89% for HepG2, and 0.14% for SB1 cancer cell lines (Figures 58D and 67). The effect of IL3:1 was also tested on HUVEC, a human endothelial cell line, since LEAD was delivered into the portal vein, yielding an IC50value of 0.47% (Figure 67H). A live / dead cell assay was subsequently performed with IL concentrations near the IC50,confirming the viability assays (Figure 58E). The combined effect of IL mixed with cisplatin, a commonly used chemotherapeutic in cholangiocarcinoma treatment, was then investigated to see if synergy can enhance their cytotoxic effects at lower concentrations; cisplatin is known to have severe side effects such as hearing loss, kidney injury, and neuropathy (Oh et al., Lancet Gastroenterol Hepatol, 7:522 (2022)). The synergy between IL and cisplatin was seen at lower concentrations of both components (Figures 68A – 68D), suggesting a potential strategy for more effective and safer chemotherapy regimens in cholangiocarcinoma treatment. To further characterize IL3:1, Fourier transform infrared spectroscopy (FTIR) was used. This technique enabled the precise identification of chemical bond locations and their relative intensities (Figure 69). Subsequently, the impact of varying concentrations of IL3:1 on its zeta potential and conductivity was investigated. Concentrations ranging from 6.3% to 70% were tested, and the results showed a consistent increase in zeta potential from -39 mV to -5 mV (Figure 70A) and conductivity from 14.56 mS / cm to 41.31 mS / cm (Figure 70B). This trend suggests a progressive shift towards a more positive surface charge at higher IL3:1 concentrations. Nivolumab, the most used immunotherapy drug worldwide, was also included as a representative large protein-based drug. This integration was important for assessing the delivery capabilities of LEAD. To ensure the stability of nivolumab within this delivery framework, extensive stability testing was undertaken. This involved a 28-day incubation period of nivolumab in the embolic material (IL) at a constant temperature of 37ºC. Protein analyses showed no noticeable degradation of both heavy and light chain bands of nivolumab up to 28 days. (Figure 58F). For safe and effective administration and monitoring of LEAD during the procedure, IOH (Widmark, Proc (Bayl Univ Med Cent), 20:408 (2007)), an FDA-approved contrast agent, was included in the IL formulation to allow visibility of the injected material during fluoroscopy. Advanced fluoroscopic imaging demonstrated that IL formulations containing 20% IOH provided sufficient visibility (Figure 58G). This concentration was carefully optimized to balance both adequate visualization under fluoroscopy and maximal IL concentration within the LEAD formulation for tissue ablation. For tracking LEAD in tissue and to simulate the delivery of a molecular drug, the fluorescent dye, Indocyanine Green, was integrated into the embolic formulation. Next, the stability of ICG was assessed at 37°C, demonstrating that LEAD preserved ICG fluorescence up to 7 days, indicating LEAD’s role in stabilizing ICG vulnerable to rapid degradation (Figure 71). LEAD formulation, finalized as 70% IL3:1 (w / v) + 20% IOH (v / v) + 1 mg / mL nivolumab + 0.25 mg / mL ICG, showed similar IC50and diffusibility values between LEAD and IL3:1 alone (Figures 58H and 58I). The injection force of LEAD through a 2.8F microcatheter was measured to be 3.4 N, allowing easy hand-injectability (Figure 58J). The viscosity of LEAD at 37°C was found to be 20 mPa (Figure 58K). In vitro evaluation and anti-bacterial activity of LEAD For LEAD to reach tumor tissue when delivered into the portal vein, it must first interact with blood and then diffuse across the vessel wall and through the extracellular matrix to reach the tumor. To test this, a Transwell culture plate with a 400 nm pore size insert was used to simulate the hyperpermeable vasculature of solid tumors (Stylianopoulos and Jain, Proc Natl Acad Sci USA, 110:18632 (2013)). Following overnight incubation of SNU478 cholangiocarcinoma cells in the lower chamber, the surface of the upper chamber was first coated with 500 µL of agarose gel. Then, 200 µL of blood followed by 200 µL of LEAD or saline + ICG was added in the upper chamber. After 24 hours, only 0.01% of cholangiocarcinoma cells in the lower chamber were viable in the LEAD group compared to the control samples (Figure 59A). Furthermore, ICG fluorescence at the lower chamber in the LEAD group was significantly higher than in the control group, suggesting that IL facilitated delivery of ICG into the lower chamber (Figure 59A). These data demonstrate that LEAD was able to traverse multiple barriers such as blood, tissue-mimicking agarose gel, and the pores of the Transwell and maintain its cytotoxic potency in the lower chamber. Next, the effect of LEAD on migration and adhesion was investigated on cholangiocarcinoma cells. A migration assay was conducted using sub-IC50and IC50concentrations of LEAD. It was found that the migration of cancer cells was significantly reduced by 32.3% at a 0.31% LEAD concentration and by 74.9% at a 0.63% LEAD concentration (Figure 59B) at 24 hours. Furthermore, an adhesion assay was performed to assess the impact of LEAD on the adhesive potential of cancer cells at both sub-IC50and IC50concentrations. The results indicated a substantial decrease in cell adherence to the tissue culture plate, with reductions observed at 62.5% for 0.31% LEAD and 66.6% for 0.63% LEAD concentrations at 4 hours, respectively (Figure 59C). These findings, showing diminished migration and adhesion of cancer cells, highlight the anti-cancer properties of LEAD even at sub-lethal concentrations, suggesting that LEAD could still be effective at tumor margins. Embolization procedures can render tissues vulnerable to infection due to diminished blood supply. This is particularly evident in procedures like portal vein embolization, where complications such as bacterial cholangitis, hepatic abscess, or sepsis can occur (Yeom and Shin, Korean J Radiol, 16:1070 (2015)). Therefore, the antibacterial properties of an embolic agent are valuable in mitigating post- embolization infections. In this context, the antibacterial efficacy of LEAD was rigorously tested against E. coli and various highly drug-resistant microbes. The sterility of LEAD was first established through a controlled experiment; the OD600of LEAD was