Hybrid bioelectronic devices and methods of manufacture and use thereof

Abstract

Nanoelectronic devices and bioelectronic hybrids thereof are described herein. Methods of making and using such devices and hybrids are also described. The nanoelectronic devices and bioelectronic hybrids can be used, for example, in medical applications. For example, the bioelectronic hybrids can be non-surgically implanted to provide stimulation or modulation to a target site or region in a subject.

Description

[0001] HYBRID BIOELECTRONIC DEVICES AND METHODS OF MANUFACTURE AND USE THEREOF

[0002] CROSS-REFERENCE TO REEATED APPLICATIONS

[0003] This application claims priority to U.S. Provisional Patent Application No. 63 / 557,271, filed February 23, 2024, which is incorporated herein by reference.

[0004] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0005] This invention was made with government support under HL 168072 awarded by the National Institutes of Health. The government has certain rights in the invention.

[0006] FIELD OF THE INVENTION

[0007] This invention is in the field of nanoelectronic devices and bioelectronic hybrids thereof which can be implanted into patients and can be used for various medical applications.

[0008] BACKGROUND OF THE INVENTION

[0009] Electronics leverage the last five decades of growth and development in information technology, with integration of electronic, mechanical, optical and magnetic systems (Shalf, J. The future of computing beyond Moore’s Law. Philos. Trans. R. Soc. A 378, 20190061 (2020)). Such devices when implanted in the body, can provide powerful tools for diagnosis, therapeutics and fundamental research (Won. S., el al., Cell 181. 115-135 (2020); Heck, C. Crit. Care Nurs. Clin. 28, 77-86 (2016); Verrills. P., et al., J. Pain Res. 9, 481-492 (2016); and Beck. H. et al., Am. J. Cardiol. 106, 810-818 (2010)). However, despite the progress several limitations and drawbacks exist with current devices.

[0010] For example, bioelectronic implants for brain stimulation have provided fundamental biological insights as well as have been proven to be effective for treating many brain diseases (Won, S., et al.. Cell 181, 115-135 (2020)). However, placing a medical implant (be it wired or wireless) inside the brain typically requires disruption of the skull and invasive intracranial surgery, which is associated with pain, tissue damage, infection along with risks of ischemia, psychological distress, morbidity and mortality (H. Richard Winn, et al., Youmans Winn Neurol. Surg. (2017)). Furthermore, intracranial surgery can disrupt neural networks and constrain fundamental studies. Even endovascular electrodes (Fan, J. Z., et al., Frontiers in Neuroscience 14, 432 (2020); Zhang, A. et al., Science 381, 306-312 (2023); Opie, N. L. et al. Nat. Biomed. Eng. 2, 907-914 (2018), though not requiring intracranial access, still need surgical implantation (endovascular) with its associated risks and complications. Moreover, the aforementioned implants and electrodes cannot achieve sub-millimeter spatial targeting precision and in addition, are unable to access most of the brain regions.

[0011] While attempts have been made to explore intravenous injection route (Nguyen, T. el al., Neurotherapeutics 18, 2091-2106 (2021)), they have led to non-specific stimulation of large brain regions without focality. On the other hand, existing non-invasive brain stimulation technologies (these do not involve any implants inside the brain) such as Transcranial Magnetic Stimulation (TMS) and Transcranial Direct Current Stimulation (TDCS), lack necessary spatiotemporal resolution (Rossini, P. M. etal., Clinical Neurophysiology’ 126, 1071-1107 (2015)).

[0012] Beyond the issues identified above relating to devices for the brain, the same or similar limitations and drawbacks exist in using such devices in other organs and regions of the body. Thus, there remains a need for devices which can be implanted in different organs and regions of the body non-surgically and which can address the shortcomings of currently known devices.

[0013] SUMMARY

[0014] In one aspect, a hybrid bioelectronic device is provided. In some particular embodiments, the hybrid bioelectronic device comprises a wirelessly actuatable component configured to interact with an externally applied stimulus; and a biological or chemical component attached to the wirelessly actuatable component, wherein the hybrid bioelectronic device is configured to be administered into a body of a subject and thereafter recognize, via the biological or chemical component, a target region within the body and self-implant at the target region.

[0015] In another aspect, a method of making a hybrid bioelectronic device is provided. In some particular embodiments, the method comprises: manufacturing, via top-down microfabrication techniques or bottom-up chemical synthesis, a wirelessly actuatable component configured to interact with an externally applied stimulus, wherein the wirelessly actuatable component is substrate-free; and attaching a biological or chemical component to the wirelessly actuatable component to produce the hybrid bioelectronic device, wherein the hybrid bioelectronic device is configured to be administered into a body of a subject and thereafter recognize, via the biological or chemical component, a target region within the body and selfimplant at the target region.

[0016] In still another aspect, a method of use is provided. In some particular embodiments, the method comprises: introducing one or more hybrid bioelectronic devices into a body of a subject in need thereof; permitting the one or more hybrid bioelectronic devices to travel within the body of the subject to a target region and to self-implant at the target region; and applying a stimulus to a wirelessly actuatable component of the self-implanted one or more hybrid bioelectronic devices which are configured to interact with the stimulus

[0017] BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1A shows a non-limiting representation of nanoelectronic device 100, which works on the photovoltaic principle, and is a three-layered device based on organic polymers where layer 110 is an anode, layer 120 is an organic semiconductor, and layer 130 is cathode.

[0019] Figure IB shows a scanning electron microscopy (SEM) images of a nanoelectronic device fabricated at wafer scale. Left image scale bar - 10 pm and right image scale bar - 1 pm.

[0020] Figure 1C is a 3D-reconstructed FIB-SEM image of a nanoelectronic device attached to an immune cell.

[0021] Figure 2A shows a schematic diagram of a setup used for in-air and ex-vivo measurements, where nanoelectronic devices are illuminated from the bottom with a laser. The microscope is used to assist with alignment while probing the nanoelectronic devices from the top using the 3 -stage micromanipulators. The probes are connected to a potentiostat, and the laser is operated in continuous wave (CW) mode to obtain the current-voltage (IV) curves.

[0022] Figure 2B is a representative plot showing the pow er generated by different-sized nanoelectronic devices as a function of applied voltage. Nanoelectronic device structure - PEDOT:PSS|P3HT:PCBM|Ti. Intensity - 10 mW / mm2incident on the devices. Legend - device diameter.

[0023] Figure 2C is a representative plot showing the IV characteristics of a nanoelectronic device (10 pm in diameter) for varied light intensities incident on the devices. Nanoelectronic device structure - PEDOT:PSS|P3HT:PCBM|Ti. A 520 nm wavelength laser was used as the light source. Legend - incident light intensities.

[0024] Figure 2D is a representative IV plot for the nanoelectronic devices (10 pm in diameter) with light passing through different thicknesses of brain slices or whole brain. Nanoelectronic device structure -PEDOT:PSS|PCPDTBT:PCBM |Ti. Laser - 792 nm, intensity - 24.6 mW / mm2incident on the bottom surface of the brain.

[0025] Figure 2E is a representative plot of the maximum power generated by the nanoelectronic devices (10 pm in diameter) with light passing through the whole brain without the skull and the whole brain with an intact skull at different incident light intensities (792 nm) incident on the bottom surface of the brain. Nanoelectronic device structure - PEDOT:PSS|PCPDTBT:PCBM|Ti, values represent median ± standard deviation (SD). Figure 3 A shows a histogram of the population of nanoelectronic devices, as bioelectronic hybrids with an immune cell, and only cells in the suspension prepared for intravenous injections before and after fluorescence-activated cell sorting (FACS). A double gating strategy was employed to isolate the bioelectronic hybrids from the suspension with a purity of 92.4% ± 5.2% (n>5).

[0026] Figure 3B shows a bar graph of the titanium content in the brain (for quantification of the self-implanted nanoelectronic devices, as devices alone and as bioelectronic hybrids thereof with an immune cell, which contain a titanium layer compared with the intrinsic baseline titanium content in mouse brain) measured by Inductively Coupled Plasma Mass Spectrometry (ICP-MS) when 2 x 106 number of devices only or hybrids were injected into the animal intravenously for the experimental and control groups. A significant increase in titanium content compared to the baseline was observed only for the experimental group while for both the control groups, the changes compared to baseline were insignificant, values represent mean ± SD (n = 3), ***p<0.001 (one-way ANOVA test).

[0027] Figure 3C shows a bar graph of the coefficient (01 ) from the logistic regression model suggesting an influence of inflammation on localization of the hybrids in the actual experimental data (01 =2.2), compared to the shuffled control (01=10-4); **p<0.01 (student t-test).

[0028] Figure 3D shows a bar graph of the accuracy (ACC) values which indicated a better model fit for the experiment (ACC=0.82±0.05) compared to the shuffled control (ACC=0.50±0.01). supporting the assumption of hybrid localization to areas of inflammation (high LPS intensity); ****p<0.0001 (student t-test).

[0029] Figure 4A shows a schematic diagram of the timeline and the wireless stimulation process of the bioelectronic hybrids of the nanoelectronic devices attached to an immune cell. For the experimental group optical actuation was applied (n = 4 mice) and for the control group no optical actuation was applied (n = 5 mice). The optical pulse sequence applied was 792 nm, 15 mW / mm2, 10 ms pulse width, 20 Hz, 20 minutes duration. Nanoelectronic device structure - PEDOT:PSS PCPDTBT:PCBM|Ti|TiN.

[0030] Figure 4B shows a Scatter plot of the number of c-Fos positive cells in the target (inflamed) region for the experimental (with wireless optical actuation) and control (without actuation) groups. Longer (shorter) horizontal lines represent the median (variance) of the data; **p- value = 0.0017 (student t-test).

[0031] Figure 4C shows a bar plot showing the distribution of c-Fos positive cells as a function of radial distance from the inflammation boundary (represented by 0 in the x-axis) for the experimental and control groups, values represent mean ± SEM; *p<0.05 (student t-test). Figure 5 shows two energy band diagrams for PEDOT:PSS|P3HT: PCBM|Ti (left) and PEDOTPSS PCPDTBT:PCBM|Ti (right). P3HT:PCBM and PCPDTBTPCBM (binary blends) create bulk heterojunctions that ensure that all the excitons generated by the incident photons get dissociated into individual electron-hole pairs within their diffusion lengths. Due to a built-in electric field created due to the differences in the work functions of the anode (PEDOTPSS) and cathode (Ti), the newly created electrons and holes are drawn efficiently towards the cathode and the anode side, respectively, minimizing recombination in the active layer (binary blend).

[0032] Figure 6 shows a non-limiting schematic diagram of a fabrication process for an exemplary nanoelectronic device 100. A Si wafer is used as the starting substrate followed by a thin film of Ti / Al (100 nm / 200 nm) deposited thereon using electron-beam evaporation to serve as the sacrificial layer for lifting off the final devices from the substrate. Next, spin coating of organic polymers (PEDOTPSS (-50 nm), as an anode layer, and a binary blend of organic semiconductor (OSC) (-100 nm) of P3HTPCBM or PCPDTBTPCBM forms the OSC layer atop the anode followed by deposition of a thin film of Ti (50 nm) using electron-beam evaporation to form the cathode layer thereon. Next, patterning the micron-sized devices is performed using photolithography and dry-etching techniques, where a 1 pm thick photoresist (AZ3312) is spin-coated on top of Ti (top cathode layer), the patterns made using MLA-150 (mask-less photopatteming) and thereafter the photoresist is allowed to hard-bake before dry etching. For dry-etching, an etchant, such as based on SF6 chemistry, was employed to selectively etch exposed Ti metal. Afterward, oxygen plasma was used to etch away the organic polymers (including the photoresist). For this step, Ti served as a hard-mask for the polymer stack underneath. Finally, releasing the nanoelectronic devices 100 from the Si substrate was achieved by allowing the Al layer underneath to react with diluted TMAH solution (2.7% v / v in water). The released nanoelectronic devices can be collected and stored, for example, in DI water.

[0033] Figure 7A shows a graph of the short circuit current and maximum power generated by nanoelectronic devices (10 pm in diameter) for varied optical intensities. Device structure - PEDOT:PSS|PCPDTBT:PCBM|Ti, laser - 785 nm, values represent median ± SD (n>5).

[0034] Figure 7B shows a power-voltage plot (semi log) for differently sized nanoelectronic devices. The plot shows that the power generated by these devices scale up with an increase in device size. Device structure -PEDOT:PSS|PCPDTBT: PCBM|Ti, intensity7- 6 mW / mm2Legend corresponds to the device diameter. These are representative data from measurements of at least 5 independent devices for each size. Figure 8A shows SEM images of the nanoelectronic devices (i) before release from the substrate and (ii) after release from the substrate. Scale bar - (i) 2 pm; (ii) 4 pm.

[0035] Figure 8B shows a histogram demonstrating the collection yield, providing a quantitative view of the collection efficiency at each step (post TMAH, post APTES, and post NHS-DBCO); values represent mean ± SD (n>5).

[0036] Figure 8C shows a bar graph of the maximum power generated by the nanoelectronic devices before and after release from the substrate using diluted TMAH solution. Device size - 200 pm in diameter, device structure - PEDOT: PSS|P3HT:PCBM|Ti, intensity - 10 mW / mm2(520 nm); values represent median ± SD (n = 5).

[0037] Figure 9 shows a bar graph of the quantitative toxicity analysis of nanoelectronic devices (10 pm in diameter) incubated with wehi-265.1 monocytes. Time varying MTT assay showing biocompatibility of these nanoelectronic devices with minimal toxicity to cells. The x-axis represents the incubation time of the DBCO-functionalized nanoelectronic devices with azide- functionalized monocytes. The concentration of nanoelectronic devices used for the toxicity assessment was varied between 104 to 107 mL-1 and the concentration of cells used was 2 million / mL. For intravenous injection, nanoelectronic devices at a concentration of 106 mL-1 were incubated with cells (2-4 million / mL) for 2 hours. Values represent median ± SD (n=3).

[0038] Figure 10 shows Titanium content as measured in the blood and the brain tissue for quantification of the SWEDs after three-day time-point post self-implantation (as SWEDs contain a 50 nm thin titanium layer) measured by Inductively Coupled Plasma Mass Spectrometry (ICP-MS) when 2 x 106 number of SWEDs only or cell-electronics hybrids were injected into the animal intravenously for the experimental and various control groups. A significant increase in titanium content compared to the baseline was observed only for the brain tissue in the experimental group while the changes in other control groups compared to baseline were insignificant, values represent mean ± SD (n = 3 mice), ***p<0.001 (one-way ANOVA test).

[0039] Figure 11 shows a bar graph of the quantitative toxicity analysis using MTT assay for nanoelectronic devices (10 pm in diameter) incubated with cultured neurons. The plot demonstrates the biocompatibility of these devices with neurons over a period of 24 hours. 50 pl of nanoelectronic devices at various concentrations (105 to 107 mL-1) were incubated with neurons cultured in a 96 well plate for 7-10 days prior to the experiment; values represent mean ± SD (n=3).

[0040] Figures 12A and 12B show fluorescence density histograms of GFAP and Iba-1 levels after nanoelectronic device and PBS injections, respectively. There was an initial increase in GFAP and Iba-1 densities in the vicinity of the injection site within 3 days after injections. GFAP increase reached a plateau 3 to 7 days after injections (no significant difference between 3-day and 7-day cases), while a reduction in Iba-1 density after 7 days was observed. No significant disparities in astrocyte / microglial levels between nanoelectronic devices and PBS injections were discernible, suggesting that the observed immunoreactions of astrocytes and microglia were to the intracranial injection procedure itself but not to the nanoelectronic devices. "‘ns7’ denotes no significant difference (2-way ANOVA test for (c) and student t-test for (d)). Mean ± SD (n=3).

[0041] Figure 13A is a non-limiting schematic diagram of a layout of a biphasic pulse generator circuit having a buried oxide, metal, field oxide, N-active, and poly-silicon components. The entire circuit can be implemented in less than 1 pm2.

[0042] Figure 13B is a non-limiting schematic diagram of a layout of a biphasic pulse generator showing individual layers including silicon oxide, titanium, P3HT:PCBM, platinum, SOI- CMOS, PEDOTTSS, oxide insulator, indium tin oxide (ITO), PCPDTBT:PCBM, and CMOS- Metal.

[0043] Figure 14 shows a non-limiting representation of nanoelectronic device 200. which works on the magnetoelectric principle, and is a core-shell device having a magnetostrictive core 210 and a piezoelectric shell 220.

[0044] Figure 15A is a TEM image showing magnetostrictive cobalt ferrite (CFO) nanotransducers that convert external magnetic field to mechanical energy.

[0045] Figure 15B is a TEM image showing piezoelectric Barium Titanate (BTO) nanotransducers that convert acoustic stimulus into electrical energy.

[0046] Figure 15C is a TEM and corresponding elemental mapping using TEM-EDS for fabricated CFO-BTO core-shell nanostructures. The elemental mapping clearly shows the coreshell morphology of the nanostructures.

[0047] Figure 16 shows magnetization of fabricated magnetostrictive Cobalt Ferrite (CFO) nanotransducers as a function of applied magnetic field measured using a vibrating sample magnetometer (VSM).

[0048] Figure 17 shows measured piezoelectric force microscopy (PFM) displacement and phase loops for piezoelectric BTO nanotransducers.

[0049] Figure 18 shows measured magnetoelectric (ME) coefficient of the CFO-BTO pellet as a function of applied DC bias magnetic field.

[0050] Figure 19 shows functionalization of magnetostrictive, piezoelectric and magnetoelectric devices with immune cells. Scale bar is 20 pm. Figure 20 shows confocal images of mouse brain slice for LPS inflammation model mice with (top) and without (bottom) cell-device hybrid injection through vasculature showing successful self-implantation of devices at the inflammation site. Scale bar is 200 pm.

[0051] Figure 21 shows a schematic diagram of setup for neural stimulation in a freely moving mouse using nanoelectronic devices exposed to magnetic stimulation.

[0052] Figure 22 is a representative image demonstrating that the cell-electronics hybrids reside in the brain parenchyma outside the vasculature (magenta) at the target region. Scale: 100 pm. demonstrating crossing of intact blood-brain-barrier by electronics.

[0053] DETAILED DESCRIPTION

[0054] Nanoelectronic devices and bioelectronic hybrids thereof, as well as methods of making and using thereof are described herein. These devices can be implanted non-surgically in a subject and can be used for various medical applications.

[0055] I. Definitions

[0056] “Magnetostrictive," as used herein, refers to materials which exhibit a property where they can change their shape or dimensions in response to an applied magnetic field. When subjected to a magnetic field, magnetostrictive materials experience mechanical deformation, which can include expansion, contraction, or torsion. This effect is reversible, meaning that when the magnetic field is removed, the material returns to its original shape.

[0057] “Piezoelectric," as used herein, refers to materials which exhibits the piezoelectric effect, which is the ability to generate an electric charge in response to mechanical stress or deformation. When mechanical pressure or stress is applied to a piezoelectric material, it causes a displacement of electric charges within the material, leading to the creation of an electric potential difference or voltage.

[0058] “Modulation,” as used herein, refers to modification or alteration of biological processes and / or functions, within a target site or region of a subject, by the nanoelectronic devices, or hybrids thereof, described, as compared to the biological processes and / or functions prior to any exposure to the nanoelectronic devices.

[0059] “Biocompatible”, as used herein, refers to materials that are, along with any metabolites or degradation products thereof, generally non-toxic to the recipient, and do not cause any significant adverse effects to the recipient, at concentrations resulting from the degradation of the administered materials. Generally speaking, biocompatible materials are materials should not elicit an inappropriate inflammatory or immune response when administered to a patient. “Biodegradable,” as used herein, refers to a material that can be degraded or eroded by enzymatic action or hydrolysis under physiologic conditions into smaller units or chemical species of the original material that are capable of being metabolized, eliminated, or excreted by a subject.

