An aerogel vascular tumor model and uses thereof

Abstract

The application discloses an aerogel blood vessel tumor model and application thereof. The application utilizes a cell-attached tubular aerogel support and integrates the same into a perfusion culture device, and realizes construction of a three-dimensional model of a tumor invading a large blood vessel in a large size level (millimeter level) in vitro for the first time through a highly porous structure of the aerogel support and exchange of nutrients in a perfusion process, and overcomes technical difficulties in the prior art that the model cannot be constructed.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of medical simulation models, in particular to an aerogel blood vessel tumor model and application thereof. BACKGROUND

[0002] Liver cancer is one of the most common malignant tumors worldwide. Studies have shown that by 2025, the number of new liver cancer cases worldwide each year is expected to exceed 1 million. Among the many types of liver cancer, hepatocellular carcinoma (HCC) is the most common type of liver cancer, which mainly occurs in patients with chronic liver disease and cirrhosis. Due to the high invasiveness and early metastasis ability of HCC, its treatment is still facing great challenges. Vessel invasion is one of the most important factors for poor prognosis of HCC. Since HCC can cause cancer cells to invade blood vessels, especially portal veins and hepatic veins, it generally leads to intrahepatic spread and distant metastasis, therefore, vessel invasion caused by HCC often leads to postoperative recurrence and significant shortening of patient survival.

[0003] In the prior art, although mouse models can exhibit the vessel invasion characteristics of tumor cells, due to the small size of mouse tumors and blood vessels, as well as ethical restrictions, they have many deficiencies in fully replicating human diseases. Although traditional tumor organ chip models can simulate a tumor environment supported by microvessels, they still have deficiencies in reproducing tumor invasion of large blood vessels. Moreover, for general methods of constructing microvessel networks in vitro, due to the difficulty of removing metabolic products inside the tissue and the lack of sufficient nutrient supply, it is easy to cause necrosis inside the tissue, thereby greatly reducing the success rate of construction. Therefore, it can be seen that there are still great challenges in constructing large blood vessel channels and maintaining their structure and function in vitro.

[0004] Therefore, developing a HCC three-dimensional model with vessel invasion characteristics and allowing researchers to better observe the interaction between human liver cells and endothelial cells and the physiological response to drugs will be of great importance to researchers' understanding of HCC treatment. SUMMARY

[0005] The present application aims to solve at least one of the above-mentioned technical problems in the prior art. To this end, the purpose of the present application is to provide an aerogel blood vessel tumor model and application thereof. The present application utilizes a tubular aerogel scaffold for cell adhesion and integrates it into a perfusion culture device. Through the highly porous structure of the aerogel scaffold and the exchange of nutrients during the perfusion process, a three-dimensional model of tumor invasion of large blood vessels at a large size level (millimeter level) can be constructed in vitro.

[0006] In a first aspect, the present application provides an aerogel scaffold, wherein the raw material for preparing the aerogel scaffold comprises a lactide-caprolactone copolymer and 1,4-dioxane, and the mass-volume ratio of the lactide-caprolactone copolymer and 1,4-dioxane is 1-3:100.

[0007] In some embodiments of the present application, the mass-volume ratio of the lactide-caprolactone copolymer and 1,4-dioxane is 1.5-3:100.

[0008] In some embodiments of the present application, the mass-volume ratio of the lactide-caprolactone copolymer and 1,4-dioxane is 2:100.

[0009] In some embodiments of the present application, the raw material for preparing the aerogel scaffold only comprises a lactide-caprolactone copolymer and 1,4-dioxane.

[0010] In some embodiments of the present application, the aerogel scaffold is subjected to hydrophilic modification.

[0011] In some embodiments of the present application, the hydrophilic modification is achieved by plasma treatment. Of course, those skilled in the art can select other hydrophilic modification methods for treating the aerogel scaffold according to actual use requirements, including but not limited to plasma treatment, hydrophilic layer (coupling agent, surfactant or hydrophilic monomer or polymer) coating, ultraviolet irradiation, high-energy radiation and ozone method, etc.