measured after a 24-hour incubation at 37°C and was found to be comparable to that of the negative control (LB broth), indicating no bacterial growth (Figure 59D). The antimicrobial effect of LEAD against E. coli, the most commonly isolated organism in cholangitis cases (Fomi et al., J Hepatobiliary Pancreat Sci, 25:3 (2018); Hu et al., Scientific Reports, 9:11947 (2019)), was then evaluated. The results showed that LEAD demonstrated a substantial bactericidal effect with an IC50value of 1.56% against E. coli (Figure 59E). Furthermore, LEAD achieved a dose-dependent decrease in the number of E. coli colonies on agar plates, showing a potent growth inhibition. Next, the scope of the antimicrobial assay was expanded to include antibiotic-resistant pathogens isolated from patients such as methicillin-resistant S. aureus (MRSA), vancomycin-resistant Enterococcus (VRE), Pseudomonas aeruginosa, carbapenem-resistant Enterobacteriaceae New Delhi metalloprotease-1 (CRE-NDM-1), Klebsiella pneumoniae (K. pneumoniae), Enterobacter cloacae, and extended-spectrum beta-lactamase E. coli (ESBL E. coli). Against these resistant pathogens, a pronounced antimicrobial effect was consistently shown by LEAD, with MICs ranging from 0.1% to 1.6% (Figures 59F, 59G, and 72). These findings suggest that LEAD could offer a dual benefit in embolization procedures: not only does it serve as an effective embolic agent, but it also has the potential to significantly reduce, or even eliminate, infection-related complications that are a risk with conventional embolic agents. This dual functionality of LEAD, especially its efficacy at diluted concentrations, marks an advancement in the field of embolization therapy, potentially increasing the safety and effectiveness of these procedures. Ablation and drug delivery properties of LEAD in human tissues To demonstrate LEAD’s ablation and drug diffusion potential in human tissues prior to in vivo studies, freshly resected non-fibrotic and fibrotic human liver tissues were directly injected with LEAD (Figures 60A and 60F). IVIS images of the tissues were taken at predetermined time points. Following a 24-hour incubation, the tissues were sectioned for gross and IVIS imaging. These demonstrated LEAD’s effective diffusion within liver tissues over time (Figures 60B and 60G), with histological analysis revealing significant ablation of liver tissue, characterized by an almost complete absence of cellularity (Figures 60C, 60D, 60H, and 60I). This points to LEAD’s capacity for extensive tissue penetration and effectiveness in tissue ablation. Furthermore, the successful delivery of nivolumab, a large protein-based immunotherapeutic, throughout the liver tissues was also shown by immunohistochemistry studies (Figures 60C, 60E, 60H, and 60J). The ability of LEAD to efficiently transport and disperse a therapeutic agent within the liver matrix is noteworthy, particularly in the context of fibrotic liver tissues. Fibrotic tissues are known for their dense fibrous composition and an abundance of extracellular matrix, which typically pose challenges for effective drug delivery (Kisseleva and Brenner, Nat Rev Gastroenterol Hepatol, 18:151 (2021)). These results affirm the potential of LEAD as a versatile embolic agent and underscore its capability to serve as a vehicle for delivering large protein-based therapeutics. This dual function is especially useful in the context of treating liver conditions, where both tissue ablation and targeted therapeutic delivery are often necessary. The ability of LEAD to perform these functions effectively, even in challenging fibrotic liver tissues, represents a significant advancement in the field of liver therapeutics and embolization technology. To test LEAD’s ablation and drug diffusion potential in cancerous human tissues, three freshly resected human renal cell carcinoma and one lung carcinoma tumor tissue were also injected with LEAD (Figures 73 and 74). IVIS images of the cancerous tissues were taken at predetermined time points, revealing LEAD’s ability to diffuse uniformly through the tumor tissues (Figures 73A, 73C, and 74A). Histological studies showed effective ablation and the concurrent delivery of nivolumab across the tumor tissues. (Figures 73B -73E and 74B – 74D). These results indicate that LEAD can effectively ablate and deliver large protein-based immunotherapeutics across tumor matrices. Portal vein embolization with LEAD in a non-survival rat liver model To evaluate LEAD’s capability to embolize, ablate, and deliver drugs, portal vein embolization was performed in a non-survival rat liver model (Figures 61A, 61B, and 75). Following a laparotomy, the portal vein was accessed to facilitate the administration of LEAD. Post-intervention, IVIS imaging of the rat liver that received the injection indicated a localized delivery of LEAD in the right medial lobe of the liver (Figures 61C and 61D). The cross-sectional IVIS images of treated and untreated lobes confirmed the delivery of LEAD into the target lobe (Figure 61E). Subsequent histological analysis showed successful tissue ablation with no viable cells and total loss of nuclei in the portal triad and periportal area in the LEAD group, in contrast to intact liver architecture and cellular composition in the untreated control group (Figures 61F and 61G). Immunohistochemistry studies showed that LEAD was able to deliver nivolumab effectively to the adjacent periportal areas. This aligns with the results from in vitro and ex vivo studies, confirming LEAD’s substantial drug delivery capabilities within the liver matrix (Figures 61F and 61G). Additionally, cytokeratin 7 (CK7) staining revealed the complete ablation of bile ducts with no cholangiocytes in the LEAD group, whereas in the control group, bile ducts and cholangiocytes remained intact (Figures 61F and 61G). Furthermore, CD31 immunostaining demonstrated significant ablation of endothelial cells in both the portal vein and hepatic arterioles in the LEAD group, in contrast to the control group (Figures 61F and 61G). In conclusion, the findings from the LEAD group indicated successful embolization and ablation of the portal triad and periportal area, along with marked ICG and nivolumab delivery into the liver parenchyma. Renal artery embolization using LEAD in a nonsurvival porcine model To further demonstrate LEAD’s capability for durable vascular