[0060] The term “subject,” as used herein, refers to a mammal, such as a human.

[0061] The terms “treatment” and “treating” refer to the medical management of a subject with the intent to cure, ameliorate, and / or stabilize a disease in the subject. This term includes active treatment toward the improvement of the disease, or palliative treatment designed for the relief of symptoms rather than intended for curing of the disease; preventative treatment is directed to minimizing or partially or completely inhibiting the development of a disease. It is understood that treatment, while intended to cure, ameliorate, and / or stabilize a disease or symptoms thereof, need not actually result in any particular degree of cure, amelioration, stabilization, and / or prevention.

[0062] The term “pharmaceutically acceptable” refers to compositions or other materials and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact wi th the tissues of human beings and animals without excessive toxicity, irntation. allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. The phrase “pharmaceutically acceptable carrier or diluent” refers to pharmaceutically acceptable materials, compositions or vehicles, such as a liquid or solid filler, diluent, solvent or encapsulating material involved in carrying or transporting any subject composition, from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of a subject composition and not injurious to the subject to which it is administered.

[0063] The term “non-invasive or minimally -invasive route of administration” includes intravenous administration, subcutaneous administration, intranasal administration, oral administration, intramuscular administration, or intradermal administration, epidural injection, spinal injection or any other introduction into the bloodstream or the cerebrospinal fluid (CSF) or combinations thereof. Such minimally -invasive administration may include injections through one or more needles known in the medical arts. The “non-invasive or minimally - invasive route of administration” is a non-surgical route of administration.

[0064] Numerical ranges include ranges of thicknesses, ranges of pressures, ranges of molecular weights, ranges of integers, ranges of times, ranges of electric current, ranges of length, ranges of diameters, etc. The ranges disclose individually each possible number that such a range could reasonably encompass, as well as any sub-ranges and combinations of sub-ranges encompassed therein. For example, an activating step may be carried out for a period of time in the range of about 1 min to 10 min. also refers to time values that can be selected independently from about 2, 3, 4, 5, 6, 7, 8, and 9 minutes, as well as any range between these numbers (for example, 3 min to 8 min), and any possible combination of ranges between these time values.

[0065] Use of the term "about" is intended to describe values either above or below the stated value, which the term "‘about’' modifies, to be within a range of approximately + / - 10%. When the term "about" is used before a range of numbers (i.e.. about 1-5) or before a series of numbers (i.e., about 1, 2, 3, 4, etc.) it is intended to modify both ends of the range of numbers and / or each of the numbers recited in the entire series, unless specified otherwise.

[0066] Nanoelectronic devices and hybrid bioelectronic devices are described herein. In some embodiments, the hybrid bioelectronic device includes a wirelessly actuatable component which includes: an external energy harvesting transducer or power source, wherein the external energy harvesting transducer or power source is associated with a biological and / or chemical agent capable of recognizing a target region in a body and self-implanting in the target region, wherein the external energy harvesting transducer or power source, when exposed to an externally applied stimulus, converts at least a portion of the externally applied stimulus into energy.

[0067] Typically, the dimensions of the nanoelectronic device (the wirelessly actuatable component) are in a range of betw een about 5 nanometers to about 5 millimeters. The nanoelectronic device is capable of transport through a fluid system of the body. In some instances, the fluid system is selected, without limitation, from a vascular system (such as blood or plasma), lymph system, central nervous system (such as cerebrospinal fluid), excretory system (such as urine), digestive system (such as bile, stomach acid), salivary system (such as saliva), skeletal system (such as synovial fluid), extracellular matrix and combinations thereof. The nanoelectronic devices are capable of traversing various types of fluids in a body (e.g., a human patent’s body) innocuously and without adverse effects.

[0068] In some instances, the nanoelectronic device includes at least one component which is a transducer which converts the externally applied stimulus into electrical, mechanical, thermal, and / or electro-chemical energy. In some instances, the nanoelectronic device may further include at least one component capable of being powered by the external energy harvesting transducer or power source. The at least one component may provide one or more functionalities to the nanoelectronic device including, but not limited, to sensing, computing, analysis, and / or feedback functionalities. The at least one component can be capable of wirelessly receiving and / or transmitting commands and / or data. The at least one component may be configured and capable of brain-computer interfacing. More than one component may be present on a nanoelectronic device. In still other instances, the at least one component is a nanoelectronic circuit, which comprises an antenna for wireless communication, and is capable of being powered by a transducer which converts the externally applied stimulus into electrical, mechanical, thermal, and / or electro-chemical energy.

[0069] In some instances, the externally applied stimulus is selected from, without limitation, from an optical, electrical, electromagnetic, magnetic, radiofrequency, and / or acoustic stimulus. The parameters of the externally applied stimulus are selected to cause the external energy harvesting transducer to transduce the stimulus into energy. Such energy may be, without limitation, electrical energy, mechanical energy, thermal energy', and / or electro-chemical energy7.

[0070] The nanoelectronic device described herein can be used in a hybrid bioelectronic device (or bioelectronic hybrid) which includes a biological agent and / or chemical agent associated thereto which are capable of recognizing a target region in a body and self-implanting in the target region.

[0071] In some instances, the chemical agent is present as a surface functionalization on at least one surface of the device. In certain instances, at least one surface of the external energy harvesting transducer is functionalized with the chemical agent. In some instances, the nanoelectronic device, or a component thereof (such as the external energy harvesting transducer or at least one component, if present), is associated with (attached to) a biological agent. In some instances, the biological agent is selected from a biological cell, a virus, a cell membrane component, an extracellular vesicle, a synthetic cell, or a biomolecule.

[0072] The nanoelectronic devices alone, and / or as bioelectronic hybrids thereof, can be administered to the subject as part of a pharmaceutical composition. In some non-limiting embodiments, the pharmaceutical composition includes a plurality7of the nanoelectronic devices and / or bioelectronic hybrids thereof dispersed in a pharmaceutically acceptable (liquid) carrier; and optionally one or more pharmaceutically acceptable excipients.

[0073] The nanoelectronic devices and bioelectronic hybrids thereof can be used for various therapeutic, prophylactic, or diagnostic applications. For example, the hybrid bioelectronic device may be used to treat inflammation-related diseases in a subject, as described below. The region of inflammation is the fundamental hallmark and key therapeutic target of many7diseases such as Alzheimer’s disease, multiple sclerosis, ischemic stroke, brain tumor, chronic pain, spinal cord injury7, peripheral nerve injury, paralysis, essential tremor, a movement disorder, cardiovascular diseases, arthritis, or a mental disorder, such as depression / anxiety, schizophrenia, post-traumatic stress disorder, autism, bipolar disorder, and disruptive mood dysregulation disorder. Previous reports have shown that localized electrical modulation of the inflamed region in these diseases can benefit their treatment by addressing fundamental factors contributing to their etiology.

[0074] The nanoelectronic devices and bioelectronic hybrids thereof can each independently be provided as part of a kit. The (sterile) kit is for medical use. The kit may include instructions for preparing a pharmaceutical composition containing the devices and / or hybrids. The kits may include pharmaceutically acceptable carriers / diluents and other excipients for formulating a pharmaceutical composition of the devices or hybrids.

[0075] II. Nanoelectronic Devices and Bioelectronic Hybrids Thereof

[0076] Nanoelectronic devices and bioelectronic hybrids thereof are described in detail below.

[0077] A. Nanoelectronic Devices

[0078] The nanoelectronic device may be a wirelessly actuatable component configured to interact with an externally applied stimulus, wherein the wirelessly actuatable component is in size range from about 5 nanometers to about 5 millimeters. In one non -limiting instance, a nanoelectronic device includes: an external energy harvesting transducer; wherein the external energy harvesting transducer is associated with a biological and / or chemical agent capable of recognizing a target region in a body and self-implanting in the target region; and wherein the external energy harvesting transducer, when exposed to an externally applied stimulus, transduces at least a portion of the externally applied stimulus into energy.

[0079] Non-limiting nanoelectronic devices are shown in Figure 1A, showing nanoelectronic device 100, and Figure 14. showing nanoelectronic device 200. which are each described in detail below. In some instances, the external energy harvesting transducer is fabricated by bottom-up chemical synthesis methods and does not have a substrate. In preferred instances, the external energy harvesting transducer is fabricated by top down microfabrication techniques on a substrate but does not include any substrate on which it was fabricated on, where the external energy harvesting transducer following fabrication is released from the substrate it w as fabricated on and the nanoelectronic device does not include the substrate therein.

[0080] Typically, the dimensions of the nanoelectronic device (the wirelessly actuatable component), such as length, width, diameter are in the range of between about 5 nanometers to about 5 millimeters.

[0081] The nanoelectronic device is capable of transport through a fluid system of the body. In some instances, the fluid system is selected, without limitation, from a vascular system (such as blood or plasma), lymph system, central nervous system (such as cerebrospinal fluid), excretory system (such as urine), digestive system (such as bile, stomach acid), salivary system (such as saliva), skeletal system (such as synovial fluid), extracellular matrix and combinations thereof. The nanoelectronic devices are capable of traversing various types of fluids in a body innocuously and without adverse effects.

[0082] In some instances, the nanoelectronic device includes at least one component which is a transducer which converts the externally applied stimulus into electrical, mechanical, thermal, and / or electro-chemical energy. In some instances, the nanoelectronic device includes at least one component capable of being powered by the external energy harvesting transducer or power source. This component may provide one or more functionalities to the hybrid bioelectronic device including, but not limited, to modulation of biological functions, sensing, computing, analysis, and / or feedback functionalities. This component can be capable of wirelessly receiving and / or transmitting commands and / or data. This component may be configured and capable of brain-computer interfacing. More than one component may be present on a nanoelectronic device. In still other instances, the at least one component is a nanoelectronic circuit, which comprises an antenna for wireless communication, and is capable of being powered by a transducer which converts the externally applied stimulus into electrical, mechanical, thermal, and / or electro-chemical energy . Methods of fabricating and including such component(s) into the nanoelectronic device include processes, such as bottom-up chemical synthesis or top-down microfabrication techniques.

[0083] In some instances, the externally applied stimulus is selected from, without limitation, from an optical, electrical, electromagnetic, magnetic, radiofrequency, and / or acoustic stimulus. The parameters of the externally applied stimulus are selected to cause the external energy harvesting transducer to transduce the stimulus into energy. Such energy may be, without limitation, electrical energy, mechanical energy, thermal energy', and / or electro-chemical energy7.

[0084] In some instances, the energy transduced / generated is electrical energy, and then the transducer acts as an external energy harvesting power source.

[0085] In some instances, the nanoelectronic device is a planar device with a total thickness in the range of 1 nanometer to 500 micrometers and lateral size in the range of 5 nanometers to 5 millimeters.

[0086] In some instances, the nanoelectronic device may include at least one surface functionalization present on at least one surface of the nanoelectronic device and / or surface of the external energy7harvesting transducer. In some instances, at least one surface of the nanoelectronic device is functionalized with chemical functionalities which are reactive with a complementary7group to allow attachment of the surface to another material. For example, azide groups may be functionalized on surface(s) of the device (or components thereof) which can react with dibenzocyclooctyne via a click reaction. Various ty pes of methods for chemically functionalizing surface(s) of devices (or components thereof) are known in the art.

[0087] The nanoelectronic devices are considered biocompatible. In some instances, they may be biodegradable which allows for their degradation into physiologically and environmentally benign by-products after the required application period is over, to avoid the need for explantation or extraction of the nanoelectronic devices, or any components or bioelectronic devices thereof.

[0088] 1. Optical Transducers

[0089] 1.1 Organic Semiconductor-Based Optical Transducers

[0090] In some non-limiting instances, the external energy harvesting transducer is an organic semiconductor based photovoltaic transducer which encompasses various device structures, including single layer to multiple-layer configurations. A single layer can include organic semiconductor only. Additional layers may include apart from the organic semiconductor, anode, cathode, electron transport layers (ETLs), hole transport layers (HTLs), additional electrode layers for improving interface properties as well as protective layers. In preferred embodiments, the photovoltaic transducer does not include any substrate on which it was fabricated, where the photovoltaic transducer following fabrication is released from the substrate it was fabricated on and the nanoelectronic device does not include the substrate therein.

[0091] Figure 1A shows a nanoelectronic device operating on a photovoltaic principle which is a three-layered device based on organic polymers where layer 110 is an anode, layer 120 is an organic semiconductor, and layer 130 is cathode.

[0092] 1.1.1 Multi-Layer Device Configurations

[0093] In preferred embodiments, multi-layer devices are implemented to optimize charge extraction and overall device performance. Example configurations include: (a) a Basic Three- Layer Structure: Anode, Active layer (organic semiconductor blend). Cathode, or (b) a Five- Layer Structure: Anode, Hole transport layer (HTL), Active layer (organic semiconductor blend); Electron transport layer (ETL), Cathode.

[0094] 1.1.2 Example Device Configurations

[0095] The following non-limiting examples illustrate specific embodiments of the multi-layer device:

[0096] • Example 1:

[0097] PEDOT:PSS (Anode) I PM6:L8-BO (Active Layer, comprising a semiconducting polymer blend) / PNDIT-F3N (ETL) / Ti (Cathode) / IrOx (Neural interfacing electrode) • Example 2:

[0098] PEDOTPSS (Anode) / PSBTBTPC70BM (Active Layer, comprising a semiconducting polymer blend) / Ti (Cathode) / TiN (Neural interfacing electrode)

[0099] • Example 3:

[0100] PEDOTPSS (Anode) / PCDTBTPC70BM (Active Layer, comprising a semiconducting polymer blend) / Ti (Cathode) / TiN (Neural interfacing electrode)

[0101] • Example 4 (Inverted Architecture):

[0102] ITO (Cathode) / ZnO (ETL) / PSBTBTPC70BM (Active Layer, comprising a semiconducting polymer blend) / MoOs (HTL) I Ag (Anode)

[0103] • Example 5 (Inverted Architecture):

[0104] ITO (Cathode) / ZnO (ETL) / Active Layer / PEDOTPSS (HTL) / Ag (Anode)

[0105] In various embodiments, the layer thicknesses and material compositions can be adjusted to yield a device optimized for performance and efficiency.

[0106] 1.1.3 Materials

[0107] 1.1.3.1 Organic Semiconductor Polymers for Active Layer

[0108] The active layer includes an acceptor material and a donor material. Typically, the active layer is formed of a blend of an acceptor material and a donor material. Such donor and acceptor materials include, without limitation: a) Donor Materials: poly[(2,6-(4,8-bis(5-(2-ethylhexyl-3-fluoro)thiophen-2-yl)-benzo[l,2-b:4,5-b']dithiophene))-alt- (5, 5-(l',3'-di -2 -thienyl-5',7'-bis(2-ethylhexyl)benzo[l',2'-c:4',5'-c']di thiophene-4, 8-dione)] (PM6). poly[(2,6-(4.8-bis(5-(2-ethylhexyl-3-fluoro)thiophen-2-yl)-benzo[l,2-b:4,5- b']dithiophene))-alt-(5,5-(r,3'-di-2-thienyl-5',7'-bis(2-ethylhexyl)benzo[r,2'-c:4',5'- c'] dithiophene-4, 8-dione)] (DI 8), poly[(4,4'-bis(2-ethylhexyl)dithieno[3,2-b:2',3'-d]silole)-2,6- diyl-alt-(2,l,3-benzothiadiazole)-4,7-diyl] (PSBTBT), poly[N-9'-heptadecanyl-2.7-carbazole-alt- 5,5'-(4',7'-di-2-thienyl-2',l',3'-benzothiadiazole)] (PCDTBT), poly(3-hexylthiophene) (P3HT), poly(2,6-(4,4-bis-(2-ethylhexyl)-4H-cyclopenta[2,l-b:3,4-b’]dithiophene)-alt-4,7-(2,l,3- benzothiadiazole) (PCPDTBT), poly(2,5-thiophene) (P3T), poly(3,4-ethylenedioxythiophene) (PEDOT), poly(9,9-dioctylfluorene) (PFO), poly(2,5-bis(3-alkylthiophen-2-yl)thieno[3,2- b]thiophene) (PBTTT), poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOTPSS), poly(3.6-dioctylthieno[3,2-b]thiophene) (P3OT), poly(2,5-thiophene vinylene) (PTV). polythiophenes with various alkyl side chains and substitutions, poly(3,4-difluorothiophene-co- bithiophene) (PFTB), poly(2,6-carbazole) (PCz), poly(9,9-dioctylfluorene-co-bithiophene) (F8T2), and combinations thereof. In some instances, the donor material is selected from poly

[0109] (3 -hexylthiophene) (P3HT) or poly[2,6-(4,4-bis-(2-ethylhexyl)-4H-cyclopenta[2.1-b;3,4- b']dithiophene)-alt-4,7(2,l,3-benzothiadiazole)] (PCPDTBT) b) Acceptor Materials:

[0110] L8-BO (5,5'-Bis((2-butyloctyl)oxy)-[l ,T:3',l"-terphenyl]-4,4"-dicarbaldehyde), Y6 (2,2'- ((2Z,2'Z)-((12, 13-bis(2-ethylhexyl)-3,9-diundecyl-l 2, 13-dihydro-[l ,2,5]thiadiazolo[3,4- e]thieno[2,3":4',5']thieno[2',3':4,5]pyrrolo[3,2-g]thieno[2',3':4,5]thieno[3.2-b]indole-2,10- diyl)bis(methanylylidene))bis(5.6-difluoro-3-oxo-2.3-dihydro-lH-indene-2,l- diylidene))dimalononitrile), Z8 (2,2'-((2Z,2'Z)-((l 2, 13-bis(2-ethylhexyl)-3,9-diundecyl-l 2, 13- dihydro-[l,2,5]thiadiazolo[3,4-e]thieno[2,3":4',5']thieno[2',3':4,5]pyrrolo[3,2- g]thieno[2',3':4,5]thieno[3,2-b]indole-2,10-diyl)bis(methanylylidene))bis(3-oxo-2,3-dihydro-lH- indene-5.6-dicarbonitrile))dimalononitrile), o-BTP-eC9 (2,2'-((2Z,2'Z)-((12,13-bis(2- ethylhexyl)-3,9-bis(2-(2-ethoxyethoxy)ethyl)-12,13-dihydro-[l,2,5]thiadiazolo[3,4- e]thieno[2,3":4',5']thieno[2',3':4,5]pyrrolo[3,2-g]thieno[2',3':4,5]thieno[3,2-b]indole-2,10- diyl)bis(methanylylidene))bis(3-oxo-2,3-dihydro-lH-indene-2,l-diylidene))dimalononitrile), P- 2BTh-F (3.9-bis(2-methylene((3-(l,l-dicyanomethylene)-6,7-difluoro)indanone))-5,5,l 1,11- tetrakis(4-hexylphenyl)dithieno[2.3-d:2',3'-d']sindaceno[l,2-b:5,6-b']dithiophene), and PC70BM ([6,6]-Phenyl-C71 -butyric acid methyl ester, PCBM (Phenyl-C61 -butyric acid methyl ester), ITIC (Indacenodithieno[3,2-b]thiophene-2,8-dicarboximide), IT-M (Indacenodithiophene-based molecule), EH-IDTBR (2,2’-((2Z,2’Z)-((12,13-bis(2-ethylhexyl)-3,9-diundecyl-12,13-dihydro- [l,2,5]thiadiazolo[3,4-e]pyrazine-2,2,6,6-tetrayl))bis(4,5-dihydrothieno[3,4-b][1.4]dioxine-5,7- diyl))bis(methanylylidene))bis(3-ethyl-2-thioxothiazohdin-4-one), and combinations thereof. In some instances, the acceptor material is [6,6]-phenyl-C61-butyric acid methyl ester (PCBM).