[0012] In some embodiments of the present application, the aerogel scaffold is not limited in shape and can be designed into different tubular structures such as circular tube, square tube, rectangular tube, elliptical tube, hexagonal tube, etc.

[0013] In some embodiments of the present application, the aerogel scaffold has a porous structure, the pore size is 30-80 μm, the Young's modulus is 0.5-2 kPa, and the tensile modulus is 115-260%.

[0014] In a second aspect, the present application provides a preparation method of the aerogel scaffold of the above-mentioned aspect, comprising the following steps:

[0015] The mixed solution of the lactide-caprolactone copolymer and 1,4-dioxane is used to cast a mold, and the aerogel scaffold is obtained after freeze-drying.

[0016] In some embodiments of the present application, the mold can be prepared by any method in the art, including but not limited to 3D printing.

[0017] In some embodiments of the present application, the mold is obtained by 3D printing, and the 3D printing material is 8200pro resin material. Of course, those skilled in the art can also select other materials that facilitate demolding for printing according to actual use needs.

[0018] In some embodiments of the present application, the lactide-caprolactone copolymer and 1,4-dioxane are mixed and completely dissolved.

[0019] In some embodiments of the present application, the freeze-drying is performed by using a freeze dryer under vacuum conditions.

[0020] In some embodiments of the present application, the mixing and dissolving of the lactide-caprolactone copolymer and 1,4-dioxane are performed by stirring at a stirring rate of 1000 rpm for 2 hours or more, and then the mixed solution is introduced into the mold, frozen at-20℃ / -80℃ for 1 hour or more, and then the frozen mold is placed in a freeze dryer for overnight freeze-drying.

[0021] In some embodiments of the present application, the aerogel scaffold is subjected to hydrophilic modification after vacuum freeze-drying.

[0022] In some embodiments of the present application, the hydrophilic modification is achieved by plasma treatment. Of course, those skilled in the art can also select other hydrophilic modification methods for treating the aerogel scaffold according to actual use needs, including but not limited to plasma treatment, hydrophilic layer (coupling agent, surfactant or hydrophilic monomer or polymer) coating, ultraviolet irradiation, high-energy radiation and ozone method, etc.

[0023] In some embodiments of the present application, the hydrophilic modification is achieved by plasma treatment, and the plasma irradiation power is 120W and the irradiation time is 1min.

[0024] In a third aspect of the present application, the aerogel scaffold of the above-mentioned aspects is provided for use in preparing a medical simulation model.

[0025] In some embodiments of the present application, the medical simulation model is a blood vessel model.

[0026] In some embodiments of the present application, the medical simulation model is a tumor invasion blood vessel model.

[0027] In some embodiments of the present application, the medical simulation model is an HCC invasion blood vessel model.

[0028] In a fourth aspect of the present application, a vascular tumor simulation model is provided, which comprises the aerogel scaffold of the above aspect, endothelial cells attached to the inner surface of the aerogel scaffold, and tumor cells attached to the outer surface of the aerogel scaffold.

[0029] In some embodiments of the present application, the endothelial cells and the tumor cells are not attached to the same place.

[0030] In a fifth aspect of the present application, a method for constructing a vascular tumor simulation model is provided, which comprises the following steps:

[0031] The endothelial cells and the tumor cells are respectively seeded on the inner surface and the outer surface of the aerogel scaffold of the above aspect, and after the cells are fixed on the aerogel scaffold, perfusion culture is performed, and after 1-6 days of culture, a vascular tumor simulation model is obtained.

[0032] In some embodiments of the present application, the aerogel scaffold is continuously rotated during seeding.

[0033] In some embodiments of the present application, the cell concentration of the endothelial cells and the tumor cells used for seeding is 3-5 x 10 7 / mL.

[0034] Of course, the cell concentration of the endothelial cells and the tumor cells used for seeding can be reasonably adjusted by the skilled person in the art based on the actual situation.