embolization in a large animal model, a nonsurvival porcine kidney embolization model was selected due to its significant blood flow and a hierarchical vascular network (Krishnan et al., Mircocirculation, 23:e12661 (2021)). After achieving percutaneous access to the iliac artery, a 5F catheter was maneuvered through the aorta to reach the renal artery. Subsequently, 3 mL of LEAD was injected into the proximal main renal artery, successfully accomplishing embolization as evidenced by immediate post-embolization fluoroscopic imaging demonstrating the absence of any arterial flow into the kidney (Figures 76A and 76B). After one hour post embolization, the pigs were euthanized, and the kidneys were harvested for further imaging and analysis. Gross imaging of the kidneys revealed clear evidence of LEAD delivery with uniform detection of ICG using IVIS imaging (Figure 76C). Furthermore, on cross-sectional IVIS imaging, ICG was detected uniformly in the renal parenchyma, indicating the retention of LEAD in the parenchyma of the embolized kidneys (Figure 76D). Histological examinations further confirmed embolization in renal arterial branches (Figure 76E and 76F). This embolization was predominantly observed in arterioles with a diameter of 50 µm, accounting for 50.5% of cases, followed by those measuring 10 µm (29.9%) and 100 µm (12.8%) in diameter (Figure 76G), proving that LEAD can effectively reach and embolize even smaller vessels when injected into the larger upstream arteries with high blood flow. The ablation of renal arterial walls and adjacent areas is evident throughout histological sections (Figures 76F and 76H). This indicates that LEAD can diffuse even across the arterial walls, which are characteristically thicker in diameter and contain a greater amount of smooth muscle content compared to the walls of corresponding venous vessels (Wenceslau et al., Am J Physiol Heart Circ Physiol, 321:H77 (2021)), including the portal veins. Furthermore, it demonstrates LEAD’s ability to achieve successful tissue ablation even in high blood flow conditions, highlighting its effectiveness in penetrating and affecting tissues within these challenging high-flow environments. Immunohistochemistry staining confirmed the successful delivery of nivolumab within the embolized vessels and surrounding tissues (Figure 76F and 76I). HIF2α staining demonstrated hypoxia in the scattered parenchymal regions in the embolized kidneys compared to the untreated control kidneys, further confirming the successful embolization capabilities of LEAD (Figure 76F). These results indicate that LEAD is a potent embolic agent with remarkable ablation and drug delivery capabilities, particularly effective even in high blood flow systems such as the kidneys, where LEAD was able to reach vessels as small as 10 microns. Portal vein embolization with LEAD in a non-survival porcine liver model A porcine liver model was used to determine the feasibility of PVE using LEAD in a large animal model (Figure 77). Following percutaneous catheterization of the main portal vein under ultrasound guidance, a 5F Yueh catheter was advanced in the portal vein to the left lateral lobe of the porcine liver using and angled glidewire. Rapid and successful embolization of the portal vein was accomplished through the administration of 10 mL of LEAD via the 5F catheter, completed in less than a minute (Figures 62A – 62E). Figure 62C demonstrates in real-time injection of LEAD reaching the distal small branches of the portal vein; less than a minute after the delivery of LEAD, angiography in Figure 62D showed complete embolization with no flow into the embolized portal veins. This rapid embolization of the PVE is consistent with the rapid embolization observed in the porcine renal arteries in Figure 76. After the pigs were euthanized one-hour post-embolization, the livers were harvested for further imaging. The treated and untreated lobes of the porcine livers were cut axially. The ablation in the liver parenchyma was evident during the gross evaluation of the treated and untreated livers (Figure 62F). IVIS imaging of the liver sections confirmed the uniform delivery of ICG to the targeted lobe, with no ICG signal detected in the other lobes, signifying site-specific delivery and retention of LEAD in the embolized lobe of the liver (Figure 62F). This site-specific delivery was further confirmed in histological studies, indicating complete ablation of the portal triads and periportal parenchymal areas in the LEAD group, in marked contrast to the intact liver histology in the control group (Figures 62G and 62H). This targeted delivery was further confirmed by IHC staining, which revealed uniform distribution of nivolumab in the portal triad through periportal areas in the treated lobe, with no nivolumab signal in the control lobe (Figures 62G and 62H). Additionally, CK7 staining demonstrated successful ablation of cholangiocytes and bile ducts in the treatment group, whereas the control group displayed normal cholangiocyte cellularity (Figures 62G and 62H). CD31 staining also showed significant ablation of the endothelium in the portal veins and hepatic arteries of the treated lobe while remaining intact in the untreated lobes (Figures 62G and 62H). Furthermore, additional control portal vein embolization studies were conducted to demonstrate that IL3:1 is essential; in these control experiments, a solution composed of 70% PBS, 20% IOH, 1 mg / mL nivolumab, and 0.25 mg / mL ICG was delivered into the portal vein. IVIS imaging comparisons between treated and untreated lobes showed no difference in ICG signal, suggesting an absence of drug delivery or retention in the treated lobe (Figures 78A and 78B). Histological analysis of the liver sections revealed a normal liver with intact portal triads without any nivolumab signal in the treated lobe, emphasizing the vital role of IL3:1 in the LEAD formulation for embolization, ablation, and drug delivery capabilities (Figure 78C). Portal vein embolization with LEAD in a survival porcine liver model To evaluate the durability and long-term effects of PVE using LEAD, survival studies were conducted using a porcine liver model (Figure 79). Percutaneous access to the portal vein was achieved under ultrasound imaging guidance. Then, a 5F Yueh catheter was advanced to the left lateral lobe of the porcine liver under fluoroscopic imaging. Subsequently, 