[0111] In some instances, the thickness of the active layer may be in a range of about 10 nm to about 200 nm, as well as sub-ranges or individual values contained within. The relative amounts of acceptor to donor materials in an active layer may be varied. In some instances, the amount of acceptor and donor materials are in a range of about 10 to 90% by weight of the active layer, where the sum of the percentages of both materials totals 100%. The donor and acceptor materials may be chosen based on their absorption spectra and absorption properties (such as absorption coefficients).

[0112] 1. 1.3.2 Electrode Materials for Anode and Cathode

[0113] The anode and the cathode can be formed of any suitable conductive material. Typically, the anode and the cathode may be formed from the same or different conductive materials. In some instances, the conductive material can be made of or contain a conductive organic polymer, such as of poly(3,4-ethylenedioxythiophene) polysty rene sulfonate (PEDOT:PSS), poly(3.4-ethylenedioxythiophene), polyacetylene. poly(3-alkyl-thiophene). polyaniline, polyisothianaphthalene, poly-(p-phenylene), poly-(p-phenylene vinylene), polypyrole, polythiophenes, poly(3,4-ethylenedioxythiophene) tosylate , poly(9,9-dioctylfluorene), poly(3,4- dioctyloxythiophene), poly(3-hexylthiophene), or combinations thereof. In some instances, cathode material is selected from, without limitation, titanium, graphene, aluminum, calcium, barium, lithium fluoride, transition metals (such as platinum), platinum-iridium alloys, low work function metals and alloys thereof, organic materials, such as BCP (bathocuproine) and BPhen (4,7-diphenyl-l,10-phenanthroline), evaporated organic films, like C60 and C70, or combinations thereof. In some other instances, the anode and cathode may be formed from a conductive material which is a composite ink containing graphene, carbon black, or carbon nanotubes. Such graphene composite inks are commercially known and can be formulated with a polymer, such as a polyester (such as PLGA). Other suitable conductive materials are known and can be commercially purchased. In some instances, the cathode can be formed of or includes titanium or graphene. In some instances, the anode material can be formed of or includes poly(3.4-ethylenedioxythiophene): polystyrene sulfonate (PEDOT:PSS).

[0114] In some instances, the anode and cathode can be spin-coated, deposited, or 3-D printed from a suitable conductive material onto a substrate.

[0115] In some instances, the anode and cathode electrodes can independently have any suitable thickness ranging from about 1 to 500 nm, as well as sub-ranges or individual values contained within. The thickness of the electrodes can be uniform or non-uniform.

[0116] The anode and cathode materials may be chosen based on their work functions and differences thereof, which create a built-in electric field in the device that draws electrons and holes formed in the active layer towards the anode and cathode sides and minimizes recombination events.

[0117] 1. 1.3.3 Electron Transport Layer (ETL) Materials

[0118] In various embodiments, the electron transport layer (ETL) of the photovoltaic transducer is configured to facilitate the efficient extraction and transport of electrons from the active layer. The ETL may include one or more suitable materials for promoting electron mobility and reducing recombination. In some instances, the ETL includes one or more of the following materials: Poly[[2,7-bis(2-ethylhexyl) l,2,3,6.7.8-hexahydro-l,3,6,8-tetraoxobenzo[lmn]phenanthroline-4,9-diyl]-

[0119] 2.5-thiophenediyl[9,9-bis[3'-((N,N-dimethyl)-N-ethylamino)propyl]-9H-fluorene-2,7-diyl]-

[0120] 2.5-thiophenediyl] (PNDIT-F3N), Zinc oxide (ZnO), Titanium dioxide (TiCh) Depositing the ETL includes methods such as spin-coating, vapor deposition, etc. In some embodiments, the ETL is formed with a thickness ranging from about 1 to 100 nm, with the specific thickness chosen to optimize electron extraction and overall device performance.

[0121] 1. 1.3.4 Hole Transport Layer (HTL) Materials

[0122] The hole transport layer (HTL) is configured to promote efficient hole extraction from the active layer to the anode while minimizing recombination losses. In various embodiments, the HTL may comprise any suitable material to support hole transport. Suitable HTL materials include, but are not limited to: Poly(3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS), Molybdenum trioxide (MoCh), Tungsten trioxide (WOs), Vanadium pentoxide (V2O5).

[0123] The HTL materials can be deposited by methods such as spin-coating, vapor deposition, or printing. The layer thickness and deposition conditions may be adjusted to achieve the desired electrical characteristics and device efficiency.

[0124] 1. 1.3.5 Neural Interfacing Electrode Materials

[0125] For embodiments incorporating neural interfacing, the photovoltaic device may include one or more neural interfacing electrodes. These electrodes are selected based on their biocompatibility, electrical conductivity, and ability to form stable interfaces with biological tissue. In various embodiments, the neural interfacing electrodes may include materials selected from the following groups: a) Metals and Metal Oxides -Titanium Nitride (TiN), Platinum (Pt), Iridium Oxide (IrOx), including Sputtered Iridium Oxide Film (SIROF) and Activated Iridium Oxide Film (AIROF), Ruthenium Oxide (RuOx), Platinum-Iridium Alloy (Pt-Ir), Tantalum / Ta2O5, Gold (Au) and porous Au, Oxidized forms such as AuOx, PtOx, or Palladium Oxide (PdOx), Tungsten (W), Nano Pt b) PEDOT:PSS and Variants - PEDOT:PSS, PEDOT:PSS / Ptlr, PEDOT / CNT (incorporating Carbon Nanotubes) c)Carbon-Based Materials - Glassy Carbon, Porous Graphene, Carbon Nanotube (CNT) Fibers, Graphene Fibers, Reduced Graphene Oxide, Hybrid Diamond / Carbon Fiber, 3D Fuzzy' Graphene.

[0126] The specific material or combination of materials may be selected based on the desired balance of electrical performance, mechanical stability, and biocompatibility for neural interfacing applications.

[0127] 1.2 Inorganic Optical Transducers

[0128] In some non-limiting instances, the external energy harvesting or signal transducing device is configured as an inorganic optical transducer. In certain embodiments, the inorganic optical transducer may be fabricated at subcellular scales. In preferred embodiments, the inorganic optical transducer does not include any permanent substrate upon which it was originally fabricated: rather, it is released from such a substrate such that the resulting nanoelectronic device may be free-standing or integrated into biological tissues without including the substrate therein. In some instances, these inorganic optical transducers can be encapsulated with biocompatible coatings (e.g., parylene) to facilitate stable interfaces with living cells and tissues.

[0129] The inorganic optical transducers described herein can operate by converting incident light (including visible or near-infrared wavelengths) into electrical signals. These devices can operate via non-Faradaic to Faradaic mechanisms of action and demonstrate high efficiency in charge separation and transfer. The thickness of any layers within these devices can be uniform or non-uniform and, in some instances, can be selected to optimize device performance.

[0130] 1.2.1 Inorganic Semiconductor-Based Optoelectronic Devices

[0131] In some instances, the inorganic optical transducer includes:

[0132] Silicon Nanowires (NWs) with p-i-n Junctions: In certain embodiments, silicon nanowires are fabricated with doping profiles to create p-i-n junctions. Methods of forming these nanowires are not particularly restricted and may include bottom-up synthesis, such as vapor-liquid-solid (VLS) growth.

[0133] III-V Semiconductor Nanowires (e g., GaAs, InP): In some instances, III-V nanowires provide enhanced absorption due to direct bandgaps.

[0134] Heterostructure Nanowires Combining Different Semiconductor Materials: In certain embodiments, heterostructure nano wires incorporate metal-semiconductor junctions (for example, Au-TiCh NW). Such configurations may improve charge separation, reduce recombination, or enhance device stability.

[0135] Thin Film Planar Devices: In some instances, thin film planar devices, such as singlejunction GaAs thin-film solar cells, amorphous or crystalline silicon solar cells, or other thin film solar cell technologies (e.g., Copper Indium Gallium Selenide (CIGS) and Cadmium Telluride (CdTe)).

[0136] 1.2.2 Two-Dimensional Material-Based Optoelectronic Devices

[0137] In some non-limiting embodiments, two-dimensional (2D) materials are used to form the inorganic optical transducer. Such 2D materials may include, but are not limited to, graphene, transition metal dichalcogenides (TMDs), or layered heterostructures combining multiple 2D materials. The thickness of the 2D material stack can be varied (e.g., from a single monolayer to multiple layers) to tailor the optical and electronic properties. In preferred embodiments, the device structure can be tuned to optimize light absorption and charge transport. 1.2.3 Quantum Dot-Based Optoelectronic Devices

[0138] In certain embodiments, the inorganic optical transducer includes quantum dot-based optoelectronic devices. Quantum dots may be formed of:

[0139] Colloidal Quantum Dots: Non-limiting examples include PbS, CdSe, or other semiconductor materials with tunable bandgaps.

[0140] Hybrid Quantum Dot-2D Material Systems: In some instances, quantum dots (e.g., CdTe) are combined with 2D materials to leverage strong excitonic effects, broad absorption ranges, or enhanced charge mobility.

[0141] 1.2.4 Plasmonic Nanostructure-Enhanced and Perovskite-Based Nanostructured Devices

[0142] In other non-limiting instances, the inorganic optical transducer may include either: Plasmonic Nanostructure-Enhanced Devices: Incorporating plasmonic nanostructures that enhance local electromagnetic fields.

[0143] Perovskite-Based Nanostructured Devices: Formed of metal halide perovskites or related materials, which offer high absorption coefficients, efficient charge transport, and can be processed in a range of thicknesses.

[0144] 1.2.5 Porphyrin-Based and Semiconducting Polymer Nanoparticle Devices

[0145] In yet other non-limiting instances, the inorganic optical transducer may further include: Porphyrin-Based Optoelectrodes: Non-limiting examples include zinc porphyrin nanorods coated with TiO2 (ZST) and bimetallic zinc and gold porphyrin nanosheets (ZnAuPN). Porphyrins may be selected for their strong light absorption and potential for efficient charge transfer.

[0146] Semiconducting Polymer Nanoparticles: In some embodiments, semiconducting polymer nanoparticles, such as P3HT nanoparticles, or junction-based nanomaterials (e.g., aluminum antimonide (AlSb) nanocrystals), are included. These materials can be optimized for biocompatibility, absorption spectra, and processability.

[0147] 1.2.6 Fabrication and Integration

[0148] In preferred embodiments, any of the foregoing inorganic optical transducers may be fabricated using bottom-up synthesis (e.g., vapor-liquid-solid growth for nanowires), top-down fabrication (e.g., lithography and etching), solution-based processing (e.g.. for quantum dots, 2D materials, or polymer nanoparticles), or hybrid approaches combining multiple techniques. These transducers can be encapsulated with inert or biomedical coatings (e.g., pary lene) to ensure biocompatibility'. The device size such as length, width, diameter are typically in the range of between about 5 nanometers to about 5 millimeters and for planar devices the thickness is in the range of 1 nm to 500 pm. 2. Magnetostrictive, Piezoelectric and Magnetoelectric Transducer

[0149] In some non-limiting instances, the external energy transducer comprises magnetostrictive (MS) or piezoelectric (PE), or magnetoelectric (ME) transducers that convert electromagnetic, magnetic and acoustic fields into mechanical, thermal, or electrical energy.

[0150] Figure 14 illustrates a nanoelectronic device operating on a magnetoelectric principle with a core-shell structure where a magnetostrictive core 210 is surrounded by a piezoelectric shell 220, enabling conversion of magnetic fields into electric fields using strain coupled interfaces.

[0151] These transducers can be fabricated through both bottom-up and top-down approaches. Bottom-up approaches may include chemical synthesis methods such as coprecipitation and solgel processing, while top-down approaches may employ conventional microfabrication techniques followed by substrate release.

[0152] The size, shape, and configuration of these transducers are not particularly restricted and can range from 5 nm to 5 mm in lateral dimensions. For planar devices the thickness ranges from 1 nm to 500 pm. The relative proportions of different materials in multi-component transducers, such as the core-shell ratio in magnetoelectric devices, can also be varied as needed.

[0153] In some instances, the transducers may include top and bottom electrodes formed from any suitable conductive material. The transducers can also be encapsulated in one or more materials that prevent or reduce exposure to wet environments. a. Magnetostrictive Transducer

[0154] Magnetostrictive transducers convert magnetic energy into mechanical energy through magnetostriction. The magnetostrictive material it is formed from is not particularly restricted. In some instances, the magnetostrictive transducer can be formed through chemical synthesis, such as coprecipitation of cobalt ferrite (CoFe2O4, CFO) from metal nitrate precursors. For example, the synthesis may involve combining iron nitrate and cobalt nitrate solutions under controlled conditions, followed by precipitation using a base solution. The synthesis conditions, including but not limited to temperature, pH, concentration of precursors, and reaction time, can be adjusted to control the size and properties of the magnetostrictive transducer. b. Piezoelectric Transducer

[0155] Piezoelectric transducers convert mechanical energy from acoustic fields into electrical energy through the piezoelectric effect. The piezoelectric material it is formed from is not particularly restricted. In some instances, the piezoelectric transducer can be formed through chemical synthesis, such as sol-gel processing of barium titanate (BaTiO3, BTO). The sol-gel synthesis may involve hydrolysis and condensation of metal alkoxide precursors under controlled conditions. Process parameters including but not limited to precursor type, concentration, pH, temperature, and aging time can be varied to achieve desired properties of the piezoelectric transducer. c. Magnetoelectric Transducer

[0156] The magnetoelectric transducer can be formed either as a composite of magnetostrictive and piezoelectric materials or as a single-phase material exhibiting intrinsic coupling between magnetic and electric properties.

[0157] In composite structures, the magnetoelectric transducer combines both magnetostrictive and piezoelectric materials to enable a two-stage energy conversion process. The magnetostrictive material first converts magnetic energy to mechanical strain, which is then transformed into electric fields by the piezoelectric material through strain-coupled interfaces. In some instances, the magnetoelectric transducer can comprise a cobalt ferrite core and barium titanate shell heterostructure. The core-shell structure may be fabricated through sequential synthesis steps, where the magnetostrictive core is first synthesized through coprecipitation, followed by the formation of the piezoelectric shell through sol-gel processing. The synthesis conditions can be optimized to control the interface between the core and shell materials and their relative proportions.

[0158] In single-phase structures, the magnetoelectric transducer comprises materials that exhibit intrinsic coupling between magnetization and electric polarization. In some instances, the magnetoelectric transducer can be formed from materials such as bismuth ferrite (BiFeO3, BFO) that demonstrate inherent magnetoelectric properties without the need for composite structures.

[0159] B. Bioelectronic Hybrids

[0160] A hybrid bioelectronic device (bioelectronic hybrids) includes one or more of the nanoelectronic device (wirelessly actuatable components) described above and a biological or chemical component (or agent) attached thereto, wherein the biological or chemical component is capable of recognizing a target region in a body and self-implanting in the target region, such that the hybrid bioelectronic device is configured to be administered into a body of a subject and thereafter recognize, via the biological or chemical component, a target region within the body and self-implant at the target region.

[0161] In some instances, the chemical agent is present as a surface functionalization on at least one surface of the device. In certain instances, at least one surface of the external energy harvesting transducer is functionalized with the chemical agent. In some instances, the nanoelectronic device, or a component thereof (such as the external energy harvesting transducer or at least one component, if present), is associated with (attached to) a biological agent. In some instances, the biological agent is selected from a biological cell, a virus, a cell membrane component, an extracellular vesicle, a synthetic cell, or a biomolecule. In some instances, the biological agent is genetically engineered and / or biologically or chemically modified.

[0162] In some instances, the nanoelectronic device, or a component thereof, is covalently associated to the biological cell. Figure 1C shows an SEM image of a nanoelectronic device attached to a monocyte forming an example of a bioelectronic hybrid.

[0163] The biological cell typically has a maximum diameter in the range of between 5 to 100 microns. In some instances, the biological cell is a cell selected from immune cells (such as monocytes, macrophages, T cells, neutrophils, and Natural Killer cells), stem / progenitor cells (such as MSCs, iPSCs, HSCs, and neural progenitor cells), glial cells (such as Schwann cells), structural cells (such as epithelial cells), or blood cells (such as red blood cells), or combinations thereof. In some other instances, the biological cell is a genetically engineered cell and / or biologically or chemically modified. The skilled person is familiar with techniques for genetically engineering and / or chemically / biologically modifying cells. In certain instances, the biological cells can be collected from a subject / patient themselves, manufactured from stem cells, or obtained from donors or from umbilical cord blood. In certain instances, the nanoelectronic device is equal to or smaller than the size of a biological cell and is associated to a single biological cell.

[0164] Typically, the biological agent and / or chemical agent provides targeting functionality for transporting the nanoelectronic device to a target site or region in a subject. For example, a biological cell may target inflammation and may allow the bioelectronic hybrid to transport to and target sites of inflammation. In certain instances, the bioelectronic hybrid self-implants onto a target site or region in a body due to the presence of the biological cell.

[0165] In certain instances, the nanoelectronic device, or components thereof, is associated to the biological and / or chemical agent via attachment linkers, and the attachment linkers are either permanent or optionally self-degrading or cleavable by exposure to an external stimulus, such as a biological stimulus. Various suitable attachment linkers are known in the art. In some instances, the external energy harvesting transducer is associated to the biological and / or chemical agent via attachment linkers, and the attachment linkers are optionally self-degrading or cleavable by exposure to an external stimulus, such as a biological stimulus. In still other instances, the external energy harvesting transducer is covalently associated to the biological and / or chemical agent.

[0166] In some instances, the nanoelectronic device, or components thereof, can further include one or more attachment linkers for linking to a target site or region in a body and the attachment linkers may be self-degrading or cleavable by exposure to an external stimulus, such as biological stimulus. In certain instances, after the targeting is achieved, based on specific application needs, the nanoelectronic devices can be designed to either continue to remain attached to the biological cell or can be released through the use of cleavable linkers which either self-degrade after a particular period or are cleavable with external fields or specific biological stimuli (such as pH, proteases, other biomolecules) occur.

[0167] In some instances, the biological agent of the bioelectronic hybrid enables the bioelectronic hybrid to transport across different barriers such as the blood brain barrier (BBB), (cerebrospinal fluid) CSF-brain barrier, CSF-DRG (dorsal root ganglion) barrier and CSF-spinal cord barrier.

[0168] C. Pharmaceutical Compositions Containing the Nanoelectronic Devices or Bioelectronic Hybrids Thereof

[0169] The nanoelectronic devices alone, and / or as bioelectronic hybrids thereof, can be administered to the subject as part of a pharmaceutical composition. In some non-limiting instances, the pharmaceutical composition includes: a plurality of the nanoelectronic devices and / or bioelectronic hybrids thereof: optionally a pharmaceutically acceptable carrier; and optionally one or more pharmaceutically acceptable excipients.