[0035] In some embodiments of the present application, the endothelial cells can include HUVEC, and the tumor cells can include hepatocarcinoma cells, such as Hep3B cells. Of course, the skilled person in the art should understand that the tumor invasion of large vessels described in the present application is not limited to hepatocarcinoma, and other types of tumors can also have such phenomenon, such as pancreatic cancer, renal cell carcinoma and lung cancer, and therefore, the simulation model in the present application can also be used to simulate the tumor invasion of large vessels caused by other tumors.

[0036] In a sixth aspect of the present application, the vascular tumor simulation model of the above aspect or the vascular tumor simulation model obtained by the construction method of the above aspect is provided for use in an in vitro disease simulation model.

[0037] In some embodiments of the present application, the disease includes a tumor.

[0038] In some embodiments of the present application, the disease is a vascular invasive tumor.

[0039] In some embodiments of the present application, the disease is HCC.

[0040] In a seventh aspect, the application provides use of the blood vessel tumor simulation model according to any of the above aspects or the blood vessel tumor simulation model obtained by the construction method according to any of the above aspects in constructing an in-vitro drug screening platform.

[0041] The application has the following beneficial effects:

[0042] 1. The application first proposes a tumor invasion large blood vessel simulation model constructed by using a cell-attached tubular aerogel scaffold, which solves the blank of no in-vitro model in the prior art. Meanwhile, compared with a conventional mouse tumor model, the model can realize construction of a large blood vessel and a large-size tumor, while the conventional mouse tumor model is difficult to effectively simulate the process due to the fact that a mouse tumor and a blood vessel cannot grow to this size level and due to ethical restrictions.

[0043] 2. The model construction method in the application is simple and fast, can be easily implemented, and realizes satisfaction of various requirements such as a highly porous structure and exchange of nutrients by using the tubular aerogel scaffold in combination with a perfusion culture device, thereby overcoming the technical defects that cannot be constructed in the prior art and realizing the technical effect of constructing a three-dimensional model of a large-size level (millimeter level) in vitro. BRIEF DESCRIPTION OF DRAWINGS

[0044] Figure 1 A complete flowchart for preparing the aerogel blood vessel tumor model.

[0045] Figure 2 A physical diagram of the 3D-printed scaffold mold.

[0046] Figure 3 Changes in hydrophilicity and hydrophobicity of the scaffold surface before and after plasma treatment.

[0047] Figure 4 A physical diagram of the tubular aerogel scaffold.

[0048] Figure 5 Scanning electron microscope images of Examples 1-4.

[0049] Figure 6 A tensile diagram of the tubular aerogel scaffold.

[0050] Figure 7 A schematic diagram of the perfusion device.

[0051] Figure 8 Infiltration of cells on the scaffold after perfusion for one day (DAPI is used for cell nucleus staining, and Cy5 is used for scaffold staining) (the scale is 200 μm).

[0052] Figure 9Images showing the distribution of HUVEC and Hep3B cells at the front, middle, and rear ends of the scaffold (scale bar 500 μm).

[0053] Figure 10 Immunostaining images of VE-Cadherin and CD31 on the scaffold (scale bar 200 μm).

[0054] Figure 11 Image showing the cell distribution when only Hep3B cells are on the scaffold (scale bar 500 μm).

[0055] Figure 12 Images of Ki67 staining on cells on scaffolds under three culture conditions (scale bar 200 μm).

[0056] Figure 13 Live / dead staining images of the scaffold after co-culturing with three drugs (sorafenib, lenvatinib, and LNP-PLK1) for 48 h (scale bar 200 μm).

[0057] Figure 14 The image shows Ki67 immunofluorescence staining after the stent was co-cultured with three drugs (sorafenib, lenvatinib, and LNP-PLK1) for 48 h (scale bar: 200 μm). Detailed Implementation

[0058] The present invention will be further described in detail below through specific embodiments. Unless otherwise specified, the raw materials, reagents, or apparatus used in the embodiments and comparative examples are all available from conventional commercial sources or can be obtained by existing technical methods. Unless otherwise specified, the test or experimental methods are conventional methods in the art.