10 mL of LEAD was administered into the portal vein, resulting in successful embolization as evidenced by immediate post-embolization fluoroscopic imaging with iohexol (Figures 63A - 63D). Postoperatively, the pigs were observed for a period of seven days without any complications or signs of toxicity. Comparative blood analysis, including complete blood counts and serum marker tests, revealed no significant changes when pre- injection and one-week post-injection samples were compared (Table 4). Repeat fluoroscopic imaging on day 7 confirmed persistent embolization in the previously treated portal veins without any evidence for recanalization (Figure 63D). After euthanasia, the livers were harvested and sectioned in the axial plane for further imaging. Gross imaging of the treated lobe showed remarkable ablation. This finding was corroborated by IVIS imaging, which showed retention of ICG in the treated lobe, with no ICG signal in the untreated lobe. These findings indicate the persistent embolization of the portal veins and successful drug delivery by LEAD, with no adverse events such as recanalization or unintended embolization (Figures 63F and 63G). Histological analyses revealed complete ablation of portal triads extending to periportal parenchymal areas with no viable cells and complete loss of nuclei in the treated lobes (Figures 63G, 63H, 80A, and 80B). Immunohistochemistry studies showed that nivolumab was successfully delivered into the portal triad and adjacent parenchymal areas of the treated lobe, with no nivolumab signal detected in the untreated lobe (Figures 63G and 63H). CK7 staining indicated the absence of viable cholangiocytes or bile ducts in the treated lobe, in contrast to the intact bile ducts in the untreated lobe (Figures 63G and 63H). CD31 staining demonstrated complete ablation of the endothelium in the portal vein and hepatic artery within the portal triads of the treated lobe, while the endothelium in the untreated lobe's portal triads remained unaffected (Figures 63G and 63H). Additional immunohistochemistry staining, CD68 and MPO, was performed to evaluate any inflammatory response elicited by LEAD. Notably, both immunostainings yielded positive results in the densely cellular areas surrounding the ablation zone within the embolized lobe. This indicates a significant presence of recruited neutrophils and macrophages (Figure 80B). Moreover, Picrosirius red staining revealed fibrosis in the capsule and densely cellular regions, aligning with zones of immune cell infiltration in the embolized lobe. In contrast, the control group exhibited normal liver architecture (Figure 80B). These findings suggest that LEAD treatment can trigger immune cell activation and recruitment in the treated lobe, a process that is fundamental in cancer immunotherapy. Table 4. Comprehensive Analysis of porcine blood counts and serum chemistry. Murine cholangiocarcinoma tumor model To evaluate LEAD’s ablation and drug delivery capabilities within tumor matrices in an in vivo setting, a murine ectopic cholangiocarcinoma tumor model was established using the SB1 murine cholangiocarcinoma cell line (Figures 64A and 64B). LEAD was administered directly into the tumors under ultrasound guidance (Figure 64C), and the mice were subsequently euthanized one-hour post- injection. The harvested tumor tissues were then sectioned for further imaging analysis. IVIS imaging revealed a uniform distribution of ICG within the treated tumors (Figure 64D). Histological examination of the treated tumors showed successful tumor ablation across the histological sections (Figures 64E – 64G). Additionally, the uniform delivery of Nivolumab was confirmed through IHC staining (Figures 64E – 64G). These results indicate that LEAD is highly effective in inducing tumor ablation and facilitating drug delivery when directly injected into tumors. Evaluating the effect of LEAD on blood Comprehensive studies were conducted to evaluate the effect of LEAD on blood and its components, focusing on blood cells and coagulation dynamics. The hemolytic potential of LEAD was first assessed. It was found that LEAD concentrations up to 2% did not cause hemolysis, whereas concentrations at 5% and 10% resulted in complete hemolysis (Figures 81A and 81B). Peripheral blood smears supported these findings; intact red blood cells were observed with up to 2% LEAD concentration, but samples with 5% and 10% LEAD displayed cell aggregation and a lack of identifiable red blood cells (Figure 81C). While red blood cell counts in control groups and samples with up to 2% LEAD remained consistent, a significant decrease was noted at concentrations of 5% and 10% (Figure 82A). A similar pattern of cell reduction with increasing LEAD concentrations was observed in white blood cells, lymphocytes, monocytes, and granulocytes (Figures 82B – 82E), though platelet counts were relatively stable at lower concentrations (Figure 82F). Thrombosis assays demonstrated prolonged clotting times with higher concentrations of LEAD (Figure 83). Further, coagulation dynamics were assessed, revealing a significant prolongation of ACT as LEAD concentrations increased (Figure 84). Hemorheological evaluations indicated that higher LEAD concentrations led to a notable decrease in the G' value, indicative of alterations in the blood’s elastic properties (Figures 85A and 85B). These findings suggest that at dilute concentrations, LEAD leads to gelation of the blood, achieving rapid and successful embolization, with further interactions of LEAD with the vessel wall leading to durable occlusion. As the endothelial wall contains many pro-coagulation factors including von Willebrand factor (vWF) and tissue factor (Stojkovic et al., Scientific Reports, 6:25171 (2016); Yau et al., BMC Cardiovascular Disorders, 15:130 (2015)), the ablation of endothelium causes the release of these factors, and the interaction of these factors with blood components triggers the coagulation cascade, resulting in contributing to the rapid embolization visualized in animal studies. NON-LIMITING EMBODIMENTS The disclosure is illustrated herein by the following embodiments, which should not be construed as limiting. Those skilled in the art will understand that this disclosure may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. 