[0170] In some instances, the plurality comprises at least two different types of nanoelectronic devices, which can independently transduce at least two different types of externally applied stimulus into energy. Such energy may be, without limitation, electrical energy, mechanical energy, thermal energy, and / or electro-chemical energy. In other words, the pharmaceutical composition may include at least two types of devices which transduce different wavelengths of optical stimulus into energy7, or which transduce different types of stimulus, such as optical and magnetic stimuli, together in one composition. This allows for a variety of nanoelectronic devices and / or bioelectronic hybrids thereof to be administered to a subject where each different ty pe of nanoelectronic device and / or bioelectronic hybrid thereof is individually addressable by a particular externally applied stimulus. This allows for multiplexing of the different nanoelectronic devices and / or bioelectronic hybrids thereof administered to the subject, as appropriate, by selectively applying the appropriate externally applied stimulus, or combination of stimuli, to generate electrical energy of any particular type of device or hybrid administered. In general, pharmaceutical compositions include effective amounts of the nanoelectronic devices and / or as bioelectronic hybrids thereof. The pharmaceutical compositions may optionally include pharmaceutically acceptable diluents or carriers, such as phosphate buffered saline (PBS), Ringer’s solution, 5% dextrose in water (D5W), and normal / physiological saline (such as 0.9% NaCl). Electrolytes such as, but not limited to, sodium chloride and potassium chloride may also be included in the therapeutic composition. A wide variety of suitable formulations of pharmaceutical composition are known (see, e.g.. Remington’s Pharmaceutical Sciences, 22nded. 2012)). Pharmaceutically acceptable excipients may be included in the pharmaceutical composition(s), which can be selected without limitation: buffers (such as citrate, phosphate, carbonate buffers), preservatives, vitamins, surfactants, liposomal materials (e.g., DSPC), proteins (e.g. albumin, lactose), alcohols, sugars, anti-oxidants (e.g., butylated hydroxyanisole), anti-inflammatories, salts, cholesterol, or amino acids. Other excipients, such inactive or active agents known for pharmaceutical use may also be present. For instance, active agents, such as therapeutic, prophylactic, and / or diagnostic agents known in the art may be included in the composition.

[0171] The pharmaceutical composition containing the nanoelectronic devices alone, and / or as bioelectronic hybrids thereof, can be administered to a subject by any suitable means. In most instances, the route of administration is a nearly non-invasive or minimally invasive procedure such as intravenous, through CSF, intramuscular, subcutaneous, intranasal, oral, epidural, spinal or intradermal routes. Those skilled in the art are familiar with methods of preparing pharmaceutical compositions for different routes of administration.

[0172] In some instances, the pharmaceutical composition includes about 1 million to 100 million nanoelectronic devices and / or bioelectronic hybrids thereof per milliliter of pharmaceutical composition, as well as sub-ranges or individual values contained therein. The nanoelectronic devices and / or bioelectronic hybrids thereof present may be suspended in the carriers / diluents of the composition.

[0173] In some forms, it may be advantageous to formulate pharmaceutical compositions in dosage unit form for ease of administration and uniformity of dosage. ‘"Dosage unit form” as used herein refers to physically discrete units suited as unitary dosages for the subjects to be treated; each unit containing a predetermined quantity of nanoelectronic devices and / or bioelectronic hybrids in order to produce an effect in the subject. The pharmaceutical composition may be administered on any suitable or routine schedule. a. Optional Therapeutic, Prophylactic, or Diagnostic Agents

[0174] In some forms, the pharmaceutical compositions can optionally include one or more additional active agents. Therefore, in some forms, the pharmaceutical composition includes nanoelectronic devices and / or bioelectronic hybrids, in addition to at least one of a therapeutic, prophylactic, and / or diagnostic agents, which agents are known in the art. The additional agents can be included together with the nanoelectronic devices and / or bioelectronic hybrids or may be a separate composition for co-administration. In certain instances, therapeutic, prophylactic, and / or diagnostic agents may be associated to the nanoelectronic devices and / or bioelectronic hybrids themselves, such as by way of a coating present on the devices or hybrids or may be releasably attached thereto. b. Oral Compositions

[0175] The pharmaceutical composition containing the nanoelectronic devices and / or bioelectronic hybrids thereof may be provided in a form suitable for oral administration to a subject in need thereof. In some instances, the pharmaceutical composition contains only nanoelectronic devices without biological cells associated thereof. Oral administration may involve swallowing, so that the nanoelectronic devices and / or bioelectronic hybrids thereof and optionally additional active agent(s) enter the gastrointestinal tract, or buccal or sublingual administration may be employed by which the nanoelectronic devices and / or bioelectronic hybrids thereof enter the blood stream directly from the mouth. Compositions suitable for oral administration include solid compositions such as tablets, capsules containing particulates, liquids, and liquid compositions. Solid formulations containing the nanoelectronic devices and / or bioelectronic hybrids thereof for oral administration may be formulated to be immediate and / or modified release. Modified release formulations include delayed, sustained, pulsed, controlled, targeted and programmed release formulations. c. Parenteral Compositions

[0176] The pharmaceutical composition containing the nanoelectronic devices and / or bioelectronic hybrids thereof may be provided in a form suitable for parenteral administration to a subject in need thereof, through any suitable parenteral route known in the art, including but not limited to intravenous, intramuscular, subcutaneous, intradermal, intranasal, cerebrospinal fluid (CSF), epidural, spinal routes or combinations thereof. Such compositions may include one or more suitable pharmaceutically acceptable vehicles know n in the art for use in administering substances to subjects by any suitable parenteral route known in the art, including but not limited to phosphate buffered saline, optionally with one or more suitable excipients. III. Methods of Making and Characterizing Nanoelectronic Devices and Bioelectronic Hybrids Thereof

[0177] The nanoelectronic devices and / or bioelectronic hybrids thereof described above can be prepared and characterized as described below. a. Preparation of Nanoelectronic Devices

[0178] In some embodiments, the nanoelectronic device or external energy harvesting transducer described above may instead be fabricated using:

[0179] 1. Bottom-Up Synthesis (e.g., vapor-liquid-solid growth of nanowires, nanoparticle synthesis through coprecipitation or sol-gel processing),

[0180] 2. Top-Down Fabrication (e.g.. lithography and etching from bulk wafers),

[0181] 3. Solution Based Processing (e.g.. spin coating of quantum dots, 2D materials, or polymer nanoparticles), or

[0182] 4. Hybrid Approaches combining multiple techniques.

[0183] These methods allow for tailoring of the device's properties, geometry7, material composition, and overall dimensions to optimize performance and biocompatibility7. In some embodiments, the nanoelectronic devices can be encapsulated or functionalized with biocompatible coatings (for example, pary lene), thereby enhancing stability and ensuring minimal adverse interaction with biological systems.

[0184] In some non-limiting instances, a method of preparing substrate free nanoelectronic device comprises the steps of:

[0185] Depositing a Sacrificial Layer on a suitable substrate (for example, a silicon wafer). In some instances, the sacrificial layer comprises aluminum or another material that can be removed by an etchant (e.g., tetramethylammonium hydroxide (TMAH) solution, XeF2, or other chemical etchants).

[0186] Depositing Sequential Device Layers onto the sacrificial layer. These layers can include any of the metallic or polymeric materials disclosed herein, such as electrode layers (e.g., anode and cathode) and semiconducting layers (e.g., organic, inorganic, or hybrid). In certain embodiments, the device is a photovoltaic transducer formed by depositing a first electrode (anode), an active layer (comprising donor and acceptor materials), and a second electrode (cathode), as described above.

[0187] Patterning and Defining Subcellular Dimensions using photolithographic and dry7etching techniques. In some embodiments, reactive ion etching (RIE) or ion beam milling may be employed to achieve lateral dimensions ranging from about 5 nm to 5 mm, as well as sub-ranges or specific values therein. Releasing the Nanoelectronic Devices by removing the sacrificial layer. In some instances, the release is performed by exposing the sacrificial layer to an appropriate chemical etchant, thereby separating the subcellular sized devices from the substrate. Figure 6A-F shows a non-limiting representation of the above method. The formation of a plurality of devices by the above method can include one or more steps of photopatteming, photomasking (by depositing a photoresist), ion-beam milling, and etching (such as with plasma or SF6), as described in detail in Example 1.

[0188] Collecting the Released Devices using a filtration process, centrifugation, or other suitable harvesting techniques. In certain embodiments, filtration is carried out through membranes or filters with pore sizes adapted to capture devices in the sub micrometer range.

[0189] In some instances, for planar devices, the total thickness of the device layers is in the range of about 1 nm to 500 um as well as sub ranges or individual values contained therein. Although described with reference to certain metallic and polymeric layers, it is understood that the fabrication approach is not limited thereto and may be adapted to a variety of materials, including bottom-up grown nanostructures (e.g., vapor-liquid-solid nanowires), solution processed quantum dots or 2D materials, or other hybrid configurations.

[0190] In some instances, the above methods include a further step of forming or adding at least one nanoelectronic circuit onto the nanoelectronic device prior to release from the substrate and collection. Methods of forming nanoelectronic circuits include, for example, CMOS (BEOL) for integrating nanoelectronic circuitry for added functionality in the nanoelectronic devices formed.

[0191] In some instances, one may control the desired dimensions and thicknesses thereof to afford nanoelectronic devices where the dimensions of the nanoelectronic device are about equal or smaller than the diameter of a biological cell and where the nanoelectronic device is capable of transport through vasculature of a subject’s circulatory system. b. Preparation of Bioelectronic Hybrids

[0192] Typically, the nanoelectronic devices described are associated to a biological agent and / or chemical agent which allows the devices to recognize and self-implant at a target region in a body. In some instances, the biological agent is a biological cell, a virus, a cell membrane (or component thereof), an extracellular vesicle (or component thereof), a synthetic cell, and / or a biomolecule thereby forming a bioelectronic hybrid. In one non-limiting instance, a method of forming a bioelectronic hybrid Includes the steps of: (a’) functionalizing a surface of a biological agent with at least one reactive functional group; (b’) functionalizing a surface of a nanoelectronic device, as described herein, with at least one complementary reactive functional group; and (c’) associating the functionalized nanoelectronic device to the functionalized biological agent by reaction of the at least one reactive functional group of the biological agent and the at least one complementary reactive functional group of the nanoelectronic device.

[0193] In some instances, the step of associating involves incubating a plurality of functionalized nanoelectronic devices at a concentration of, for example, 106 devices / mL with functionalized biological agent (for example, at a concentration of 2-4 million / mL) for at least about 1 or 2 hours, or longer. In such instances, the biological agent can be biological cells.

[0194] It is understood that the above method may be used to react a plurality of functionalized biological agents and a plurality of functionalized nanoelectronic devices. The above method produces bioelectronic hybrids having the nanoelectronic device associated to the biological agent, such as a biological cell, as shown in Figure 1C. In most instances, associating involves formation of a covalent link by way of the reaction between the at least one reactive functional group of the biological agent and the at least one complementary reactive functional group of the nanoelectronic device. In one non-limiting instance, the at least one reactive functional group is or includes an azide group and the at least one complementary' reactive functional group is or includes a dibenzocyclooctyne (DBCO) group, which undergo a click reaction to form a covalent link between the groups. Other complementary' sets of reactive groups known in the art can be used to form covalent, non-covalent, ionic, or van der Waals type interactions to form the hybrids. In some instances, the association formed can be an ionic associating, such as when the at least one reactive functional group of the biological agent and the at least one complementary reactive functional group of the nanoelectronic device form a ionic bond pairing. In still other instances, the association formed can be non-covalent association, such as when the at least one reactive functional group of the biological agent and the at least one complementary reactive functional group of the nanoelectronic device form a hydrogen bond pairing.

[0195] In another non-limiting instance, a method of forming a hybrid includes the steps of: (a”) functionalizing or providing a chemical agent having at least one reactive functional group thereon; (b”) functionalizing a surface of a nanoelectronic device, as described herein, with at least one complementary reactive functional group; and (c”) associating the functionalized nanoelectronic device to the functionalized biological agent by reaction of the at least one reactive functional group of the chemical agent and the at least one complementary reactive functional group of the nanoelectronic device.

[0196] Forming the hybrids includes selecting suitable reagents, reaction solvents, reaction temperature, reaction conditions, work up conditions, any needed purification conditions, and hybrid device collection conditions, etc. Different parameters, such as the effects of substrate, growth temperature, pressure etc. can be used to vary the parameters of the device heterostructures formed according to the above methods. c. Characterization of Nanoelectronic Devices and Bioelectronic Hybrids Thereof

[0197] The nanoelectronic devices, and bioelectronic hybrids thereof, can be characterized by various techniques. Without limitation techniques can include use of a probe station connected to a potentiostat for measuring electrical properties; near-infrared (NIR) light transmittance measurements, fluorescence imaging, scanning electron microscopy, focused ion beam - scanning electron microscopy (FIB-SEM), confocal microscopy, metabolic activity assays, and combinations thereof.

[0198] IV. Methods of Using the Nanoelectronic Device and Bioelectronic Hybrids Thereof

[0199] The nanoelectronic devices and bioelectronic hybrids thereof can be used for various applications.

[0200] In one non-limiting instance, a method of providing stimulation and / or modulation to one or more target sites or target regions in a subject includes the steps of: (a'’) administering a pharmaceutical composition including nanoelectronic devices and / or bioelectronic hybrids thereof, as described herein, to a subject; (b’?) allowing the plurality of the nanoelectronic devices and / or bioelectronic hybrids thereof to the arrive at a target site or target region in the subject; and (c'’) applying an externally applied stimulus to the plurality of the nanoelectronic devices and / or bioelectronic hybrids thereof which transduce at least a portion of the externally applied stimulus into energy. Such energy may be, without limitation, electrical energy, mechanical energy, thermal energy, and / or electro-chemical energy.

[0201] In some embodiments, the administering step is performed by intravenous injection or through other routes described above. In some instances, the pharmaceutical composition includes 1 - 100.000,000 nanoelectronic devices per milliliter of total volume of the pharmaceutical composition.

[0202] In some instances of the method, the externally applied stimulus is selected from an optical, electrical, electromagnetic, magnetic, radiofrequency, and / or acoustic stimulus. In certain instances, the externally applied stimulus is applied in a continuous or pulsed mode. In some instances, the externally applied stimulus is applied for at least about 50 ps up to 30 minutes.

[0203] In certain instances, the external energy harvesting transducer of the nanoelectronic devices or bioelectronic hybrids is a photovoltaic transducer and the externally applied stimulus is light, such as near infra-red light. In some instances, the light applied during step (iii) includes at least one of the following: an incident light power of at least about 1 mW / mm2 to about IW / mm2; a pulse width of at least about 1 ps up to about 500 ms: and / or a frequency of at least about 0.01 Hz to about 500 Hz.

[0204] In certain instances, where the nanoelectronic devices are based on a photovoltaic transducer and the externally applied stimulus is optical and may be delivered to the devices, or bioelectronic hybrids thereof, from an optical source, such as a laser source or other source which can generate the desired wavelength(s) needed to generate the electrical energy when stimulating the photovoltaic transducer.

[0205] In certain other instances, the external energy harvesting transducer of the nanoelectronic devices or bioelectronic hybrids is a magnetostrictive, piezoelectric or magnetoelectric transducer and the externally applied stimulus is a magnetic field, acoustic field or electromagnetic field. In some instances, the magnetic field applied during step (hi) includes at least one of the following: a strength of about 0. 1 to 3000 Oe; a frequency of at least about 1 Hz to about 500 Hz; and / or a pulse length of about Ips to 500 ms.

[0206] In certain instances, where the nanoelectronic devices are based on a magnetoelectric transducer and the externally applied stimulus is magnetic and may be delivered to the devices, or bioelectronic hybrids thereof, from a magnetic or electromagnetic source, such as a transmitting coil, AC or DC magnet, or other magnetic field generating device, which can generate the desired magnetic field(s) needed to generate the electrical energy7when stimulating the magnetoelectric transducer. An example transmitter setup is shown in Figure 21.

[0207] The energy transduced / generated by the plurality of nanoelectronic devices or bioelectronic hybrids, when exposed to the externally applied stimulus, provides stimulation or modulation to the target site or target region. In some instances, the stimulation is electrical stimulation which can be used to treat or ameliorate a condition or disease of the subject. In some instances, the target site or target region is associated with a disease or injury selected from the group consisting of Alzheimer's disease, multiple sclerosis, ischemic stroke, different cancers including brain tumor, chronic pain, spinal cord and peripheral nerve injury, paralysis, essential tremor, movement disorders, along with mental disorders such as depression / anxiety, schizophrenia, PTSD, autism, Bipolar Disorder (BPD), Disruptive Mood Dysregulation Disorder (DMDD), cardiovascular disease, autoimmune disease, arthritis, an infection, and combinations thereof. In some cases, the target site or target region is inflamed. In some instances, the target site or target region is in the brain, spine, spinal cord, DRG, peripheral nervous system (PNS), heart, lungs, or kidney. In certain instances, the target site or target region is a site or region in a brain, spinal cord or peripheral nervous system and the electrical stimulation leads to the excitation or inhibition of neurons in the region. In some instances, the modulation includes any of CNS stimulation, PNS stimulation, immune system stimulation, heart stimulation, wound healing and tissue regeneration or tumor treatment.

[0208] In certain instances of the method, the modulation is modulation of biological functions at the one or more target sites or target regions, where the modulation can be selected from electrical, mechanical, thermal, electro-chemical, chemical, and / or optical modulation.

[0209] In some instances of the method, the hybrid bioelectronic devices of the plurality include and can perform at least one functionality selected from the group consisting of sensing, computing, analysis, feedback, and combinations thereof. In some instances, hybrid bioelectronic devices of the plurality7are also capable of wirelessly receiving and / or transmitting commands and / or data. The hybrid bioelectronic devices of the plurality7may be capable of brain-computer interfacing. a. Characterization of Implanted Nanoelectronic Devices and Bioelectronic

[0210] Hybrids Thereof

[0211] The implanted nanoelectronic devices, and bioelectronic hybrids thereof, can be characterized by various techniques. Without limitation, techniques may include imaging, biodistribution studies, microscopy, cytotoxicity7studies, hematoxylin and eosin (H&E) staining. c-Fos positive cell counting, in vivo studies, ex vivo studies, and combinations thereof.

[0212] V. Kits

[0213] The nanoelectronic devices and bioelectronic hybrids thereof can each independently be provided as part of a kit. The (sterile) kit is for medical use. The kit may include instructions for preparing a pharmaceutical composition containing the devices and / or hybrids. The kits may include pharmaceutically acceptable carriers / diluents and other excipients for formulating a pharmaceutical composition of the devices or hybrids.

[0214] In some instances, the kit includes only the nanoelectronic devices and reagents for associating a biological agent and / or chemical agent thereto, as well as instructions for associating a biological / chemical agent to the devices. The biological / chemical agents may be provided with, or separately from, the kit.

[0215] In some instances, the kits may include a syringe or catheter for administration of a pharmaceutical composition containing the devices and / or hybrids. In some instances, the kits may include one or more sources for supplying an externally applied stimulus or stimuli to nanoelectronic devices or bioelectronic hybrids thereof.

[0216] Details of the nanoelectronic devices and bioelectronic hybrids thereof, as well as biological agents, chemical agents, carriers / diluents, excipients, and other agents which may be present in a pharmaceutical composition of a kit, are provided above. The present invention will be further understood by reference to the following nonlimiting examples.

[0217] Example 1: Non-surgical Bioelectronic Implant for Targeted Focal Brain Stimulation using an Optical Stimulus

[0218] Methods:

[0219] 1. Device Fabrication

[0220] Devices were fabricated on two substrates - a 4"’ silicon wafer (p-doped, <100>, singleside polished) and indium-tin-oxide (ITO) coated glass substrate. Silicon wafers were used as purchased from University Wafers and a sacrificial layer of titanium / aluminum (100 nm / 200 nm) was deposited using electron-beam evaporation. Thereafter, the wafers were plasma treated (oxygen, 150 W, 60 seconds) to improve the wettability for a spin-coating process. The ITO coated glass substrates were bought from Sigma and cleaned by sonicating in de-ionized (DI) water, acetone and iso-propyl alcohol (IP A) respectively for 10 minutes each and plasma-treated (oxygen, 150 W, 60 s) before spin-coating.

[0221] Poly(3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS, Ossila) was used as purchased. 5% dimethyl sulfoxide (DMSO. Sigma - to increase the conductivity), 0.2% (3- glycidyloxypropyl)trimethoxy silane (GOPS, Sigma - for higher water stability) and 0.2% Dynol™ 604 (to improve the wettability of the solution while spin-coating) were added to the PEDOT:PSS solution and stirred for 30 mins before use. After mixing, the solution was filtered through a 0.2 pm polyethersulfone (PES) syringe filter. Thereafter, this solution was spin-coated (1000 rpm. 60 seconds) on the substrates (silicon or ITO coated glass substrate) and annealed at 120 °C for 10 minutes.