[0059] Example 1

[0060] This embodiment provides a method for constructing an aerogel vascular tumor model, the process of which is as follows: Figure 1 As shown, the specific steps include the following:

[0061] (1) Construction of aerogel scaffold:

[0062] A mold for fabricating the tubular scaffold was designed using 3D printing to meet the required size. Then, lactide-caprolactone copolymer (PLCL) powder was fully added to a 1,4-dioxane solution and stirred at 1000 rpm for at least 2 hours at room temperature until fully dissolved, forming a mixed solution. After removing air bubbles, the solution was poured into the prepared tubular scaffold mold. The mold was then frozen at -20℃ / -80℃ for at least 1 hour until completely frozen, and then transferred to a freeze dryer for overnight freeze-drying to obtain a tubular aerogel scaffold with a porous structure.

[0063] In the 3D printing, the mold design for the tubular stent can be achieved using SolidWorks. The 3D printing material is selected as 8200pro resin material.

[0064] In the present embodiment, the mass-volume ratio of the PLCL powder to the 1,4-dioxane solution is 1.5:100.

[0065] The obtained mold of the tubular stent is shown in Figure 2 .

[0066] The obtained tubular aerogel stent with a porous structure is a super-hydrophobic material per se. In order to facilitate the adhesion and growth of cells, the obtained tubular aerogel stent is placed in a vacuum plasma pump, and is treated for 1 minute using a power of 120 W, so as to complete the modification treatment of the surface of the tubular aerogel stent. After the modification treatment, the tubular aerogel stent is converted into a super-hydrophilic material, so as to facilitate the adhesion of cells and the penetration of culture medium (as shown in Figure 3 .

[0067] (2) Construction of a liver cancer model:

[0068] 50 μL of 5 x 10 7 / mL human liver cancer cell line Hep3B and 30 μL of 5 x 10 7 / mL human umbilical vein endothelial cells (HUVEC) are respectively planted on the outer wall surface and the inner wall surface of the tubular aerogel stent obtained in step (1), and are first placed in a 37°C cell incubator for overnight culture, and then are connected to a peristaltic pump for perfusion culture. The culture method is perfusion culture. In a 37°C, 5% CO2 cell incubator, the culture is performed for 5 days using DMEM culture medium: ECM culture medium = 1:1 (volume ratio), so as to form a dense tumor tissue. Based on the difference in the planted cells, a layer of endothelial cell barrier is formed on the inner wall of the tubular aerogel stent, and the outer wall is tightly surrounded by liver cancer cells, so as to form a vascular invasion structure, and an aerogel vascular tumor model is obtained.

[0069] The tubular aerogel stent with a porous structure in step (1) is shown in Figure 4 .

[0070] Example 2

[0071] The present embodiment provides a construction method of an aerogel vascular tumor model, and the flow thereof is the same as that of Example 1, except that the mass-volume ratio of the PLCL powder to the 1,4-dioxane solution is 2:1.

[0072] Example 3

[0073] The embodiment provides a method for constructing an aerogel hemangioma model, and the procedure is the same as that in Embodiment 1, except that the mass-volume ratio of the PLCL powder and the 1,4-dioxane solution is 2.5:100.

[0074] Embodiment 4

[0075] The embodiment provides a method for constructing an aerogel hemangioma model, and the procedure is the same as that in Embodiment 1, except that the mass-volume ratio of the PLCL powder and the 1,4-dioxane solution is 3:100.

[0076] Embodiment 5

[0077] The surface and cross-section pore size of the aerogel hemangioma models prepared in Embodiments 1-4 are characterized by a scanning electron microscope (SU8220, Japan Hitachi) under a 10kV accelerating voltage.

[0078] The results are shown in Figure 5 .