1. A composition comprising an ionic liquid comprising geranic acid anions and choline cations, and a sugar, wherein the ratio of said geranate to said choline of said composition is from 6:1 to 1:6, optionally from 4:1 to 1:4, and further optionally from 3:1 to 1:3. 2. The composition of embodiment 1, wherein said composition comprises at least 30 percent, optionally at least 35 percent, of said ionic liquid. 3. The composition of embodiment 1, wherein said composition comprises at least 40 percent of said ionic liquid. 4. The composition of any one of embodiments 1-3, wherein said composition comprises least 20 percent, optionally at least 25 percent, of said sugar. 5. The composition of any one of embodiments 1-3, wherein said composition comprises from 25 to 35 percent of said sugar. 6. The composition of any one of embodiments 1-5, wherein the molar ratio of said geranic acid anions to said choline cations is from 1:1.5 to 1:6, optionally from 1:2 to 1:6. 7. The composition of any one of embodiments 1-5, wherein the molar ratio of said geranic acid anions to said choline cations is from 1:2.5 to 1:6. 8. The composition of any one of embodiments 1-5, wherein the molar ratio of said said geranic acid anions to said choline cations is from 1:2.5 to 1:5. 9. The composition of any one of embodiments 1-5, wherein the molar ratio of said geranic acid anions to said choline cations is (a) from 1:3 to 1:6, (b) from 1:3 to 1:5, (c) from 1:3 to 1:4, (d) from 1:3.5 to 1:6, (e) from 1:3.5 to 1:5, (f) from 1:3.5 to 1:4, or (g) from 1:3.5 to 1:4.5. 10. The composition of any one of embodiments 1-5, wherein the molar ratio of said geranic acid anions to said choline cations is 1:4. 11. The composition of any one of embodiments 1-10, wherein said composition comprises one or more imaging agents. 12. The composition of embodiment 11, wherein said composition comprises (a) from about 1 percent (v / v) to about 40 percent (v / v) of at least one of said one or more imaging agents, (b) from about 0.01 mg to about 500 mg of at least one of said one or more imaging agents per mL of said composition, (c) from about 0.05 mg to about 250 mg of at least one of said one or more imaging agents per mL of said composition, (d) from about 0.1 mg to about 100 mg of at least one of said one or more imaging agents per mL of said composition, or (e) from about 0.1 mg to about 10 mg of at least one of said one or more imaging agents per mL of said composition. 13. The composition of any one of embodiments 11-12, wherein (a) said composition comprises at least 1 percent of said one or more imaging agents, (b) said composition comprises at least 5 percent of said one or more imaging agents, (c) said composition comprises at least 10 percent of said one or more imaging agents, (d) said composition comprises at least 15 percent of said one or more imaging agents, (e) said composition comprises from about 1 percent to about 30 percent of said one or more imaging agents, (f) said composition comprises from about 5 percent to about 30 percent of said one or more imaging agents, (g) said composition comprises from about 10 percent to about 30 percent of said one or more imaging agents, or (h) said composition comprises from about 15 percent to about 25 percent of said one or more imaging agents. 14. The composition of any one of embodiments 11-13, wherein said composition comprises two or more imaging agents. 15. The composition of any one of embodiments 11-14, wherein said imaging agents are selected from the group consisting of iohexol, indocyanine green, and tantelum. 16. The composition of any one of embodiments 1-15, wherein said composition comprises one or more therapeutic agents. 17. The composition of embodiment 16, wherein said composition comprises (a) from about 1 mg to about 50 mg of at least one of said one or more therapeutic agents per mL of said composition, (b) from about 0.01 mg to about 500 mg of at least one of said one or more therapeutic agents per mL of said composition, (c) from about 0.05 mg to about 250 mg of at least one of said one or more therapeutic agents per mL of said composition, (d) from about 0.1 mg to about 100 mg of at least one of said one or more therapeutic agents per mL of said composition, or (e) from about 0.1 mg to about 10 mg of at least one of said one or more therapeutic agents per mL of said composition. 18. The composition of any one of embodiments 16-17, wherein (a) said composition comprises at least 1 percent of said one or more therapeutic agents, (b) said composition comprises at least 5 percent of said one or more therapeutic agents, (c) said composition comprises at least 10 percent of said one or more therapeutic agents, (d) said composition comprises at least 15 percent of said one or more therapeutic agents, (e) said composition comprises from about 1 percent to about 30 percent of said one or more therapeutic agents, (f) said composition comprises from about 5 percent to about 30 percent of said one or more therapeutic agents, (g) said composition comprises from about 10 percent to about 30 percent of said one or more therapeutic agents, or (h) said composition comprises from about 15 percent to about 25 percent of said one or more therapeutic agents. 19. The composition of any one of embodiments 16-18, wherein said composition comprises two or more therapeutic agents. 20. The composition of any one of embodiments 16-19, wherein a molecular weight of at least one of said one or more therapeutic agents is (a) greater than 7,500 Daltons, (b) greater than 10,000 Daltons, (c) greater than 15,000 Daltons, or (d) greater than 25,000 Daltons. 21. The composition of any one of embodiments 16-19, wherein a molecular weight of at least one of said one or more therapeutic agents is (a) greater than 50,000 Daltons, (b) greater than 100,000 Daltons, (c) greater than 125,000 Daltons, or (d) greater than 150,000 Daltons. 22. The composition of any one of embodiments 16-21, wherein a molecular weight of at least one of said one or more therapeutic agents is (a) less than 750,000 Daltons, (b) less than 500,000 Daltons, (c) less than 250,000 Daltons, or (d) less than 200,000 Daltons. 23. The composition of any one of embodiments 16-22, wherein a molecular weight of at least one of said one or more therapeutic agents is (a) from 7,500 Daltons to 750,000 Daltons, (b) from 10,000 Daltons to 500,000 Daltons, (c) from 15,000 Daltons to 250,000 Daltons, or (d) from 20,000 Daltons to 200,000 Daltons. 