[0222] The preparation of the organic semiconducting blends P3HT:PCBM (poly (3- hexylthiophene): [6,6]-phenyl-C61-butyric acid methyl ester) or PCPDTBT:PCBM (poly[2,6- (4,4-bis-(2-ethylhexyl)-4H-cyclopenta[2,l-b;3,4-b']dithiophene)-alt-4,7(2,l,3- benzothiadiazole)] : 16.6 |-phenyl-C61 -butyric acid methyl ester) was performed in a glovebox under nitrogen atmosphere. The binary blends were prepared at least 6 hours before spin-coating them on the Si wafers and ITO coated glass substrates. P3HT:PCBM was prepared in a ratio of 1:0.83 with the overall concentration of 22 mg / ml. Similarly, PCPDTBT:PCBM was prepared in a ratio of 1 :2 with the overall concentration of 22 mg / ml. While P3HT:PCBM was spin-coated at 800 rpm (100 ± 12 nm), PCPDTBT:PCBM was spin-coated at 1200 rpm (80 ± 15 nm) for 60 seconds (only one polymer blend at a time) on top of PEDOT:PSS and annealed at 130 °C for 30 minutes. Once the spin-coating was done, the samples were transferred to the electron-beam evaporation machine for deposition of the top metal electrode (titanium (Ti) - 50 nm, 1 A / s). For samples including titanium nitride (TiN) (to provide an improved electrode-electrolyte interface), it was deposited after Ti deposition using reactive sputtering (30 nm, 0.27 A / s, argon:nitrogen = 3: 1 ratio). Once Ti / TiN was deposited, wafers were kept under vacuum until the lithography process.

[0223] For photolithography, the samples were dehydrated at 100 °C for 30 mins to increase the adhesion of the photoresist (AZ3312). Photoresist was spin-coated (3000 rpm, 60 s) on the respective substrates, pre-baked at 100 °C for 60 seconds and exposed using MLA150 (Heidelberg, dose: 130 mJ / cm2at 375 nm). The samples were post-baked at 100 °C for 60 s and then developed using AZ300-MIF developer for 60 seconds. After developing the photoresist, samples were hard baked on a hotplate for 30 minutes at 100 °C.

[0224] For patterning the devices, the substrates were then transferred for dry etching of Ti / TiN and the organic polymers in an Oxford-100 reactive ion etcher. Ti / TiN was etched using SFe plasma (200 W, 3.5 minutes). After Ti / TiN etch, the samples were sonicated in acetone for 5 mins to remove the photoresist and its residuals. After this, oxygen plasma was used to etch away the organic polymers as well as the left-over photoresist (Ti / TiN served as a hard mask for this step). The wafer was cleaved into small chips (20 mm x 20 mm) and stored under nitrogen environment for further use.

[0225] 2 Device Characterization

[0226] A custom-built probe station was used for characterizing the devices (200 pm - 5 pm in size). External laser sources (50 mW, 520 nm - Thorlabs; 100 mW, 785 nm - Coherent) were coupled into the probe station for providing a bottom illumination onto the devices from the PEDOT:PSS side. For the ex-vivo measurements (792 nm laser - HJ Optronics was used) the light passed through the brain before being incident on the devices. The micromanipulators were connected to a potentiostat (CompactStat, Ivium Technologies) for measuring the currentvoltage curves. An upright optical microscope was used to locate the devices on the substrates and aligned with the bottom light source each time before recording the measurements.

[0227] 3. Device Releasing and Collection

[0228] The devices fabricated on the silicon wafers had to be released to create free floating devices. For this, the wafer was cleaved into smaller chips and put inside a 5 mL glass vial. 1.5 ml of diluted tetramethylammonium hydroxide (TMAH) solution (2.7% v / v) was used for a 20 mm x 20 mm silicon chip. The glass vial was then put in a sonication bath for 10 minutes to etch away the sacrificial aluminum layer and collect the devices. Thereafter, the devices were rinsed multiple times (using a custom vacuum-based filtration setup) in DI water to get rid of trace amounts of TMAH. The devices were collected and stored for further experiments (Figure 8A).

[0229] 4. Creation of Cell-Electronics Hybrids: Functionalization, Attachment and Imaging

[0230] Subcellular sized wireless electronic devices (SWEDs) functionalization: SWEDs (5-10 million at a concentration of ~10 million / mL) were immersed in a 10% v / v (3- aminopropyl)tri ethoxy silane (APTES) solution (Sigma) in 100% ethanol overnight at room temperature. Then, these SWEDs were annealed for 2 hours at 75 °C to promote the crosslinking of the APTES molecules on PEDOT:PSS. Then, they were rinsed multiple times in DI water to remove loosely physiosorbed APTES molecules. After this, the APTES-functionalized SWEDs were incubated with equimolar solution of N-succinimidyl 4-[(5-aza-3, 4:7,8- dibenzocyclooct-l-yne)-5-yl]-4-oxobutyrate (NHS-DBCO, Broadpharma) (1 mM solution in phosphate-buffered saline- IX (PBS- IX)) and NHS-Cy3 (ImM, lumiprobe) for 2 hours at room temperature. The SWEDs were constantly stirred during the incubation periods. After the reaction was completed, the SWEDs were rinsed multiple times in PBS-1X and stored at 4 °C till further use.

[0231] Cell Functionalization'. Wehi-265. 1 cells (ATCC) (10-20 million at a concentration of ~2 million / mL) were incubated with succinimidyl 2-azidoacetate (NHS-Azide, Thermo Scientific™) (100 pM in complete cell medium - Dulbecco’s Modified Eagle Medium (DMEM, Sigma) + 10% fetal bovine serum (FBS, ATCC) + 1% penicillin streptomycin (PS. sigma)) solution for 2 hours at 37 °C. Thereafter, the cells were rinsed and plated in complete cell medium.

[0232] Cell-Electronics Hybrids Creation'. After the cell functionalization process, the functionalized SWEDs (5-10 million) were incubated with the cells (10-20 million) in complete cell medium (5 mL) to allow the attachment of the SWEDs to the cell membrane for 2 hours at 37 °C. Periodic agitation was provided to ensure that the SWEDs do not settle down in the petri dish and to increase the SWED attachment efficiency. For experiments requiring the cells to be fluorescent, NHS-Cy5 (100 pM. Lumiprobe) and Qtracker™ 705 (Invitrogen™) were added after incubating SWEDs with the cells for 2 hours and allowed to stain the cells according to the manufacturer protocol.

[0233] Cell-electronics hybrids sorting'. Cell capture and isolation were performed using fluorescence-activated cell sorting (FACS). After incubation, a single-cell suspension was created by filtering the solution from a 35 pm nylon mesh and stored on ice. The single cells were loaded onto the sorter (Sony MA900-1) and passed as a stream in droplets, in front of a laser. Cells with SWEDs attached to them showed higher scattering and fluorescence signal (SWEDs were functionalized with NHS-Cy3), and a double gating strategy was employed to isolate the cell-electronics hybrids with a purity of 92.4% ± 5.2%.

[0234] Fixing and Embedding for Imaging'. The cells attached with the SWEDs were fixed and embedded on a 35 mm glass-bottom petri dish (#1.5). Note that the cells were fixed for imaging purposes only. Live cells were used for intravenous injection. For fixing the cells, 4% paraformaldehyde (PF A. EMS) was added to cells suspended in PBS-1X (cell concentration = 2 million / mL) in equal volume (to make the final concentration of 1 million / mL) and incubated for 10 minutes at room temperature. Thereafter, the fixed suspended cells were rinsed with PBS- IX (at least three times) to remove any fixing reagent from the solution. The rinsed cells were plated on a poly-L-lysine (PLL, Sigma) coated 35 mm glass-bottom petri dish to make them adherent to the glass surface for at least 24 hours. Once the cells had adhered to the surface, the PBS- IX solution was aspirated, and the cells were embedded in agarose (5% w / v in DI water) solution before imaging.

[0235] Imaging'. The cell-electronics hybrids were imaged using a Zeiss Crossbeam 540 SEM / FIB with a serial-sectioning done at 20 nm resolution (Figure 1C). FV1200 Olympus confocal microscope was used for fluorescent imaging. Cells were stained with FAM-DBCO dye and SWEDs showed auto-fluorescence owing to P3HT molecules. Z-stack images of the hybrids were taken simultaneously to confirm the SWED attachment to the cell (not shown).

[0236] 5. Cytotoxicity Assay for Monocytes

[0237] Colorimetric MTT (3-(4.5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays were performed in 96 well plates where SWEDs at various concentrations (10-10000 SWEDs / pL) were added to each well plated with the cells (2-4 million / mL). After each time interval (Figure 9), 10 pL of MTT was added to the medium and incubated for 4 hours. Then 200 pL of DMSO was added and mixed in each well. The absorbance signal was measured using a spectrophotometer (Spark, Tecan) at 570 nm and the background (measured at 630 nm) was subtracted from the signal to obtain the normalized values.

[0238] 6. Subjects Used for Animal Experiments

[0239] Male / female Balb / C mice (Taconic) aged 7-9 weeks were maintained with a 12-hour light / dark cycle and provided food and water ad libitum, also during the duration of the experiment. Animal husbandly' and all experimental procedures were approved by the Massachusetts Institute of Technology' Committee on Animal Care.

[0240] 7. Stereotactic Injection for Inflammation Model

[0241] Note that here the stereotactic injection was done only to create the inflammation model. In actual applications, inflammation will already be present in the target diseased brain regions and use of the devices would not require any surgery'. All mice surgeries were performed under aseptic conditions on a stereotaxic frame. Mice were anesthetized using isoflurane (1 to 4%). Analgesics were provided after anesthesia - subcutaneous injection of buprenorphine sustained release (1.0 mg / kg) before surgery and lidocaine (0.5%, 2-4 mg / kg) was injected subcutaneously at the incision site prior to making the incision. Coordinates used for the intracranial (i.c.) injection into the ventrolateral thalamic nucleus (VL) region relative to bregma were established according to the Allen Brain Atlas2 as follows: anterior-posterior (AP): -1.6 mm. medial-lateral (ML): +1.0 mm, dorsal-ventral (DV): -3.5 mm. Dental drill was used to create an opening in the skull and 1.0 pL of fluorescein isothiocyanate (FITC) conjugated lipopolysaccharide (LPS - 5 pg / L) was injected into the target region. 10 pL nanofil syringe with a 33-gauge beveled needle was used for the injection. Injection speed of 100 nL / min was maintained using a micro-syringe pump and its controller. The syringe was left positioned for 10 mins inside the brain prior to injecting LPS as well as before withdrawal from the brain. Skin tissue was closed with an adhesive and sutures, and the mouse was allowed to recover on a heat pad.

[0242] 8. Intravenous Injection

[0243] After the LPS injection, the mice were allowed to recover on a heating pad and transferred to a cage before the next set of experiments. The cell-electronics hybrids solution was prepared using FACS in the meantime for delivering them via intravenous (i.v. - retro- orbital) injection. Cells were thereafter stained with NHS-Cy5 / QT705 according to the manufacturer’s protocol. The sorted cell-electronics hybrids were then rinsed and re-suspended in PBS at a concentration of 20 million / mL. The final volume of the injection was kept at 100 pL. The entire process was done under sterile conditions. For retro-orbital injection, mice were anesthetized using isoflurane (1 to 4%) and while keeping the mice tilted at an angle (to ensure that the eye-socket plane is perpendicular to the line of the injection), the prepared solution was injected 6 hours after the LPS injection. The mice were allowed to recover after that and kept under observation until they w ere euthanized.

[0244] 9. Perfusion and Imaging

[0245] Anesthetized mice were perfused transcardially with 4% PFA in PBS-1X 72 hours after the retro-orbital injection. Brains were harvested and stored in 4% PFA overnight. 50 pm thick sections of the brain were sliced coronally using a vibratome (Leica VT1000 S) and imaged using a confocal microscope (Nikon 1AR Ultra-fast confocal microscope) for locating the LPS injection site as well as the distribution of the cell-electronics hybrids. LPS w as conjugated with FITC-dye and the cells were stained with NHS-Cy5 / QT705 to allow for their simultaneous imaging in separate fluorescent channels. 10. Biodistribution Study

[0246] The quantification of cell-electronics hybrids was carried out by quantifying the content of Ti (as the SWEDs contain Ti layers) using inductively coupled plasma-mass spectrometry (ICP-MS, Agilent 7900). Specifically, the brain was digested in nitric acid (70%, trace-metal grade) / hydrogen peroxide (30%, trace-metal grade) in a ratio of 5: 1 using UltraWave microwave digester (Milestone). The entire solution was then diluted with milli-Q water to a final concentration of 3% v / v of nitric acid. A blank and six standards (0, 1 ppb to 100 ppm in steps of lOx) were used for calibration of Ti concentration and 103Rh was used as the internal standard during these measurements.

[0247] To quantify the number of implanted SWEDs (PEDOT:PSS|PCPDTBT:PCBM|Ti), the Ti content corresponding to a single SWED was calculated. This was based on the amount of Ti present in the SWED (50 nm Ti deposited on a 10 pm diameter device using electron beam deposition tool). The intrinsic baseline Ti content in brain for control animals (without undergoing any administration of SWEDs) was subtracted from the Ti content in brain for the experimental animals and the result was divided by the Ti content of a single SWED to estimate the number of implanted SWEDs.

[0248] 11. Assessment of Inflammation Influence on Cell-Electronics Localization

[0249] A two-channel imaging system was employed to investigate the localization of selfimplanted cell-electronics hybrids in relation to inflammation, indicated by LPS image intensity using a logistic regression model (P(1|X) = 1 / exp(-01X + 0)). Regions of interest (ROIs) were identified where the cell-electronics hybrids were located. For the actual data from our experiment, 50% of pixels from these ROIs were randomly selected and mapped to corresponding LPS intensities correlated to the inflammation. An equivalent number of pixels from the remaining region, without identified cell-electronics, were randomly selected to collectively create the training dataset. Similarly, the rest of the untrained dataset (50% of pixels from the identified ROIs and an equivalent number of pixels from the remaining region, without identified cell-electronics and not selected in training dataset) were used to create the prediction dataset. For the prediction, a threshold of 0.5 was used for the classification. The final values of the logistic regression analysis (01, 00 and Accuracy (ACC)) were obtained by averaging the results from 1000 repetitions of the fitting for each image.

[0250] In the control, the mapping between the LPS intensity values and the regions with or without identified cell-electronics was shuffled. A logistic regression model was then used to evaluate the influence of inflammation on the localization of cell-electronics, with the beta coefficient (P 1 ) and ACC values to assess the strength of this relationship and the fit of the model, respectively.

[0251] To visually compare the experimental and predicted localizations of cell-electronics hybrids, predicted images were generated (not shown). The experimental data included an image of FITC-conjugated LPS and an image showing the distribution of self-implanted cellelectronics hybrids. The predicted image was generated using the trained logistic regression model, showing the probability of cell-electronics localization based on the LPS intensity.

[0252] 12. Cytotoxicity Assay for Cultured Neurons

[0253] El 8 Sprague Dawley’ Rat dissociated hippocampal neurons w ere purchased from Brainbits. Neurons were cultured in the 96 well plates. The well-plates were coated with 50 pl of poly-D-lysine (100 ug / mL) to promote cell adhesion. Neurons were plated at a concentration of 104 / well and allowed to grow for 7-10 days. 50 pl of SWEDs at various concentrations (105 to 107 mL-1) w ere incubated for the studied time intervals (Figure 11). Thereafter, 10 pL of MTT was added to the medium and incubated for 4 hours. Then 200 pL of DMSO was added and mixed in each well. The absorbance signal was measured using a spectrophotometer (Spark, Tecan) at 570 nm and the background (measured at 630 nm) was subtracted from the signal to obtain the normalized values.

[0254] 13. Testing Immunoreaction to SWEDs

[0255] To examine potential immunoreaction to SWEDs, eliminating the immune reactions stemming from LPS (which is utilized to create the inflammation model), SWEDs were directly injected intracranially into the brain (instead of the self-implantation procedure). Intracranial injection of PBS under identical conditions w as used as a control. SWEDs (at a concentration of 10 million / mL) or PBS (control) w ere unilaterally injected (2pL) using a glass pipette into the mouse brain at the following coordinates - AP: -1.6mm, ML: 1mm. and DV: -2.5mm. Mice were sacrificed at 1, 3, and 7days post-injection (n=3 for each time point). Brains were extracted, coronally sectioned (50pm), and immunohistochemically (IHC) stained for astrocyte marker - glial fibrillary' acidic protein (GFAP) (1 :500, Thermo Fisher) and microglia marker - ionized calcium-binding adapter molecule 1 (Iba-1) (1:500, SYSY) following manufacturer’s protocol. GFAP and Iba-1 IHC used goat anti-rat Alexa Fluor 488 (1 :500, Biotium) and donkey antiguinea pig Alexa Fluor 633 (1 :500, Biotium) respectively' for secondary antibody staining. Hoechst (1 : 10,000, Thermo Fisher) was used to counterstain labeled cell nuclei.

[0256] Confocal imaging of stained sections was done using a Nikon Ti microscope (CSU-X1 confocal module). The SWEDs or PBS injection sites were delineated using a MATLAB script.

[0257] Cell Profiler was used to quantify GFAP and Iba-1 expression based on fluorescence intensity within the defined injection region. Measurements were averaged across 3 sections / animal. SWEDs and PBS groups were statistically compared at each time point.

[0258] 14. Hematoxylin and Eosin (H&E) Staining

[0259] To understand whether the SWEDs cause any tissue damage, (eliminating any effects due to LPS which is employed to create the inflammation model), SWEDs (10 pm diameter, 5 pl solution at a concentration of 10 million SWEDs / mL) were directly injected stereotactically into the hippocampal region (AP: -2mm. ML: 1mm. DV: -2mm). The animals were euthanized via transcardial perfusion in 4% v / v of PFA in PBS-1X at 1-day, 1-week, 1-month and 6-months post-injection. The brain was isolated and stored in 4% PFA (in PBS-1X) overnight before slicing it using a vibratome (Leica VT1000 S). Automated Stainer (Tissue-Tek Prisma Plus) was used for staining the samples with H&E and slides were imaged using a digital whole slide scanner (Aperio) at 40x (not shown).

[0260] 15. In-vivo Stimulation and Analysis

[0261] Inflammation model was created in the animals (n = 9 mice) at the deep brain region (VL) (as described before). All these animals received intravenous (retro-orbital) injection of the cell-electronics hybrids (SWED structure - PEDOT:PSS|PCPDTBT:PCBM|Ti|TiN, 10 pm in diameter) to achieve self-implantation at the inflamed region. The mice were allowed to recover and stimulation was carried out in 4 animals (randomly selected) after 72 hours of the intravenous injection. For carrying out optical stimulation, the mice were anesthetized as previously described while keeping their head fixed. A 792 nm wavelength laser (792 nm - HJ Optronics) was used for stimulation with the following pulse sequence - 15 mW / mm2, 10 ms pulse width, 20 Hz for 20 minutes. These animals were kept anesthetized for an additional 90 mins after the optical stimulation, to allow sufficient time for c-Fos induction, after which they were sacrificed. Control animals (the remaining 5 animals) did not receive the external optical stimulation sequence but underwent the same housing and habituation conditions as the experimental animals including the 90 mins anesthetization period, before they were euthanized.