[0079] It can be found that the aerogel hemangioma models prepared in Embodiments 1-4 all have a uniform porous structure, which is beneficial to the connection and exchange between cells. In addition, with the increase of the PLCL content, the pore size of the tubular aerogel scaffold with a porous structure gradually decreases.

[0080] Embodiment 6

[0081] The tubular scaffold is prepared according to the method in Embodiments 1-4 by using a mold with a length of 25mm, a width of 10mm and a thickness of 3mm, and then a tensile test is performed (as shown in Figure 6 The tensile test is performed by using a general test system (Instron 68TM-5, USA), and the travel speed of the measurement is 0.5mm min -1 .

[0082] The Young's modulus and the tensile modulus of the tubular scaffold are shown in Table 1.

[0083] Table 1 Young's modulus and tensile modulus of the tubular scaffold

[0084] Item Example 1 Example 2 Example 3 Example 4 Pore size (pm) 77.75 68.20 45.31 35.83 Young's modulus (kPa) 0.548 0.889 1.564 1.880 Tensile modulus (%) 118.98 250.82 189.44 183.78

[0085] It can be found that with the increase of the PLCL content, the Young's modulus of the material also increases, and the aerogel material in Embodiment 2 has the best ductility.

[0086] Embodiment 7

[0087] In this embodiment, when preparing according to the methods in Examples 1-4, after obtaining the mixed solution of PLCL and 1,4-dioxane in step (1), Cy5 with a final concentration of 10 μg / mL was added. After completing the subsequent steps, a tubular aerogel scaffold with Cy5 fluorescence was obtained. The cell density was 5 × 10⁻⁶ cells / mL. 7 HUVEC cell suspension of 10 cells / mL and 5×10 7 Hep3B cell suspensions were mixed at a 1:1 ratio, and 50 μL of the mixture was applied to the outer surface of a tubular aerogel scaffold, while 30 μL was applied to the inner surface. The scaffold was then incubated for 40 min. During this time, culture medium (DMEM:ECM = 1:1) was continuously added to maintain cell viability. Finally, sufficient culture medium was added and the scaffold was incubated overnight. The culture was performed using perfusion culture, and the perfusion culture apparatus used was as follows: Figure 7 As shown.

[0088] After model construction, the model was fixed with 4% paraformaldehyde for 2-24 hours, then placed in an OCT complex (Sakura Finetek Japan Co., Ltd., Tokyo, Japan) and frozen to obtain 100 μm sections. Cell infiltration was observed under a confocal microscope (Nikon, A1R, Japan).

[0089] The results are as follows Figure 8 As shown.

[0090] It can be observed that after one day of perfusion, the models in Examples 1 and 2 achieved complete cell infiltration, while in Examples 3 and 4, due to the pore size, only partial cell infiltration occurred, and incomplete infiltration was observed. Furthermore, considering Young's modulus and extensibility, the scaffold fabricated in Example 2 was the most advantageous for subsequent liver cancer model construction among the four examples. However, for the other examples, based on the differences in the cells used and the duration of perfusion, they can be used for the seeding and model construction of other cells.

[0091] Example 8

[0092] The scaffold in Example 2 was processed using the method described in the above embodiments. In this case, human hepatocellular carcinoma cell line Hep3B and human umbilical vein endothelial cells HUVEC were seeded on the outer and inner surfaces of the tubular scaffold, respectively, and then perfused and cultured for five days to form dense tumor tissue. An endothelial barrier could be formed inside the scaffold, and the outside of the scaffold was tightly surrounded by hepatocellular carcinoma cells, forming a vascular invasion structure.

[0093] The specific steps are as follows:

[0094] (1) In order to facilitate the observation of the position distribution of HUVEC and Hep3B cells, 2 hours before the digestion of the two kinds of cells, DiO and DiD membrane dyes were diluted at a ratio of 1:1000 and added to the corresponding cell culture medium to label the two kinds of cells. Then, after the HUVEC and Hep3B cells were digested using 0.25% trypsin, the cells were collected using a 1.5 mL centrifuge tube and placed on ice, wherein the cell density was controlled to be 5x10 7 / tube.