24. The composition of any one of embodiments 16-23, wherein at least one of said one or more therapeutic agents comprises a biologically active polypeptide. 25. The composition of embodiment 24, wherein said biologically active polypeptide is selected from the group consisting of nivolumab, ipilimumab, pembrolizumab, atezolizumab, durvalumab, cemiplimab, and avelumab. 26. The composition of any one of embodiments 16-25, wherein at least one of said one or more therapeutic agents comprises an immunogenic polypeptide that is immunogenic within a mammal. 27. The composition of embodiment 26, wherein said mammal is a human. 28. The composition of any one of embodiments 16-27, wherein at least one of said one or more therapeutic agents comprises an antigen-binding polypeptide. 29. The composition of embodiment 28, wherein said antigen-binding polypeptide is an antibody. 30. The composition of any one of embodiments 28-29, wherein said antigen-binding polypeptide is selected from the group consisting of nivolumab, ipilimumab, pembrolizumab, atezolizumab, durvalumab, cemiplimab, and avelumab. 31. The composition of any one of embodiments 16-30, wherein at least one of said one or more therapeutic agents has anti-cancer activity within a mammal. 32. The composition of embodiment 31, wherein said mammal is a human. 33. The composition of any one of embodiments 31-32, wherein said one or more therapeutic agents having anti-cancer activity are selected from the group consisting of doxorubicin and cisplatin. 34. The composition of any one of embodiments 1-33, wherein said composition comprises a nucleic acid. 35. The composition of embodiment 34, wherein said composition comprises (a) from about 5 mg to about 100 mg of said nucleic acid per mL of said composition, (b) from about 0.01 mg to about 500 mg of said nucleic acid per mL of said composition, (c) from about 0.05 mg to about 250 mg of said nucleic acid per mL of said composition, (d) from about 0.1 mg to about 100 mg of said nucleic acid per mL of said composition, or (e) from about 0.1 mg to about 10 mg of said nucleic acid per mL of said composition. 36. The composition of any one of embodiments 34-35, wherein (a) said composition comprises at least 1 percent of said nucleic acid, (b) said composition comprises at least 5 percent of said nucleic acid, (c) said composition comprises at least 10 percent of said nucleic acid, (d) said composition comprises at least 15 percent of said nucleic acid, (e) said composition comprises from about 1 percent to about 30 percent of said nucleic acid, (f) said composition comprises from about 5 percent to about 30 percent of said nucleic acid, (g) said composition comprises from about 10 percent to about 30 percent of said nucleic acid, or (h) said composition comprises from about 15 percent to about 25 percent of said nucleic acid. 37. The composition of any one of embodiments 34-36, wherein said nucleic acid encodes an imaging agent. 38. The composition of embodiment 37, wherein said imaging agent is selected from the group consisting of blue fluorescent protein (BFP), green fluorescent protein (GFP), and red fluorescent protein (RFP). 39. The composition of any one of embodiments 34-36, wherein said nucleic acid encodes a therapeutic agent. 40. The composition of embodiment 39, wherein said therapeutic agent encoded by said nucleic acid is selected from the group consisting of a biologically active polypeptide, an immunogenic polypeptide, and an antigen-binding polypeptide. 41. The composition of any one of embodiments 39-40, wherein said therapeutic agent encoded by said nucleic acid comprises anti-cancer activity within a mammal. 42. The composition of embodiment 41, wherein said mammal is a human. 43. The composition of any one of embodiments 1-42, wherein said composition has a viscosity of from about 15 millipascal second (mPa·s) to 35 mPa·s at 37°C. 44. The composition of any one of embodiments 1-42, wherein said composition has a viscosity of from about 20 millipascal second (mPa·s) to 30 mPa·s at 37°C. 45. The composition of any one of embodiments 1-44, wherein said composition is injectable into a blood vessel of a mammal. 46. The composition of embodiment 45, wherein said mammal is a human. 47. The composition of any one of embodiments 1-46, wherein said composition reduces blood flow within a blood vessel within a mammal when said composition is administered directly into the lumen of said blood vessel. 48. The composition of any one of embodiments 1-47, wherein said composition blocks blood flow within a blood vessel within a mammal when said composition is administered directly into the lumen of said blood vessel. 49. The composition of any one of embodiments 1-48, wherein tissue located outside of a blood vessel within a mammal and located within 20 cm of a blood occlusion site within said blood vessel is ablated when said composition is administered directly into the lumen of said blood vessel to form said blood occlusion site. 50. The composition of embodiment 49, wherein said tissue comprises cancer cells. 51. The composition of any one of embodiments 49-50, wherein said tissue is selected from the group consisting of kidney tissue, liver tissue, brain tissue, prostate tissue, pancreas tissue, breast tissue, lung tissue, colon tissue, and bladder tissue. 52. The composition of any one of embodiments 1-51, wherein said sugar is a sugar alcohol, a natural sugar, or a synthetic sugar; optionally, wherein said sugar is glycerol. 53. The composition of any one of embodiments 1-51, wherein said sugar is selected from the group consisting of dextrose, fructose, galactose, glucose, lactose, maltose, polysorbates, sucrose, xylose, erythritol, glycerol, hydrogenated starch hydrolysates, isomalt, lactitol, maltitol, mannitol, sorbitol, xylitol, agave, honey, and molasses. 54. The composition of any one of embodiments 1-53, wherein said composition is an embolic composition, optionally a catheter-directed embolic composition. 55. A method for treating a solid tumor within a mammal, wherein said method comprises administering a composition into a lumen of a blood vessel within said mammal, wherein said composition forms at least one blood occlusion within said blood vessel at a location within 20 cm of said solid tumor, wherein said composition comprises an ionic liquid comprising geranic acid anions and choline cations, and a sugar, wherein the ratio of said geranate to said choline of said composition is from 6:1 to 1:6, and optionally wherein at least some cancer cells of said solid tumor are ablated. 