[0262] After perfusion, the brain was extracted and postfixed in 4% PFA overnight for all the animals. Coronal brain sections (50 pm) were prepared using a vibratome (Leica VT1000 S) and stored in PBS-1X. For immunohistochemistry, tissues were incubated overnight with polyclonal rabbit antibodies raised against c-Fos protein (1 :500, ABE457, sigma). c-Fos positive cells were visualized using immunofluorescence with donkey anti-rabbit Alexa Fluor 647 (1 :500, Thermo Fisher - 2 hours incubation at room temperature). All the cell nuclei were stained with Hoechst (Invitrogen). The stained slices were imaged using a slide scanner (TissueFAXS SL) and a confocal microscope (Nikon Ti, CSU-X1 confocal module) with a 40X objective. c-Fos positive cell counting: To quantify c-Fos positive cells, an initial mask was drawn by a researcher who was blinded to the experiments, to delineate the inflamed region (characterized by nuclei pattern). The mask was then extended by 50 pm in all directions to account for activation of neighboring neurons. Cell Profiler software was utilized to identify all c-Fos positive cells within the masked region, and results were expressed as cells per square millimeter (Figure 4B). An adaptive thresholding approach based on the Otsu algorithm was implemented to segment the cells, with a c-Fos positive cell being defined as having a mean intensity of the region of interest 2.25 times greater than the average background of the entire image and confirmed by colocalization with the cell nucleus.

[0263] Radial distribution of c-Fos positive cells: The radial distribution of c-Fos positive cells was analyzed using radial distance measurements from the inflammation boundary, marked by fluorescent LPS. Initial binary mask to delineate the inflammation boundary and usage of Cell Profiler software, with an adaptive Otsu algorithm, were used for cell identification as described previously. Binary masks of vary ing radial distances were generated using MATLAB, and any ROIs from c-Fos positive cells within these regions were counted and reported per unit area. The distribution of these cells was plotted as a function of radial distance (Figure 4C).

[0264] 16. Biphasic Pulse Generator

[0265] Simulations were performed using Cadence Virtuoso and Spectre. Experimentally measured current-voltage characteristics of 10 pm diameter P3HT (DI) and PCPDTBT (D2) SWEDs when embedded in the brain tissue at a depth of 0.5 mm and wirelessly controlled with 520 nm and 785 nm illumination, were fitted using third-order polynomials. The polynomial fits were used to model the operation of DI and D2 using a Verilog-A script. MOSFETs (metal- oxide-semiconductor field effect transistors) Ml and M2 were modelled using BSIMSOI 4.7.1 with gate length L = 45 nm, gate width W= 450 nm, buried oxide thickness TBOX= 25 nm, top silicon thickness TSi = 10 nm and gate dielectric thickness Tox = 2 nm, as FDSOI (fully- depleted silicon on insulator) FETs. The CMOS footprint for the designed circuit was < 1 pm2(Figures 13A-B). The load w as modelled based on the electrode-electrolyte impedance of a 2 pm diameter raised Pt-electrodes in PBS-1X3, with series parasitic resistance Rp = 1.02 M.Q and pseudo-capacitance CP = 74.4 pF per electrode. Temperature was set at 37 °C to account for operation of the circuit in-vivo.

[0266] The operating points of MOSFETs Ml and M2 were set self-consistently in the subthreshold regime based on the operating points of the driving SWEDs. When actuated by a 785 nm (520 nm) laser pulse, D2 (DI) developed a larger voltage than DI (D2), which appeared as the gate-source voltage (PGS) for M2 (Ml). For Ml and M2 operating in subthreshold regime, a relatively small difference in VGS resulted in a large difference between their drive currents. Thus. M2 (Ml) was selectively turned on and current flow from D2 (DI) to the load and back through M2 (Ml). We created a negative pulse of 65 ps4, a gap (no illumination) of 50 ps followed by a charge balancing, 190 fis positive pulse by remotely controlling the illumination timings. The frequency of operation was chosen to be 200 Hz.

[0267] Discussion:

[0268] This example relates to nanoelectronic devices, and hybrids thereof, that, upon an intravenous injection, are trafficked through the body’s own circulatory system (which is ubiquitous and has continuous and efficient access to all regions) and are self-implanted by recognizing a target brain region, circumventing the need for any surgery. It is further demonstrated that these devices allow for wirelessly controlled focal brain stimulation of the target region deep in a rodent brain, leading to a non-surgical brain implant for targeted focal neuromodulation.

[0269] Such nanoelectronic devices represent the development of wireless free-floating electronic devices that are miniaturized to fit inside a fluid system of a body; that demonstrate efficient circulation without being eliminated from the bloodstream; and that demonstrate recognition of and self-implantation in the target brain region. To achieve the aforementioned, this example demonstrates wireless optical energy harvesting electronic devices that are subcellular sized and self-standing and hybrids thereof with living immune cells. An exemplary target region was chosen to be a region of inflammation, as it is a hallmark and key therapeutic target of various diseases of the brain, and other nervous systems.

[0270] A discussion of the nanoelectronic device design and their fabrication, as well as the creation of cell-electronics hybrids and their use in non-surgical focal brain stimulation is provided below.

[0271] Subcellular Sized Free-floating Wireless Electronic Devices for Brain Stimulation

[0272] To fit and freely move inside the vasculature without clogging, the size of nanoelectronic devices was chosen to be similar to or smaller than that of the circulating cells (for reference, a circulating cell such as monocyte can have a diameter of 12-18 pm). Hence, subcellular sized wireless electronic devices (SWEDs) were developed that are free-floating and that can convert extracorporeally applied fields to electrical energy to enable electrical neuromodulation. In this example, the photovoltaic principle, which involve wireless powering via optical fields, was selected. However, other t pes of extracorporeally applied fields (such as electrical, radio frequency, magnetic, or acoustic) can be used to generate electrical energy with a suitable nanoelectronic device. In this example instance, optical modalities were chosen for their ability- to provide high spatio-temporal resolution, penetration depth of several centimeters in the human head with intact skull and because they have already been employed for clinical studies. Moreover, photovoltaic devices generate DC potentials (Sze, S., et al., Physics of semiconductor devices. (2021)) eliminating the need for any rectifying circuits (saving on-chip area and avoiding circuit complexity). Other modalities can also be employed based on user defined requirements.

[0273] Though photovoltaics have been applied previously for neuromodulation (Acaron Ledesma, H. et al., Nature Nanotechnology 14, 645-657 (2019)), subcellular sized free-floating photovoltaic devices compatible with circulation through bloodstream have not, to the best of our knowledge, been demonstrated previously for in vivo electrical brain stimulation. Moreover, none of the previous photovoltaic devices have been demonstrated for non-surgical targeted focal brain stimulation.

[0274] Organic semiconductors (Chander, N., et al., Sol. Energy Mater. Sol. Cells 161, 407-415 (2017); and MacDiarmid, A. G. & Epstein, A. J. Makromol. Chemie. Macromol. Symp. 51, 11- 28 (1991)) were employed to leverage the photovoltaic effect, as they have unique advantages such as narrow bandwidth for enabling multiplexing, high optical absorption coefficients, mechanical flexibility allowing good interface with soft biological systems and biocompatibility. They also provide ease of fabrication and compatibility- with Complementary- Metal-Oxide- Semiconductor (CMOS) back-end-of-line (BEOL) processing (Gowrisanker, S. el al., Org. Electron. 10. 1217-1222 (2009); and Baierl, D. et al.. Nat. Commun. 3. 1175 (2012)) creating opportunities for further integrating further functionalities. The equivalent electrical circuit of these photovoltaic devices consists of a current source (representing optical intensify dependent polaron generation), three diodes and several resistors.

[0275] The nanoelectronic devices consisted of a 3-layer structure - anode, binary blend of semiconducting organic polymers (acceptor and donor material forming the active layer where the excitons are generated) and cathode. By customizing the organic polymeric materials, these nanoelectronic devices can be tuned to different optical wavelengths which will allow their independent control, enabling multiplexing. To achieve this, two different donor materials were used, poly (3 -hexylthiophene) (P3HT) and poly[2,6-(4,4-bis-(2-ethylhexyl)-4H-cyclopenta[2,l- b;3,4-b']dithiophene)-alt-4,7(2,l,3-benzothiadiazole)] (PCPDTBT), as their absorption spectra are complimentary- to each other. [6, 6]-phenyl-C61 -butyric acid methyl ester (PCBM) is used as the acceptor polymer in both cases. Further, poly(3,4-ethylenedioxythiophene): polystyrene sulfonate (PEDOTPSS) and titanium (Ti) are used as the anode and cathode respectively and chosen based on their work function (Figure 5) and biocompatibility. To investigate the scaling behavior and gauge the feasibility of sub-cellular sized devices, a fabrication process was developed to create devices with nanoscale thickness (about 200 nm) and different lateral length-scales, ranging from diameters of 200 pm (> lOx the diameter of a monocyte) to 5 pm (subcellular size). The nanoscale thickness provides increased mechanical flexibility for improved biological interfacing but also for tuning device performance due to the competing effects of increase in exciton generation, as well as recombination effects with the increase in device active layer thickness (Min Nam. Y.. et al.. Sol. Energy) Mater. Sol. Cells 94, 1118-1124 (2010)). These devices can be mass produced at the wafer-scale (Figure 6). Scanning Electron Microscopy (SEM) images (Figure IB) demonstrated a multitude of fabricated devices on a 4-inch silicon wafer.

[0276] The set-up for characterizing the nanoelectronic devices is shown in Figure 2A. The scaling performance of the devices is shown in Figure 2B. It is seen that these devices, even when scaled down to diameters <= 10 pm achieving subcellular sizes (and less than O.Olx volume of a cell with 12 pm diameter) can generate nanowatts of pow er. Figure 2C show s the current-voltage characteristics of a SWED (10 pm in diameter) illustrating the increase in short- circuit current (ISC) generation with the incident optical intensity. The open circuit voltage (VOC) does not change significantly even when operating the devices at low intensities which corroborates the ability of photovoltaics to generate near-constant potential in the open circuit condition. Figures 2B and 2C correspond to P3HT based devices and the characteristics of the PCPDTBT based devices are shown in Figures 7A-B. Even at this ultra-small dimension (subcellular sizes), SWEDs generate open-circuit voltages, VOC = 0.2 ± 0.008 V (P3HT), VOC = 0.17 ± 0.01 V (PCPDTBT) and short-circuit currents, ISC = 12.8 ± 2.15 nA (P3HT), ISC = 18.2 ± 2.56 nA (PCPDTBT) at incident optical intensity of 10 mW / mm2

[0277] Next, to achieve free-floating SWEDs a process flow to release the device structures from the fabrication substrate (silicon wafer) was employed through tetramethylammonium hydroxide (TMAH) based etching of a sacrificial aluminum layer in order to retrieve and collect them (Figure 8A). Characterizations confirmed that even after the substrate-release process, the devices retained good performance (Figure 8C).

[0278] To characterize the energy harvested by the SWEDs when implanted in the brain, an ex- vivo measurement setup allowing probing of the devices and measurement of the generated powder was employed, with light penetrating through different brain tissue thicknesses (Figure 2A). The current- voltage characteristics are plotted in Figure 2D for various tissue thicknesses at an incident intensity of 24.6 mW / mm2of near-infrared (NIR) light, showing the successful remote operation of the SWEDs (10 pm in diameter) even for the entire mouse brain with the intact skull. The power generated by the SWEDs is plotted as a function of optical intensity for the whole brain without and with the skull in Figure 2E and as expected the power increases with an increase in the incident optical intensity. It is seen that these SWEDs can generate 0.545±0.058 nW and 0.482±0.019 nW of power (for the whole brain without and with the skull, respectively) at an incident optical intensity of 46.06 mW / mm2(note that 100 mW / mm2of NIR light can be used continuously in brain with an acceptable temperature increase of about 2 °C, while higher intensity can be used when using pulsed mode (Podgorski, K. & Ranganathan, G.

[0279] J. Neurophysiol. 116, 1012-1023 (2016)). The ability to independently control the SWEDs using different optical wavelengths when operated through brain tissue was also demonstrated.

[0280] Autonomous Implantation in the Target Brain Region without External Guidance

[0281] Next, it was investigated whether the SWEDs could be transported through the circulatory system and autonomously implanted (circumventing the need for external guidance) in a target region without any surgery. While electronic devices lack an intrinsic ability to navigate the body’s circulatory system and can be eliminated from the bloodstream by the immune system or subject to biological barriers, biological cells, on the other hand, are complex biological machineries naturally adapted and optimized to perform these types of functions (Li, C.-X., et al., Adv. NanoBiomed Res. 1, 2000052 (2021)). Hence, cells such as erythrocytes, monocytes, platelets, neutrophils, lymphocytes and stem cells, were considered as means for transport to target diseased regions (Li, C.-X., et al., Adv. NanoBiomed Res. 1, 2000052 (2021)).

[0282] It was considered whether cells could be used to transport and install the nanoelectronic devices. To achieve this, cell-electronic hybrids, bioelectronic hybrids, were formed (Figure 1 C) wherein the SWEDs are associated onto a cell surface in order to circulate in the bloodstream and self-implant in specific regions. Based on the target disease of interest, appropriate cells can be selected.

[0283] In this example, an inflamed region in the brain was chosen as the target, as targeted electrical modulation of the region of inflammation can address the inherent mechanisms underly ing a broad spectrum of neurological diseases. For the biological cell, immune cells (ImCs) were selected, more specifically monocytes, as they are known to target region(s) of inflammation (Shi. C. & Pamer, E. G. Nat. Rev. Immunol. 11. 762-774 (2011)). Moreover, monocytes are also able to cross the blood-brain barrier (Marchetti, L. & Engelhardt, B. Vase. Biol. 2, Hl -Hl 8 (2020)).

[0284] To create the bioelectronic hybrids, the surface of monocytes was functionalized with azide groups leveraging the available amines on the cell membrane proteins. On the devices, the surface of the PEDOT:PSS layer of the SWEDs were functionalized with dibenzocyclooctyne (DBCO) groups, so that the SWEDs could be attached to the cells using Click chemistry (Meyer, J. P., et aL. Bioconjug. Chem. 27, 2791-2807 (2016)). Fluorescence-activated cell sorting (FACS) was performed to isolate cell-electronics hybrids with a purity of 92.4% ± 5.2% (Figure 3A).

[0285] The 3D reconstructed Focused Ion Beam-Scanning Electron Microscopy (FIB-SEM) image of the hybrid of a SWED with a monocyte is shown in Figure 1C. To assess whether the SWEDs have any adverse effect on the ImCs. a metabolic activity assay (3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide, MTT) was performed. Results indicated that the health and viability of the monocytes were not impaired by the SWEDs (Figure 9).

[0286] To investigate whether the bioelectronic hybrids could be self-implanted in a target inflamed brain region, a classical inflammation model in a deep brain region (ventrolateral thalamic nucleus (VL)) of a mouse (Balb / C) was created through stereotactic injection of lipopolysaccharide (LPS) (1.0 pL at 5 pg / pL using a 33-gauge beveled needle at an injection speed of 100 nL / min) functionalized with fluorescent dyes, under sterile conditions and isofl urane anesthesia. The dye allowed for precisely locating the region of LPS injection later during postmortem imaging. Note that here the stereotactic surgery was done to emulate the inflammation. In actual applications, as mentioned herein, the diseased brain regions exhibit intrinsic inflammation, and use of the nanoelectronic devices would not require any surgery.

[0287] After the inflammation model was created, high purity suspension of cell-electronics hybrids were injected through intravenous (retro-orbital) injection. 72 hours after administering the intravenous injection, the mice were transcardially perfused, their brains were harvested, fixed, and brain slices were prepared for imaging. It was shown that the bioelectronic hybrids self-implanted in the region of inflammation in the brain (imaging not shown). In one control group, where only SWEDs were intravenously injected without attachment to ImCs with all other conditions remaining same as the experimental animals, SWEDs were not observed in the brain images (imaging not shown), demonstrating the role of ImCs for effective selfimplantation. Another control group w as studied with the same conditions as experimental animals except that LPS was replaced with phosphate-buffered saline (PBS), and no hybrids were detected in the brain images (imaging not shown) corroborating that the cell-electronics hybrids selectively entered inflamed brain regions.

[0288] Biodistribution studies (Figure 10A) w ere performed and quantified the number of SWEDs (10 pm) that were implanted in the brain (Figure 3B) and found that about 14029 ± 4154 (n=3) SWEDs were installed in the inflamed brain of experimental animals. The number of SWEDs implanted in the two control groups was quantified and it was found that the number was insignificant in both cases (Figure 3B). Further, to evaluate the co-localization of selfimplanted cell-electronics hybrids with the target (inflamed) region, logistic regression analysis was employed. The results showed inflammation was a determining factor in predicting the selfimplantation of cell-electronics hybrids. The high coefficient (pi, actual=2.2±0.84; pi,shuffled=l. lxlO-4±6xlO-4; p=0.0019) (Figure 3C) and a high accuracy (ACC) value (ACCactual=0.82±0.05; ACCshuffled=0.50±0.01; p=0.000009) (Figure 3D) of the actual experimental data compared to the shuffled control further confirmed the prediction.

[0289] The biocompatibility of the SWEDs was indicated by an MTT assay conducted on cultured primary neurons (Figure 11) as well as the examination of potential immunoreaction to SWEDs in vivo (glial fibrillary acidic protein (GFAP) and ionized calcium-binding adapter molecule 1 (Iba-1) levels, (Figures 12A-B)) and tissue damage (hematoxylin and eosin (H&E) staining (not shown)).

[0290] The cell-electronics hybrids have also been shown to cross the blood-brain-barrier (Figure 22).

[0291] Targeted Focal Brain Stimulation Employing Non-surgical Bioelectronic Hybrid Implants

[0292] Having established that the SWEDs could self-implant in the target region in the brain, without any surgery, their capabilities for targeted focal brain stimulation were explored. The cellelectronics hybrids were introduced intravenously in a mouse with inflamed deep brain region (VL) (as described in the previous section) and after self-implantation, near infrared (NIR) light with a 792 nm wavelength was used for wireless actuation of the SWEDs ( 15 mW / mm2intensity, 10 ms pulse width, 20 Hz frequency, 20 mins duration) (Figure 4A). A control group involved animals which were processed exactly the same way as above but where no optical pulses were applied. Brain stimulation was characterized using c-Fos immuno-histochemistry, which is widely used to delineate cell activity (Kovacs, K. J. Journal of Neuroendocrinology 20, 665-672 (2008); and Morgan, J. I. & Curran, T. Trends in Neurosciences 12, 459-462 (1989)). Quantification of the brain slices revealed significantly higher number of c-Fos positive cells (315±81 cells / mm2, n=4) in the target (inflamed) deep brain region of the experimental mice compared to the control group (112±40 cells / mm2, n=5) (Figure 4B). The analysis further revealed that c-Fos positive cells were predominantly localized around the inflamed region in the experimental group, a pattern not observed in the control group (Figure 4C). These results demonstrated the capability' of the bioelectronic hybrids to act as a brain stimulator to provide focal stimulation of the targeted brain region. Conclusions:

[0293] A non-surgical bioelectronic implant for the brain was demonstrated that was shown to autonomously implant without external intervention and enable wirelessly controlled, focal brain stimulation of the targeted brain region. The subcellular sized, free-floating nanoelectronic devices detailed in this example were able to travel through the vasculature, recognize and selfimplant in a target region and were wirelessly actuated using optical fields for focal brain stimulation deep in rodent (mouse) brain.

[0294] Example 2: Non-surgical Bioelectronic Implants using Magnetic, Electromagnetic or Acoustic Stimulus

[0295] Core-shell magnetoelectric heterostructures of magnetostrictive and piezoelectric materials can have oscillating magnetic fields applied thereto which will create strain due to the presence of the magnetostrictive core. This strain can be transduced into voltages through the piezoelectric shell component thereby creating electric energy from magnetic fields (Figure 14). Alternatively, magnetostrictive or piezoelectric nanodevices can be used alone to convert magnetic or acoustic stimulus to mechanical energy or electrical energy respectively.