[0095] (2) When the cell seeding of the stent is performed, the stent is continuously rotated. Specifically, under the condition of rotating the stent, 50 μL of Hep3B cell suspension is first added to the outside of the stent, and then 30 μL of HUVEC cell suspension is added to the inside. The stent with completed cell seeding is placed in a cell culture incubator at 37°C for 40 minutes, during which the culture medium is continuously taken out and supplemented.

[0096] (3) After overnight culture in the cell culture incubator, the stent is perfusion cultured using a peristaltic pump, wherein the culture medium is a mixed culture medium of DMEM and ECM (V:V=1:1).

[0097] (4) After five days of perfusion culture, the stent is taken out, and the front, middle and rear sections are frozen sectioned and observed using a confocal microscope.

[0098] The results are shown in Figure 9 .

[0099] It can be found that the three section images of the front, middle and rear show the uniformity of the cell distribution inside the stent, and the HUVEC cells form an endothelial layer-like structure inside the stent, while the Hep3B cells are closely surrounded outside the endothelial layer, together forming a vascular invasion structure.

[0100] VE-Cadherin and CD31 are markers for indicating angiogenesis and tight junction of endothelial cells. By further immunostaining the sections using VE-Cadherin and CD31, it can be found that the endothelial cells can form a tight cell junction on the inner wall of the stent, form a vascular barrier and promote the generation of microvessels Figure 10 .

[0101] In order to further verify the effectiveness of the stent in modeling, the inventors also tested the case of using only Hep3B cells for seeding according to the above method. It was found that when only Hep3B cells were seeded on the stent and perfusion cultured, only cells were found on the outside of the stent, and no cells were detected inside the stent Figure 11 . This indicates that when Hep3B cells exist alone, the cells can only grow outside the stent and cannot penetrate into the stent.

[0102] Ki67 is a positive marker of cell proliferation. The inventors designed three different culture conditions (co-culturing of HUVEC cells and Hep3B cells on perfusion-cultured scaffolds; only Hep3B cells on perfusion-cultured scaffolds; co-culturing of HUVEC cells and Hep3B cells on static-cultured scaffolds) and then performed section staining on the scaffolds.

[0103] The results are shown in Figure 12

[0104] Under perfusion culture condition, when Hep3B and HUVEC were co-cultured on the scaffolds, cells on the edge and inside of the scaffolds showed strong cell proliferation activity. When only Hep3B cells were present, cells on the edge of the scaffolds also continued to proliferate. However, when Hep3B and HUVEC were co-cultured statically, cells could not enter the scaffolds and the cell proliferation activity was low. This indicates that perfusion culture can continuously supply nutrients to cells on the scaffolds and remove metabolic waste to promote cell proliferation, while static culture limits metabolic activity and material exchange between cells.

[0105] Example 9

[0106] Sorafenib and Lenvatinib are currently the most effective single-agent therapies for patients with advanced hepatocellular carcinoma, while lipid nanoparticles (LNPs) based on siRNA are emerging drugs that show broad application prospects in the field of cancer treatment. Therefore, by comparing the effects of sorafenib, lenvatinib and LNPs encapsulating siRNA on the model, the predictive ability of the model for drug treatment effectiveness can be verified.

[0107] Polo-like kinase 1 (PLK1) protein is highly expressed in various cancers and plays a key role in cell cycle control and DNA damage response, and is one of the molecules necessary for rapid proliferation of tumor cells. In addition, PLK1 is also significantly highly expressed in patients with hepatocellular carcinoma with poor prognosis. Therefore, siRNA-PLK1 encapsulated by LNPs was compared with sorafenib and lenvatinib to verify the ability of the model in drug screening.

[0108] wherein the siRNA-PLK1 sense strand is 5'-UGAAGAAGAUCACCCUCCUUAdTdT-3' (SEQ ID NO: 1) and the antisense strand is 5'-UAAGGAGGGUGAUCUUCUUCAdTdT-3' (SEQ ID NO: 2). Wherein d represents deoxyribonucleotide.