56. The method of embodiment 55, wherein said mammal is a human. 57. The method of any one of embodiments 55-56, wherein said solid tumor is a solid tumor located within a kidney, the liver, the brain, the prostate, the pancreas, a breast, a lung, the colon, or the bladder of said mammal. 58. The method of any one of embodiments 55-57, wherein said solid tumor comprises kidney cancer cells, liver cancer cells, brain cancer cells, prostate cancer cells, pancreatic cancer cells, breast cancer cells, lung cancer cells, colon cancer cells, or bladder cancer cells. 59. The method of any one of embodiments 55-58, wherein said composition comprises at least 0.1 percent of said ionic liquid, optionally at least 10 percent of said ionic liquid. 60. The method of any one of embodiments 55-58, wherein said composition comprises at least 25 percent of said ionic liquid. 61. The method of any one of embodiments 55-60, wherein said composition comprises at least 20 percent of said sugar, optionally at least 25 percent of said sugar. 62. The method of any one of embodiments 55-60, said composition comprises wherein about 28 percent of said sugar. 63. The method of any one of embodiments 55-62, wherein the molar ratio of said geranic acid anions to said choline cations is from 1:3.5 to 1:6. 64. The method of any one of embodiments 55-62, wherein the molar ratio of said geranic acid anions to said choline cations is from 1:3.5 to 1:5.5. 65. The method of any one of embodiments 55-62, wherein the molar ratio of said geranic acid anions to said choline cations is from 1:3.5 to 1:5. 66. The method of any one of embodiments 55-62, wherein the molar ratio of said geranic acid anions to said choline cations is from 1:3.5 to 1:4.5. 67. The method of any one of embodiments 55-62, wherein the molar ratio of said geranic acid anions to said choline cations is from 1:3.5 to 1:4. 68. The method of any one of embodiments 55-62, wherein the molar ratio of said geranic acid anions to said choline cations is 1:4. 69. The method of any one of embodiments 55-68, wherein said composition comprises an imaging agent. 70. The method of embodiment 69, wherein said composition comprises at least 10 percent of said imaging agent. 71. The method of any one of embodiments 69-70, wherein said imaging agent is selected from the group consisting of iohexol and tantelum. 72. The method of any one of embodiments 55-71, wherein said composition comprises a therapeutic agent. 73. The method of embodiment 72, wherein said composition comprises at least 0.01 percent of said therapeutic agent. 74. The method of any one of embodiments 72-73, wherein the molecular weight of said therapeutic agent is greater than 7,500 Daltons. 75. The method of any one of embodiments 72-73, wherein the molecular weight of said therapeutic agent is greater than 10,000 Daltons. 76. The method of any one of embodiments 72-73, wherein the molecular weight of said therapeutic agent is greater than 15,000 Daltons. 77. The method of any one of embodiments 72-73, wherein the molecular weight of said therapeutic agent is greater than 25,000 Daltons. 78. The method of any one of embodiments 72-73, wherein the molecular weight of said therapeutic agent is greater than 50,000 Daltons. 79. The method of any one of embodiments 72-73, wherein the molecular weight of said therapeutic agent is greater than 100,000 Daltons. 80. The method of any one of embodiments 72-73, wherein the molecular weight of said therapeutic agent is greater than 125,000 Daltons. 81. The method of any one of embodiments 72-80, wherein the molecular weight of said therapeutic agent is less than 750,000 Daltons. 82. The method of any one of embodiments 72-80, wherein the molecular weight of said therapeutic agent is less than 500,000 Daltons. 83. The method of any one of embodiments 72-80, wherein the molecular weight of said therapeutic agent is less than 250,000 Daltons. 84. The method of any one of embodiments 72-80, wherein the molecular weight of said therapeutic agent is less than 200,000 Daltons. 85. The method of any one of embodiments 72-84, wherein said therapeutic agent comprises a biologically active polypeptide. 86. The method of embodiment 85, wherein said biologically active polypeptide is selected from the group consisting of nivolumab, ipilimumab, pembrolizumab, atezolizumab, durvalumab, cemiplimab, and avelumab. 87. The method of any one of embodiments 72-86, wherein said therapeutic agent comprises an immunogenic polypeptide that is immunogenic within a mammal. 88. The method of embodiment 87, wherein said mammal is a human. 89. The method of any one of embodiments 72-84, wherein said therapeutic agent comprises an antigen-binding polypeptide. 90. The method of embodiment 89, wherein antigen-binding polypeptide is an antibody. 91. The method of embodiment 89, wherein said antigen-binding polypeptide is selected from the group consisting of nivolumab, ipilimumab, pembrolizumab, atezolizumab, durvalumab, cemiplimab, and avelumab. 92. The method of any one of embodiments 72-91, wherein said therapeutic agent has anti-cancer activity within said mammal. 93. The method of embodiment 92, wherein said therapeutic agent is selected from the group consisting of doxorubicin, cisplatin, paclitaxel, olaparib, everolimus, mitomycin, radioactive isotopes (e.g., yttrium Y-90, lutetium-177, actinium, fluorine-18, gallium-67, krypton-81m, rubidium-82, nitrogen- 13, technetium-99m, indium-111, iodine-123, xenon-133, and thallium-201), atezolizumab, bevacizumab, cabozantinib-s-malate, ramucirumab, pembrolizumab, lenvatinib mesylate, sorafenib tosylate, nivolumab, pemigatinib, pembrolizumab, ramucirumab, regorafenib, and abemaciclib. 94. The method of any one of embodiments 55-93, wherein said composition comprises a nucleic acid. 95. The method of embodiment 94, wherein said nucleic acid encodes an imaging agent. 96. The method of embodiment 95, wherein said imaging agent is selected from the group consisting of blue fluorescent protein (BFP), green fluorescent protein (GFP), and red fluorescent protein (RFP). 97. The method of embodiment 94, wherein said nucleic acid encodes a therapeutic agent. 98. The method of embod...