[0296] An exemplary device can employ barium titanate (BaTiOs, BTO) as a piezoelectric shell material, and cobalt ferrite (CoFe2O4, CFO) as a magnetostrictive core material. These materials are compatible with chemical synthesis processes and can be configured into core-shell nanostructures of 200 nm average size. The devices demonstrate significant magnetoelectric coupling coefficients, thus allowing significant electric field generation upon application of low AC magnetic fields.

[0297] The magnetostrictive core can be synthesized through coprecipitation from Iron Nitrate (III) and Cobalt Nitrate (II) precursors under controlled conditions. Different parameters, such as temperature, pH, precursor concentrations, and reaction time can be used to vary the properties of the core structures formed. The piezoelectric shell can be synthesized through a sol-gel process, with careful control over the core-shell ratio to achieve optimal device performance.

[0298] The structural and compositional characteristics of the devices can be studied using transmission electron microscopy (TEM) coupled with energy dispersive X-ray spectroscopy (EDS) (Figures 15A-C). The magnetic properties of the magnetostrictive core can be characterized using vibrating sample magnetometry (VSM), showing saturation magnetization values of approximately 55 emu / g at 10000 Oe (Figure 16). The piezoelectric properties can be characterized using piezoelectric force microscopy (PFM) (Figure 17). The performance of the core-shell magnetoelectric devices can be characterized through measurements of the magnetoelectric (ME) coupling coefficient (Figure 18). Measurements show a maximum CINE of 9 mV / cmOe at 200 mT DC bias when excited by a 1 mT AC magnetic field (100 Hz). The devices are expected to operate effectively at non-resonant frequencies required for electrical stimulation of neurons, as low frequency magnetic fields have excellent penetration through tissue.

[0299] Alternatively, these devices can also be fabricated using conventional top-down microfabrication techniques such as deposition, etching, and substrate release to create free floating structures of different sizes.

[0300] For non-surgical implantation, the devices can be attached to immune cells (IC), specifically monocytes, as they can target regions of inflammation and cross the blood brain barrier (BBB). The cell surface can be functionalized with groups such as Dibenzocyclooctyne (DBCO) leveraging the available amines on the cell membrane proteins. The devices can be functionalized with groups such as azides using appropriate surface chemistry, enabling attachment to the ICs using Click chemistry (Figure 19).

[0301] In an exemplary approach, lipo-polysaccharide (LPS) functionalized with fluorescent dyes can be stereotactically injected (1.0 pL at 5 pg / pL) to create a localized region of inflammation. The fluorescent dye enables precise location tracking of the inflammation site. It should be noted that in actual therapeutic applications, inflammation would be naturally present in diseased brain regions and no invasive surgery would be required.

[0302] Following the creation of the inflammation model (approximately 1-4 hours post injection), the IC-device hybrids can be delivered through retro-orbital injection (100-150 pL at a concentration of approximately 1-100 million devices per mL). The devices can be functionalized with fluorescent molecules to enable visualization of their distribution through immunohistochemistry (Figure 20). Real-time imaging of the devices circulating through the vasculature can be performed using appropriate imaging systems to understand circulation dynamics and timing of brain entry'.

[0303] In some instances, the nanoelectronic devices may be encapsulated to protect them from wet environments. One or more coatings can be used to improve biocompatibility. The effect of different sizes and densities of the devices on neural activation can be examined.

[0304] For actuation of the self-implanted devices, acoustic, electromagnetic or magnetic field generators for example, 3D-printed coil setups with or without ferrite cores can be employed (Figure 21). These setups can be designed to provide the required actuation parameters including magnetic field strengths and operational frequencies for electrical stimulation. EMBODIMENTS

[0305] Some embodiments of the present disclosure can be described in view of one or more of the following:

[0306] Embodiment 1. A hybrid bioelectronic device comprising: a wirelessly actuatable component configured to interact with an externally applied stimulus; and a biological or chemical component attached to the wirelessly actuatable component, wherein the hybrid bioelectronic device is configured to be administered into a body of a subject and thereafter recognize, via the biological or chemical component, a target region within the body and selfimplant at the target region.

[0307] Embodiment 2. The hybrid bioelectronic device of embodiment 1, wherein the wirelessly actuatable component has a size that is between 5 nm and 5 mm.

[0308] Embodiment 3. The hybrid bioelectronic device of Embodiment 1 or 2, wherein the wirelessly actuatable component comprises a transducer configured to convert the externally applied stimulus into electrical, mechanical, thermal, or electro-chemical energy.

[0309] Embodiment 4. The hybrid bioelectronic device of any one of Embodiments 1 to 3, wherein the wirelessly actuatable component further comprises (i) a nanoelectromc circuit configured to be powered by a / the transducer, and / or (ii) an antenna for wireless communication.

[0310] Embodiment 5. The hybrid bioelectronic device of Embodiment 3 or 4, wherein the transducer is optical, magnetostrictive, piezoelectric, or magnetoelectric.

[0311] Embodiment 6. The hybrid bioelectronic device of any one of Embodiments 1 to 5, wherein the externally applied stimulus is an electrical, optical, electromagnetic, magnetic, or acoustic stimulus, or a combination thereof.

[0312] Embodiment 7. The hybrid bioelectronic device of any one of Embodiments 3 to 6, wherein the transducer is a planar structure or a non-planar structure, optionally wherein the non-planar structure is spherical or irregular in shape.

[0313] Embodiment 8. The hybrid bioelectronic device of any one of Embodiments 3 to 7, wherein the transducer is an optical transducer which comprises a substrate-free optoelectronic device.

[0314] Embodiment 9. The hybrid bioelectronic device of Embodiment 8, wherein the substrate-free optoelectronic device consists of an active layer and optionally one or more additional layers selected from a group consisting of anode layers; cathode layers; electron transport layers (ETLs); hole transport layers (HTLs); layers for improving interface properties or stability; protective layers; and combinations thereof. Embodiment 10. The hybrid bioelectronic device of Embodiment 9, wherein the active layer comprises an organic semiconductor.

[0315] Embodiment 11. The hybrid bioelectronic device of Embodiment 9 or 10, wherein the active layer is formed of at least one donor material, at least one acceptor material, or a combination thereof.

[0316] Embodiment 12. The hybrid bioelectronic device of Embodiment 11, wherein the at least one donor material is an organic polymer, optionally wherein the organic polymer comprises poly[(2,6-(4,8-bis(5-(2-ethylhexyl-3-fluoro)thiophen-2-yl)-benzo[l,2-b:4,5- b']dithiophene))-alt-(5,5-(T,3'-di-2-thienyl-5',7'-bis(2-ethylhexyl)benzo[T,2'-c:4',5'- c'] dithiophene-4, 8-dione)] (PM6), poly[(2,6-(4,8-bis(5-(2-ethylhexyl-3-fluoro)thiophen-2-yl)- benzo[l,2-b:4,5-b']dithiophene))-alt-(5,5-(r,3'-di-2-thienyl-5',7'-bis(2-ethylhexyl)benzo[T,2'- c:4',5'-c']dithiophene-4,8-dione)] (DI 8), poly[(4,4'-bis(2-ethylhexyl)dithieno[3,2-b:2',3'- d]silole)-2,6-diyl-alt-(2,l,3-benzothiadiazole)-4,7-diyl] (PSBTBT), poly[N-9'-heptadecanyl-2,7- carbazole-alt-5,5'-(4',7'-di-2-thienyl-2',r,3'-benzothiadiazole)] (PCDTBT), poly(3- hexylthiophene) (P3HT), poly(2,6-(4,4-bis-(2-ethylhexyl)-4H-cyclopenta[2,l-b:3,4- b’ |dithiophene)-alt-4.7-(2.1.3-benzolhiadiazole) (PCPDTBT), poly(2,5-thiophene) (P3T), poly(3,4-ethylenedi oxythiophene) (PEDOT), poly(9,9-dioctylfluorene) (PFO), poly(2,5-bis(3- alkylthiophen-2-yl)thieno[3,2-b]thiophene) (PBTTT), poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS), poly(3,6-dioctylthieno[3,2-b]thiophene) (P3OT), poly (2.5- thiophene vinylene) (PTV), polythiophenes with various alkyl side chains and substitutions, poly(3,4-difluorothiophene-co-bithiophene) (PFTB), poly(2,6-carbazole) (PCz), poly(9,9- dioctylfluorene-co-bithiophene) (F8T2), poly (3-hexylthiophene) (P3HT), poly[2,6-(4,4-bis-(2- ethylhexyl)-4H-cyclopenta[2,l-b;3,4-b'] di thiophene)-alt-4, 7(2, 1,3-benzothiadi azole)] (PCPDTBT) or a combination thereof.

[0317] Embodiment 13. The hybrid bioelectronic device of Embodiment 11 or 12, wherein the at least one acceptor material is an organic polymer, optionally wherein the organic polymer comprises L8-BO (5,5'-Bis((2-butyloctyl)oxy)-[l,r:3',l"-terphenyl]-4,4"-dicarbaldehyde), Y6 (2,2'-((2Z,2'Z)-((12,13-bis(2-ethylhexyl)-3,9-diundecyl-12,13-dihydro-[l,2,5]thiadiazolo[3,4- e]thieno[2.3":4',5']thieno[2',3,:4.5]pyrrolo[3,2-g]thieno[2',3':4,5]thieno[3.2-b]indole-2.10- diyl)bis(methanylylidene))bis(5,6-difluoro-3-oxo-2,3-dihydro-lH-indene-2,l- diylidene))dimalononitrile), Z8 (2,2'-((2Z,2'Z)-((12,13-bis(2-ethylhexyl)-3,9-diundecyl-12,13- dihydro-[1.2,5]thiadiazolo[3,4-e]thieno[2,3":4',5']thieno[2',3':4,5]pyrrolo[3,2- g]thieno[2',3':4,5]thieno[3.2-b]indole-2,10-diyl)bis(methanylylidene))bis(3-oxo-2,3-dihydro-lH- indene-5,6-dicarbonitrile))dimalononitrile), o-BTP-eC9 (2,2'-((2Z,2'Z)-((12,13-bis(2- ethylhexyl)-3,9-bis(2-(2-ethoxyethoxy)ethyl)-12,13-dihydro-[l,2,5]thiadiazolo[3,4- e]thieno[2,3":4',5']thieno[2',3,:4,5]pyrrolo[3,2-g]thieno[2',3':4,5]thieno[3.2-b]indole-2,10- diyl)bis(methanylylidene))bis(3-oxo-2,3-dihydro-lH-indene-2,l-diylidene))dimalononitrile), P- 2BTh-F (3,9-bis(2-methylene((3-(l,l-dicyanomethylene)-6,7-difluoro)indanone))-5,5,ll,l l- tetrakis(4-hexylphenyl)dithieno[2,3-d:2',3'-d']sindaceno[l,2-b:5,6-b']dithiophene), and PC70BM ([6.6]-Phenyl-C71 -butyric acid methyl ester, PCBM (Phenyl-C61 -butyric acid methyl ester). ITIC (Indacenodithieno[3.2-b]thiophene-2,8-dicarboximide), IT-M (Indacenodithiophene-based molecule), EH-IDTBR (2,2’-((2Z,2’Z)-((12,13-bis(2-ethylhexyl)-3,9-diundecyl-12,13-dihydro- [l,2,5]thiadiazolo[3,4-e]pyrazine-2,2,6,6-tetrayl))bis(4,5-dihydrothieno[3,4-b][l,4]dioxine-5,7- diyl))bis(methanylylidene))bis(3-ethyl-2-thioxothiazolidin-4-one), [6,6]-phenyl-C61 -butyric acid methyl ester (PCBM) or a combination thereof.

[0318] Embodiment 14. The hybrid bioelectronic device of any one of Embodiments 9 to 13, wherein the anode and cathode layers are electrically conducting materials.

[0319] Embodiment 15. The hybrid bioelectronic device of any one of Embodiments 9 to 14, wherein the ETL layer comprises (i) an organic polymer, optionally wherein the organic polymer poly[[2.7 bis(2 ethylhexyl) 1.2.3.6.7.8 hexahydro 1,3, 6, 8 tetraoxobenzo[lmn]phenanthroline 4,9 diyl] -2,5 thiophenediyl[9,9 bis[3' ((N,N dimethyl)-N ethylamino)propyl]-9H fluorene 2,7 diyl] -2,5 thiophenediyl] (PNDIT F3N); (ii) a metal or metal oxide, optionally wherein the metal oxide comprises zinc oxide (ZnO), titanium dioxide (TiCh); or (iii) a combination of (i) and / or (ii) thereof.

[0320] Embodiment 16. The hybrid bioelectronic device of any one of Embodiments 9 to 15, wherein the HTL layer comprises (i) an organic polymer, optionally wherein the comprises poly(3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS); (ii) a metal or metal oxide, optionally wherein the metal oxide comprises molybdenum trioxide (MoOs), tungsten tri oxide (WO3), vanadium pentoxide (V2O5); or (iii) a combination thereof.

[0321] Embodiment 17. The hybrid bioelectronic device of any one of Embodiments 9 to 16, wherein the lay ers for improving interface properties comprise: (a) metal, metal alloy, or metal compound, optionally wherein (i) the metal or metal alloy comprises platinum (Pt), platinum iridium alloy (Pt Ir), gold (Au), porous Au, tungsten (W). nano Pt, tantalum, or a combination thereof, and / or (ii) the metal compound comprises titanium nitride (TiN), iridium oxide (IrOx), Sputtered Iridium Oxide Film (SIROF), Activated Iridium Oxide Film (AIROF), ruthenium oxide (RuOx), tantalum oxide (Ta2Os), gold oxide (AuOx), platinum oxide (PtOx). palladium oxide (PdOx), or a combination thereof; (b) a PEDOT:PSS or a variant thereof, optionally wherein the variant comprises PEDOT:PSS / Ptlr, a PEDOT / carbon nanotube (CNT) composite, or a combinations thereof; (c) a carbon-based material, optionally wherein the carbon-based material comprises glassy carbon, porous graphene. CNT fibers, graphene fibers, reduced graphene oxide, hybrid diamond / carbon fiber, 3D fuzzy graphene, or a combination thereof; or (d) a combination thereof.

[0322] Embodiment 17. The hybrid bioelectronic device of any one of Embodiments 9 to 16, wherein the layers for improving interface properties comprise: a) metals and metal oxides such as titanium nitride (TiN), platinum (Pt), iridium oxide (IrOx). including Sputtered Iridium Oxide Film (SIROF) and Activated Iridium Oxide Film (AIROF), ruthenium oxide (RuOx), platinum-iridium alloy (Pt-Ir), Tantalum / Ta2O5, gold (Au) and porous Au, oxidized forms such as AuOx, PtOx, or palladium oxide (PdOx), tungsten (W), Nano Pt; b) PEDOT:PSS and variants such as PEDOT:PSS, PEDOT:PSS / Ptlr. PEDOT / CNT (incorporating carbon nanotubes); or c) carbon-based materials such as glassy carbon, porous graphene, carbon nanotube (CNT) fibers, graphene fibers, reduced graphene oxide, hybrid diamond / carbon fiber, 3D fuzzy' graphene, and combinations thereof.

[0323] Embodiment 18. The hybrid bioelectronic device of any one of Embodiments 5 to 17, wherein the transducer is an optical transducer which is based on inorganic materials.

[0324] Embodiment 19. The hybrid bioelectronic device of any one of Embodiments 5 to 18, wherein the transducer is an optical transducer and the optical transducer comprises one or more inorganic semiconductor-based devices, optionally wherein the one or more inorganic semiconductor-based devices comprise silicon nanowires with p-i-n junctions, III-V semiconductor nanowires, heterostructure nanowires, GaAs-based thin film planar devices, crystalline silicon-based thin film plana devices, amorphous silicon-based thin film planar devices, CIGS, -based thin film planar devices, or CdTe-based thin film devices, optionally wherein the optical transducer comprises one or more encapsulating or passivating layers to ensure biocompatibility and stable interfacing with biological medium.

[0325] Embodiment 20. The hybrid bioelectronic device of any one of Embodiments 5 to 19, wherein the transducer is an optical transducer, and the optical transducer is based on porphyrinbased materials.

[0326] Embodiment 21. The hybrid bioelectronic device of any one of Embodiments 5 to 7. wherein the transducer is a magnetoelectric transducer and the magnetoelectric transducer is formed either as a composite of magnetostrictive and piezoelectric materials or as a single-phase material exhibiting intrinsic coupling betw een magnetic and electric properties. Embodiment 22. The hybrid bioelectronic device of Embodiment 2E wherein the magnetoelectric transducer is formed as a composite of magnetostrictive and piezoelectric materials and the composite is either in a core-shell structure or in a planar layered structure.

[0327] Embodiment 23. The hybrid bioelectronic device of any one of Embodiments 1 to 22, wherein the biological agent comprises at least one of biological cells, viruses, cell membranes, extracellular vesicles, synthetics cells, or biomolecules, optionally wherein the biological agent is genetically engineered or biologically or chemically modified.

[0328] Embodiment 24. The hybrid bioelectronic devices of Embodiment 23, wherein the biological cells comprises immune cells (such as monocytes, macrophages, T cells, neutrophils, and Natural Killer cells), stem / progenitor cells (such as MSCs, iPSCs, HSCs. and neural progenitor cells), glial cells (such as Schwann cells), structural cells (such as epithelial cells), or blood cells (such as red blood cells), or combinations thereof.

[0329] Embodiment 25. The hybrid bioelectronic device of any one of Embodiments 1 to 24, wherein the biological or chemical agent is attached to the wirelessly actuatable component by a linker that is permanent, self-degrading, or cleavable by a biological signal or by exposure to an external stimulus.

[0330] Embodiment 26. The hybrid bioelectronic device of any one of Embodiments 1 to 25, wherein the hybrid bioelectronic device is configured to be able to be transported through a fluid system of the body of the subject.

[0331] Embodiment 27. The hybrid bioelectronic device of Embodiment 25, wherein the fluid system comprises a vascular system (such as blood or plasma), lymph system, central nervous system (such as cerebrospinal fluid), excretory system, digestive system, salivary’ system, skeletal system (such as synovial fluid), extracellular matrix, or a combination thereof.

[0332] Embodiment 28. The hybrid bioelectronic device of any one of Embodiments 1 to 27, wherein the hybrid bioelectronic device is configured to modulate a biological function with the body of the subject, wherein the modulation is electrical, mechanical, thermal, electro-chemical, chemical, optical, or a combination thereof.

[0333] Embodiment 29. The hybrid bioelectronic device of Embodiment 28, wherein the modulation of the biological function comprises (i) stimulation of parts of the central nervous system (CNS), the peripheral nervous system (PNS) such as the Dorsal Root Ganglion, the immune system, or the heart, (ii) promotion of wound healing or tissue regeneration, or (iii) treatment of tumors.

[0334] Embodiment 30. The hybrid bioelectronic device of Embodiment 28 or 29, wherein the modulation of the biological function comprises excitatory and / or inhibitory neuromodulation. Embodiment 31. The hybrid bioelectronic device of any one of Embodiments 1 to 30, wherein the hybrid bioelectronic device is configured to provide sensing, computing, analysis, and / or feedback functionalities.

[0335] Embodiment 32. The hybrid bioelectronic device of any one of Embodiments 1 to 31, wherein the hybrid bioelectronic device is configured to wirelessly receive and / or transmit commands and / or data, and / or is capable of brain computer interfacing.

[0336] Embodiment 33. A method of making a hybrid bioelectronic device, the method comprising: manufacturing, via top-down microfabrication techniques or bottom-up chemical synthesis, a wirelessly actuatable component configured to interact with an externally applied stimulus, wherein the wirelessly actuatable component is substrate-free; and attaching a biological or chemical component to the wirelessly actuatable component to produce the hybrid bioelectronic device, wherein the hybrid bioelectronic device is configured to be administered into a body of a subject and thereafter recognize, via the biological or chemical component, a target region within the body and self-implant at the target region.