[0109] ​The model after perfusion culture for five days was co-cultured with 2.5 μg / mL sorafenib, 2.5 μg / mL lenvatinib and 150 nM LNP-PLK1 respectively for 48 hours, and then the cells in the model were subjected to live / dead staining analysis.

[0110] The results are shown in Figure 13

[0111] It can be found that the cells in the three experimental groups all have significant death phenomenon (red fluorescence).

[0112] Since these drugs mainly inhibit disease progression by inhibiting cell proliferation, the inventors further compared the changes in cell proliferation after drug treatment, wherein Ki67 immunofluorescence staining in the above example was used to characterize the changes in cell proliferation.

[0113] The results are shown in Figure 14

[0114] The results show that the three drugs all have a certain inhibitory effect, and the inhibitory effect of sorafenib and lenvatinib is more significant. The inventors believe that this difference may be due to various factors, for example, sorafenib and lenvatinib as multi-target kinase inhibitors can directly block multiple cancer-related signaling pathways, and the stability of the lipid nanoparticles is low.

[0115] ​​The prior art (El Dika, I, et al. "An Open-Label, Multicenter, Phase I, Dose Escalation Study with Phase II Expansion Cohort to Determine the Safety, Pharmacokinetics, and Preliminary Antitumor Activity of Intravenous TKM-080301 in Subjects with Advanced Hepatocellular Carcinoma." The Oncologist 24.6 (2019): theoncologist.2018-0838-.) confirms that si-PLK1 has obvious inhibitory effect on cell proliferation on two-dimensional cultured cells and in a mouse subcutaneous tumor model, but the anti-tumor effect in clinical human trials is limited, and there is no significant tumor inhibition effect, thereby verifying the accuracy of the above conclusion. It can be shown that, compared with the traditional mouse model, the model in the present application can serve as a more effective, more accurate and more convenient drug evaluation platform closer to the clinical results for screening and evaluation of different types of drugs, and has high application potential.

[0116] The above embodiments are preferred embodiments of the present application, but the embodiments of the present application are not limited by the above embodiments, and any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present application shall be equivalent replacement methods and shall be included in the protection scope of the present application.

Claims

1. A simulation model for vascular tumors, characterized in that, The vascular tumor simulation model includes an aerogel scaffold, endothelial cells attached to the inner surface of the aerogel scaffold, and tumor cells attached to the outer surface of the aerogel scaffold. The aerogel scaffold is prepared using lactide-caprolactone copolymer and 1,4-dioxane, with a mass-to-volume ratio of (1-3):

100. The aerogel scaffold has been modified to be hydrophilic; The aerogel scaffold is a porous tubular aerogel scaffold.

2. The vascular tumor simulation model according to claim 1, characterized in that, The mass-to-volume ratio of the lactide-caprolactone copolymer and 1,4-dioxane is (1.5-3):

100.

3. A method for constructing a vascular tumor simulation model, comprising the following steps: Endothelial cells and tumor cells are seeded on the inner and outer surfaces of the aerogel scaffold described in claim 1 or 2, respectively. After the cells are fixed to the aerogel scaffold, they are perfused and cultured for 1-6 days to obtain a vascular tumor simulation model. During planting, the aerogel support is rotated continuously.

4. The construction method according to claim 3, characterized in that, The cell concentration of endothelial cells and tumor cells used for implantation was (3-5)×10⁻⁶. 7 / mL.

5. The application of the vascular tumor simulation model according to claim 1 or 2 or the vascular tumor simulation model obtained by the construction method according to any one of claims 3-4 in an in vitro disease simulation model, wherein the disease includes tumors.

6. The application of the vascular tumor simulation model according to claim 1 or 2 or the vascular tumor simulation model obtained by the construction method according to any one of claims 3-4 in the construction of an in vitro drug screening platform.

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