Conductive hydrogel-mediated mesenchymal stem cell electrical stimulation induced differentiation device

Abstract

The invention provides a conductive hydrogel-mediated mesenchymal stem cell electrical stimulation induced differentiation device, and belongs to the technical field of biomedical engineering and medical instruments, and the device comprises a programmable body surface activation electrical stimulation unit which adopts near-field coupling energy supply and communication, adopts body surface attachment without a lead, and outputs symmetrical biphase pulses; the conductive hydrogel interface unit takes a light / enzyme / ion cross-linked biocompatible polymer as a skeleton, is doped with a conductive filler, is injected to a nerve injury section through a needle head, is cured into a coating / sleeve-shaped structure in situ at the level of 10 seconds to minute through light, conforms to nerve geometry and maintains low-impedance fitting; the MSC is suspended in conductive hydrogel precursor fluid, and the MSC is derived from umbilical cord / bone marrow / fat; the pain points of unstable stimulating dose, device-cell separation, MSC function attenuation and the like in the prior art are solved, and a standardized and high-curative-effect solution is provided for peripheral nerve injury repair.

Description

Technical Field

[0001] This invention relates to the fields of biomedical engineering and medical device technology, specifically to the interdisciplinary fields of tissue engineering and regenerative medicine, and bioelectronics / neuroelectric stimulation, and particularly to a conductive hydrogel-mediated mesenchymal stem cell electrical stimulation-induced differentiation device. Background Technology

[0002] Peripheral nerve injury (PNI) can lead to long-term sensory and motor dysfunction. Traditional repair strategies (direct suturing, autologous nerve transplantation, and nerve conduits) still have unstable long-term functional recovery in scenarios with long defects, mixed nerves, and delayed repair. The core challenge lies in the difficulty of simultaneously achieving rapid axonal regeneration, myelin sheath reconstruction, and optimization of the immune microenvironment.

[0003] In existing technologies, low-frequency pulsed electrical stimulation (ES) has been shown to promote nerve regeneration by upregulating pathways such as cAMP / PKA and BDNF / TrkB, but it has significant limitations: transcutaneous / surface stimulation (TENS / microcurrent) is affected by skin impedance, and the effective dose in the damaged segment is uncontrollable; wired rigid implanted electrodes (such as cuff electrodes) are precise but are prone to mechanical mismatch, infection, and secondary damage; wireless biodegradable stimulators avoid the problem of wires, but they are mostly single-modal interventions of the device on the tissue and are not synergistic with cell therapy.

[0004] Conductive hydrogels (such as PEDOT:PSS and GelMA-MXene systems) combine soft-wet matching with low impedance characteristics and have been used in neural interfaces. However, most existing products are passive conductive bridges and are not deeply coupled with programmable electrical stimulation sources or MSC carriers. MSCs are widely used for PNI repair due to their low immunogenicity and strong paracrine ability, and can be induced into Schwann-like cells (SCLC). However, traditional transplantation has problems such as low survival rate, unstable phenotype, and suppressed paracrine function.

[0005] Recent studies have shown that electrical stimulation can activate MSCs to differentiate into SCLCs and enhance paracrine function. However, current protocols are still in a loosely coupled state of electrical stimulation and materials, and materials and cells. They lack an integrated design that incorporates programmable electrical stimulation, low-impedance conductive interfaces, in situ differentiation of MSCs, and paracrine amplification. This results in unstable stimulation doses, inefficient energy-cell conversion, and MSC function decline, making it difficult to meet clinical repair needs.

[0006] Therefore, this invention proposes a conductive hydrogel-mediated mesenchymal stem cell electrical stimulation-induced differentiation device. Summary of the Invention

[0007] This invention provides a conductive hydrogel-mediated mesenchymal stem cell electrical stimulation-induced differentiation device to solve the aforementioned technical problems.

[0008] This invention provides a conductive hydrogel-mediated mesenchymal stem cell electrical stimulation-induced differentiation device, comprising: Programmable surface-activated electrical stimulation unit: It uses near-field coupling for power supply and communication, is attached to the body surface without wires, and outputs symmetrical biphasic pulses; Conductive hydrogel interface unit: Based on a biocompatible polymer that can be cross-linked by light, enzymes, and ions, doped with 5-60 wt% conductive filler, with conductivity of... It possesses shear-thinning rheological properties. When injected into the nerve injury segment through a 27~30G needle, it is solidified in situ in ten seconds to minutes by 405 / 365nm light into a 0.1~1.5mm thick covering / sleeve-like structure that conforms to the nerve geometry and maintains low impedance adhesion. In-situ carriers of bioactive MSCs: MSCs are suspended in a conductive hydrogel precursor solution, with an encapsulation concentration of (1~10)×10⁻⁶. 6 MSCs / mL, injection volume 5~20μL per damaged segment, MSC sources include umbilical cord / bone marrow / fat; The conductive hydrogel interface unit serves as both an MSC carrier and a soft electrode, stably transmitting the pulses output by the electrical stimulation unit to the MSCs, inducing the MSCs to differentiate into Schwann cells and enhancing neurotrophic and immune-regulating paracrine functions.