Claims

CLAIMS WHAT IS CLAIMED IS:

1. An embolic composition comprising an ionic liquid comprising geranic acid anions and choline cations, and a sugar, wherein the ratio of said geranate to said choline of said composition is from 6:1 to 1:6, optionally from 4:1 to 1:4, and further optionally from 3:1 to 1:

3.

2. The embolic composition of claim 1, wherein said composition comprises at least 30 wt% of said ionic liquid.

3. The embolic composition of claim 1, wherein said composition comprises at least 40 wt% of said ionic liquid.

4. The embolic composition of claim 1, wherein said composition comprises at least 20 wt% of said sugar.

5. The embolic composition of claim 1, wherein said composition comprises from 25 to 35 wt% of said sugar.

6. The composition of claim 1, wherein the molar ratio of said geranic acid anions to said choline cations is from 1:1.5 to 1:6, optionally from 1:2 to 1:

6.

7. The composition of claim 1, wherein the molar ratio of said said geranic acid anions to said choline cations is from 1:2.5 to 1:

5.

8. The composition claim 1, wherein the molar ratio of said geranic acid anions to said choline cations is 1:

4.

9. The composition of claim 1, wherein said composition comprises one or more imaging agents.

10. The composition of claim 9, wherein said imaging agents are selected from the group consisting of iohexol, indocyanine green, and tantelum.

11. The composition of claim 1, wherein said composition comprises one or more therapeutic agents.

12. The composition of any one of claims 11, wherein at least one of said one or more therapeutic agents comprises a biologically active polypeptide selected from the group consisting of nivolumab, ipilimumab, pembrolizumab, atezolizumab, durvalumab, cemiplimab, and avelumab.

13. The composition of claim 11, wherein said one or more therapeutic agents are selected from the group consisting of doxorubicin and cisplatin.

14. The composition of claim 1, wherein said composition comprises a nucleic acid that encodes an imaging agent selected from the group consisting of blue fluorescent protein (BFP), green fluorescent protein (GFP), and red fluorescent protein (RFP).

15. The composition of claim 1, wherein said composition comprises a nucleic acid encoding a therapeutic agent selected from the group consisting of a biologically active polypeptide, an immunogenic polypeptide, and an antigen-binding polypeptide.

16. The composition of claim 1, wherein said composition has a viscosity of from about 15 millipascal second (mPa·s) to 35 mPa·s at 37°C.

17. The composition of claim 1, wherein said composition is capable of administration through catheterization.

18. The composition of claim 1, wherein said sugar is selected from the group consisting of dextrose, fructose, galactose, glucose, lactose, maltose, polysorbates, sucrose, xylose, erythritol, glycerol, hydrogenated starch hydrolysates, isomalt, lactitol, maltitol, mannitol, sorbitol, xylitol, agave, honey, and molasses.

19. The composition of claim 1, wherein said sugar is glycerol.

20. A method for treating a solid tumor within a mammal, wherein said method comprises administering a composition of claim into a lumen of a blood vessel within said mammal, wherein said composition forms at least one blood occlusion within said blood vessel at a location within 20 cm of said solid tumor.

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