[0337] Embodiment 34. The method of Embodiment 32, wherein the manufacturing comprises: (a) depositing a sacrificial layer on a substrate, the sacrificial layer being removable by an etchant selected from tetramethylammonium hydroxide (TMAH) solution, XeF2, or other suitable chemical etchants; (b) forming sequential device layers on the sacrificial layer using deposition, growth, or spin-coating methods; (c) patterning the device lay ers using photolithographic techniques and optionally dry etching (including reactive ion etching or ionbeam milling) to define a lateral dimension ranging from about 5 nm to about 5 mm; (d) removing the sacrificial layer to release the wirelessly actuatable component from the substrate, making them substrate-free; and (e) collecting the released wirelessly actuatable component using a harvesting process comprising filtration, centrifugation, or a combination thereof.

[0338] Embodiment 35. A method comprising: introducing one or more hybrid bioelectronic devices (such as those of any one of Embodiments 1 to 32) into a body of a subject in need thereof; permitting the one or more hybrid bioelectronic devices to travel within the body of the subject to a target region and to self-implant at the target region; and applying a stimulus to a wirelessly actuatable component of the self-implanted one or more hybrid bioelectronic devices which are configured to interact with the stimulus.

[0339] Embodiment 36. The method of Embodiment 35, wherein the one or more hybrid bioelectronic devices comprise a plurality of the hybrid bioelectronic device of any one of Embodiments 1 to 32, optionally wherein the plurality consists of (i) a single type of hybrid bioelectronic device, or (ii) a combination of two or more different types of hybrid bioelectronic devices.

[0340] Embodiment 37. The method of Embodiment 36, wherein the different types of hybrid bioelectronic devices can independently transduce at least two different types of externally applied stimulus into energy.

[0341] Embodiment 38. The method of any one of Embodiments 35 to 37. wherein the introducing of the one or more hybrid bioelectronic devices into the body of the subject is done via a non-invasive or minimally -invasive route of administration, optionally as part of a pharmaceutically acceptable fluid composition.

[0342] Embodiment 39. The method of any one of Embodiments 35 to 38. wherein the one or more of the hybrid bioelectronic device travel within the body of the subject via a fluid system within the subject, and optionally cross a biological barrier, such as the blood brain barrier, CSF- brain barrier, CSF-spinal cord barrier or CSF-DRG barrier.

[0343] Embodiment 40. The method of Embodiment 39, wherein the fluid system is a vascular system (such as blood or plasma), lymph system, central nervous system (such as cerebrospinal fluid), excretory system, digestive system, salivary system, skeletal system (such as synovial fluid), extracellular matrix, or combination thereof.

[0344] Embodiment 41. The method of any one of Embodiments 35 to 40, w herein: the target region is within the patient’s brain, spine, spinal cord, Dorsal Root Ganglion, peripheral nervous system, heart, lungs, or kidneys; and / or the target region comprises a region of inflammation.

[0345] Embodiment 42. The method of any one of Embodiments 35 to 41 , wherein the subject is in need of treatment for Alzheimer’s disease, multiple sclerosis, ischemic stroke, brain tumor, chronic pain, spinal cord injury', peripheral nerve injury, paralysis, essential tremor, a movement disorder, cardiovascular disease, autoimmune disease, arthritis, an infection, or a mental disorder, such as depression / anxiety, schizophrenia, post-traumatic stress disorder, autism, bipolar disorder, and disruptive mood dysregulation disorder.

[0346] Embodiment 43. The method of any one of Embodiments 35 to 42, w herein the applied stimulus is an externally applied electrical, optical, electromagnetic, magnetic or acoustic stimulus.

[0347] Embodiment 44. The method of any one of Embodiments 35 to 43, wherein the wirelessly actuatable component comprises a transducer which converts the externally applied stimulus into electrical, mechanical, thermal or electro-chemical energy'.

[0348] Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of skill in the art to which the disclosed invention belongs. Publications cited herein and the materials for which they are cited are specifically incorporated by reference.

[0349] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention. Such equivalents are intended to be encompassed by the following claims.

Claims

CLAIMSThat which is claimed is:

1. A hybrid bioelectronic device comprising: a wirelessly actuatable component configured to interact with an externally applied stimulus; and a biological or chemical component attached to the wirelessly actuatable component, wherein the hybrid bioelectronic device is configured to be administered into a body of a subject and thereafter recognize, via the biological or chemical component, a target region within the body and self-implant at the target region.

2. The hybrid bioelectronic device of claim 1, wherein the wirelessly actuatable component has a size that is between 5 nm and 5 mm.

3. The hybrid bioelectronic device of claim 1, wherein the wirelessly actuatable component comprises a transducer configured to convert the externally applied stimulus into electrical, mechanical, thermal, or electro-chemical energy.

4. The hybrid bioelectronic device of claim 3, wherein the wirelessly actuatable component further comprises (i) a nanoelectronic circuit configured to be powered by the transducer, and / or (ii) an antenna for wireless communication.

5. The hybrid bioelectronic device of claim 3, wherein the transducer is optical, magnetostrictive, piezoelectric, or magnetoelectric.

6. The hybrid bioelectronic device of claim 1, wherein the externally applied stimulus is an electrical, optical, electromagnetic, magnetic, or acoustic stimulus, or a combination thereof.

7. The hybrid bioelectronic device of claim 5, wherein the transducer is a planar structure or a non-planar structure, optionally wherein the non-planar structure is spherical or irregular in shape.

8. The hybrid bioelectronic device of claim 5, wherein the transducer is an optical transducer which comprises a substrate-free optoelectronic device.

9. The hybrid bioelectronic device of claim 8, wherein the substrate-free optoelectronic device consists of an active layer and optionally one or more additional layers selected from a group consisting of an anode layer; a cathode layer; an electron transport layer (ETL); a hole transport layer (HTL); a layer for improving interface properties or stability; a protective layer; and combinations thereof.

10. The hybrid bioelectronic device of claim 9, wherein the active layer comprises an organic semiconductor.

11. The hybrid bioelectronic device of claim 9, wherein the active layer is formed of at least one donor material, at least one acceptor material, or a combination thereof.

12. The hybrid bioelectronic device of claim 11. wherein the at least one donor material is an organic polymer, optionally wherein the organic polymer comprises poly[(2,6-(4,8-bis(5- (2-ethylhexyl-3-fluoro)thiophen-2-yl)-benzo[l,2-b:4,5-b']dithiophene))-alt-(5,5-(T,3'-di- 2-thienyl-5',7'-bis(2-ethylhexyl)benzo[r,2'-c:4',5'-c']dithiophene-4, 8-dione)] (PM6), poly[(2,6-(4,8-bis(5-(2-ethylhexyl-3-fluoro)thiophen-2-yl)-benzo[l,2-b:4,5- b']dithiophene))-alt-(5,5-(r,3'-di-2-thienyl-5',7'-bis(2-ethylhexyl)benzo[T,2'-c:4'.5'- c']dithiophene-4.8-dione)] (DI 8), poly[(4,4'-bis(2-ethylhexyl)dithieno[3,2-b:2'.3'- d]silole)-2,6-diyl-alt-(2,l,3-benzothiadiazole)-4,7-diyl] (PSBTBT), poly[N-9'- heptadecanyl-2,7-carbazole-alt-5,5'-(4',7'-di-2-thienyl-2',T,3'-benzothiadi azole)] (PCDTBT), poly(3-hexylthiophene) (P3HT), poly(2,6-(4,4-bis-(2-ethylhexyl)-4H- cyclopenta[2.1-b:3,4-b']dithiophene)-alt-4,7-(2,l,3-benzothiadiazole) (PCPDTBT), poly(2,5-thiophene) (P3T), poly(3,4-ethylenedioxythiophene) (PEDOT), poly(9,9- dioctylfluorene) (PFO), poly(2,5-bis(3-alkylthiophen-2-yl)thieno[3,2-b]thiophene) (PBTTT), poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS), poly(3,6-dioctylthieno[3.2-b]thiophene) (P3OT), poly(2,5-thiophene vinylene) (PTV), polythiophenes with various alkyl side chains and substitutions. poly(3.4- difluorothiophene-co-bithiophene) (PFTB), poly(2,6-carbazole) (PCz), poly(9,9- dioctylfluorene-co-bithiophene) (F8T2), poly (3-hexylthiophene) (P3HT), poly[2,6-(4,4- bis-(2-ethylhexyl)-4H-cyclopenta[2,l-b;3,4-b']dithiophene)-alt-4,7(2,l,3- benzothiadiazole)] (PCPDTBT). or a combination thereof.

13. The hybrid bioelectronic device of claim 11. wherein the at least one acceptor material is an organic polymer, optionally wherein the organic polymer comprises L8-BO (5,5'-Bis((2-butyloctyl)oxy)-[l,r:3',l"-terphenyl]-4,4"-dicarbaldehyde), Y6 (2.2'-((2Z,2'Z)- ((12,13-bis(2-ethylhexyl)-3,9-diundecyl-12,13-dihydro-[l,2,5]thiadiazolo[3,4- e]thieno[2,3":4',5']thieno[2',3,:4,5]pyrrolo[3,2-g]thieno[2,,3,:4,5]thieno[3,2-b]indole- 2,10-diyl)bis(methanylylidene))bis(5,6-difluoro-3-oxo-2,3-dihydro-lH-indene-2,l- diylidene))dimalononitrile), Z8 (2,2'-((2Z,2'Z)-((12,13-bis(2-ethylhexyl)-3,9-diundecyl- 12,13-dihydro-[l,2,5]thiadiazolo[3,4-e]thieno[2,3":4',5']thieno[2'.3':4,5]pyrrolo[3,2- g]thieno[2,.3':4,5]thieno[3.2-b]indole-2,10-diyl)bis(methanylylidene))bis(3-oxo-2,3- dihydro-lH-indene-5,6-dicarbonitrile))dimalononitrile), o-BTP-eC9 (2,2'-((2Z,2'Z)- ((12,13-bis(2-ethylhexyl)-3,9-bis(2-(2-ethoxyethoxy)ethyl)-12,13-dihydro- [l,2,5]thiadiazolo[3,4-e]thieno[2,3":4',5']thieno[2',3':4,5]pyrrolo[3,2- g]thieno[2'.3':4,5]thieno[3.2-b]indole-2,10-diyl)bis(methanylylidene))bis(3-oxo-2,3- dihydro-lH-indene-2,l-diylidene))dimalononitrile), P-2BTh-F (3,9-bis(2-methylene((3- (l,l-dicyanomethylene)-6,7-difluoro)indanone))-5,5,l l,l l-tetrakis(4- hexylphenyl)dithieno[2,3-d:2',3'-d']sindaceno[l,2-b:5,6-b']dithiophene), and PC70BM ([6,6]-Phenyl-C71-butyric acid methyl ester, PCBM (Phenyl-C61-butyric acid methyl ester), ITIC (Indacenodithieno|3,2-b]thiophene-2.8-dicarboximide), IT-M (Indacenodithiophene-based molecule), EH-IDTBR (2,2’-((2Z,2’Z)-((12,13-bis(2- ethylhexyl)-3,9-diundecyl-12,13-dihydro-[l,2,5]thiadiazolo[3,4-e]pyrazine-2,2,6,6- tetrayl))bis(4,5-dihydrothieno[3,4-b][l,4]dioxine-5,7-diyl))bis(methanylylidene))bis(3- ethyl-2-thioxothiazolidin-4-one), [6.6] -phenyl-C61 -butyric acid methyl ester (PCBM), or a combination thereof.

14. The hybrid bioelectronic device of claim 9, wherein the anode and cathode layers are electrically conducting materials.

15. The hybrid bioelectronic device of claim 9, wherein the ETL layer comprises (i) an organic polymer, optionally wherein the organic polymer comprises poly[[2,7-bis(2-ethylhexyl)1,2, 3,6,7, 8-hexahydro-l, 3,6, 8-tetraoxobenzo[lmn]phenanthroline-4, 9-diyl] - 2.5-thiophenediyl[9,9-bis[3'-((N.N-dimethyl)-N-ethylamino)propyl]- 9H-fluorene-2.7-diyl]-2,5-thiophenediyl] (PNDIT-F3N); (ii) a metal or metal oxide, optionally wherein the metal oxide comprises zinc oxide (ZnO), titanium dioxide (TiCh); or (iii) a combination thereof.1 . The hybrid bioelectronic device of claim 9, wherein the HTL layer comprises (i) an organic polymer, optionally wherein the organic polymer comprises poly(3,4- ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS); (ii) a metal or metal oxide, optionally wherein the metal oxide comprises molybdenum trioxide (MoCh), tungsten trioxide (WO3), vanadium pentoxide (V2O5); or (iii) a combination thereof.

17. The hybrid bioelectronic device of claim 9, wherein the layers for improving interface properties comprise: a) a metal, metal alloy, or metal compound; b) a PEDOT:PSS or a variant thereof; c) a carbon-based material; or d) a combination thereof.

18. The hybrid bioelectronic device of claim 5, wherein the transducer is an optical transducer which is based on inorganic materials.

19. The hybrid bioelectronic device of claim 5, wherein the transducer is an optical transducer and the optical transducer comprises one or more inorganic semiconductorbased devices, optionally wherein the one or more inorganic semiconductor-based devices comprise silicon nanowires with p-i-n junctions, III-V semiconductor nanowires, heterostructure nanowires, GaAs-based thin film planar devices, crystalline silicon-based thin film planar devices, amorphous silicon-based thin film planar devices, CIGS-based thin film planar devices, or CdTe-based thin film devices, optionally wherein the optical transducer comprises one or more encapsulating or passivating layers.

20. The hybrid bioelectronic device of claim 5, wherein the transducer is an optical transducer and the optical transducer is based on porphyrin-based materials.

21. The hybrid bioelectronic device of claim 5, wherein the transducer is a magnetoelectric transducer and the magnetoelectric transducer is formed either as a composite of magnetostrictive and piezoelectric materials or as a single-phase material exhibiting intrinsic coupling between magnetic and electric properties.

22. The hybrid bioelectronic device of claim 21. wherein the magnetoelectric transducer is formed as a composite of magnetostrictive and piezoelectric materials and the composite is either in a core-shell structure or in a planar layered structure.

23. The hybrid bioelectronic device of claim 1, wherein the biological agent comprises at least one of biological cells, viruses, cell membranes, extracellular vesicles, synthetics cells, or biomolecules, optionally wherein the biological agent is genetically engineered or biologically or chemically modified.

24. The hybrid bioelectronic devices of claim 23, wherein the biological cells comprises immune cells, stem / progenitor cells, glial cells, structural cells, or blood cells, or combinations thereof.

25. The hybrid bioelectronic device of claim 1, wherein the biological or chemical agent is attached to the wirelessly actuatable component by a linker that is permanent, selfdegrading, or cleavable by a biological signal or by exposure to an external stimulus.

26. The hybrid bioelectronic device of claim 1, wherein the hybrid bioelectronic device is configured to be able to be transported through a fluid system of the body of the subject.

27. The hybrid bioelectronic device of claim 25, wherein the fluid system comprises a vascular system, lymph system, central nervous system, excretory system, digestive system, salivary system, skeletal system, extracellular matrix, or a combination thereof.

28. The hybrid bioelectronic device of claim 1, wherein the hybrid bioelectronic device is configured to modulate a biological function with the body of the subject, wherein the modulation is electrical, mechanical, thermal, electro-chemical, chemical, optical, or a combination thereof.

29. The hybrid bioelectronic device of claim 28. wherein the modulation of the biological function comprises (i) stimulation of parts of the central nervous system (CNS), the peripheral nervous system (PNS), the immune system, or the heart, (ii) promotion of wound healing or tissue regeneration, or (iii) treatment of tumors.

30. The hybrid bioelectronic device of claim 28, wherein the modulation of the biological function comprises excitatory and / or inhibitory neuromodulation.

31. The hybrid bioelectronic device of claim 1, wherein the hybrid bioelectronic device is configured to provide sensing, computing, analysis, and / or feedback functionalities.

32. The hybrid bioelectronic device of claim 1, wherein the hybrid bioelectronic device is configured to wirelessly receive and / or transmit commands and / or data, and / or is capable of brain-computer interfacing.

33. A method of making a hybrid bioelectronic device, the method comprising: manufacturing, via top-down microfabrication techniques or bottom-up chemical synthesis, a wirelessly actuatable component configured to interact with an externally applied stimulus, wherein the wirelessly actuatable component is substrate-free; and attaching a biological or chemical component to the wirelessly actuatable component to produce the hybrid bioelectronic device, wherein the hybrid bioelectronic device is configured to be administered into a body of a subject and thereafter recognize, via the biological or chemical component, a target region within the body and self-implant at the target region.

34. The method of claim 33, wherein the manufacturing comprises:(a) depositing a sacrificial layer on a substrate, the sacrificial layer being removable by an etchant:(b) forming sequential device layers on the sacrificial layer using deposition, growth, or spin-coating methods;(c) patterning the device layers using photolithographic techniques and optionally dry etching to define a lateral dimension ranging from about 5 nm to about 5 mm;(d) removing the sacrificial layer to release the wirelessly actuatable component from the substrate, making them substrate-free; and(e) collecting the released wirelessly actuatable component using a harvesting process comprising filtration, centrifugation, or a combination thereof.

35. A method comprising: introducing one or more hybrid bioelectronic devices into a body of a subject in need thereof; permitting the one or more hybrid bioelectronic devices to travel within the body of the subject to a target region and to self-implant at the target region; andapplying a stimulus to a wirelessly actuatable component of the self-implanted one or more hybrid bioelectronic devices which are configured to interact with the stimulus.

36. The method of claim 35, wherein the one or more hybrid bioelectronic devices comprise a plurality of the hybrid bioelectronic device of any one of claims 1 to 32, optionally wherein the plurality consists of (i) a single type of hybrid bioelectronic device, or (ii) a combination of two or more different types of hybrid bioelectronic devices.

37. The method of claim 36, wherein the different types of hybrid bioelectronic devices can independently transduce at least two different types of externally applied stimulus into energy.

38. The method of claim 35, wherein the introducing of the one or more hybrid bioelectronic devices into the body of the subject is done via a non-invasive or minimally-invasive route of administration, optionally as part of a pharmaceutically acceptable fluid composition.

39. The method of claim 37, wherein the one or more of the hybrid bioelectronic device travel within the body of the subject via a fluid system within the subject, and optionally cross a biological barrier.

40. The method of claim 38, wherein the fluid system is a vascular system, lymph system, central nervous system, excretory system, digestive system, salivary system, skeletal system, extracellular matrix, or combination thereof.

41. The method of claim 35, wherein: the target region is within the patient’s brain, spine, spinal cord, Dorsal Root Ganglion, peripheral nervous system, heart, lungs, or kidneys; and / or the target region comprises a region of inflammation.

42. The method of claim 35, wherein the subject is in need of treatment for Alzheimer’s disease, multiple sclerosis, ischemic stroke, brain tumor, chronic pain, spinal cord injury, peripheral nerve injury, paralysis, essential tremor, a movement disorder, cardiovascular disease, autoimmune disease, arthritis, an infection, or a mental disorder, such asdepression / anxiety. schizophrenia, post traumatic stress disorder, autism, bipolar disorder, and disruptive mood dysregulation disorder.

43. The method of claim 35, wherein the applied stimulus is an externally applied electrical, optical, electromagnetic, magnetic or acoustic stimulus.

44. The method of claim 42, wherein the wirelessly actuatable component comprises a transducer which converts the externally applied stimulus into electrical, mechanical, thermal or electro-chemical energy.

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