[0009] Preferably, in the conductive hydrogel interface unit, the mass ratio of conductive filler to polymer backbone is 15-40wt%; the polymer backbone is GelMA, combined with 0.05-0.5wt% photoinitiator LAP, and cured by irradiation with a 405nm light source for 10-60s. After curing, the elastic modulus is 1-50kPa, which is mechanically matched with peripheral nerve tissue.

[0010] Preferably, the pulse parameters of the programmable body surface activation electrical stimulation unit are set as follows: frequency 1~50Hz, pulse width 50~500μs / phase, tissue field strength 20~300mV / mm, daily stimulation 30~120min, and total treatment course 1~21 days.

[0011] Preferably, Schwann cells, peripheral nerve progenitor cells or exosomes are used in combination, or MSCs are pretreated with hypoxia / factor pre-excitation.

[0012] Preferably, the conductive hydrogel interface unit is further provided with a conductive gradient, microgroove / fiber orientation structure, or segmented layered design to induce MSC / axon directional growth and electric field focusing.

[0013] Preferred options also include: A closed-loop monitoring module integrating impedance, temperature, and strain microsensors communicates with the body surface activation electrical stimulation unit for closed-loop adjustment of electrical stimulation dosage and recording of treatment data.

[0014] Preferably, the flexible energy-receiving components of the conductive hydrogel interface unit and the programmable body surface activation electrical stimulation unit are made of biodegradable materials.

[0015] Preferred options also include: The accompanying kit contains a sterile conductive hydrogel precursor solution, an MSC suspension storage container, a disposable microneedle tube, a body surface activation electrical stimulation unit, and a curing light source, and all components are sterilized.

[0016] Compared with the prior art, the beneficial effects of this application are: Stimulation dose is stable and controllable: solving the problem of transdermal stimulation dose drift; Significantly enhanced MSC function: The synergistic effect of electrical stimulation and conductive microenvironment increased the differentiation rate of MSCs into SCLC by 2.3 times, the secretion of BDNF reached 168.13 pg / mL (2.71 times higher than the unstimulated group), and the levels of NGF and IL-10 increased by 1.75 times each. Excellent in vivo repair efficacy: In a rat sciatic nerve injury model, 8 weeks after surgery, the SFI was 31.88% higher than that of the injury control group, the gastrocnemius muscle preservation rate reached 95.38% (close to the sham surgery group), the CMAP amplitude increased by 470.1%, and the myelin sheath thickness reached 1.18μm (close to the physiological value of 1.26μm). Safety and surgical procedure optimization: The biodegradable design avoids secondary surgery, and needle injection and second-level curing shorten the intraoperative operation time to within 5 minutes; Mechanism is explainable and standardizable: Single-cell transcriptomics confirmed the enrichment of the MAPK / BDNF pathway, and the parameters were programmable to achieve cross-individual standardization, providing a basis for multi-center clinical application.

[0017] Other features and advantages of the invention will be set forth in the following description, and will be apparent in part from the description, or may be learned by practicing the invention. The objects and other advantages of the invention may be realized and obtained by means of the structures particularly pointed out in the written description and the accompanying drawings.

[0018] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description

[0019] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used in conjunction with embodiments of the invention to explain the invention and do not constitute a limitation thereof. In the drawings: Figure 1 This is a schematic diagram of the overall principle of a conductive hydrogel-mediated mesenchymal stem cell electrical stimulation-induced differentiation device in an embodiment of the present invention; Figure 2 This is a characterization diagram of the conductive hydrogel properties in an embodiment of the present invention; Figure 3 This is a diagram illustrating the in vitro induced differentiation of MSCs in an embodiment of the present invention; Figure 4 This is a diagram of single-cell transcriptome analysis in an embodiment of the present invention; Figure 5 The chart shows the in vivo repair effect in rats in this embodiment of the invention. Detailed Implementation

[0020] The preferred embodiments of the present invention will be described below with reference to the accompanying drawings. It should be understood that the preferred embodiments described herein are for illustration and explanation only and are not intended to limit the present invention.

[0021] This invention provides a conductive hydrogel-mediated mesenchymal stem cell electrical stimulation-induced differentiation device, comprising: Programmable surface-activated electrical stimulation unit: It uses near-field coupling for power supply and communication, is attached to the body surface without wires, and outputs symmetrical biphasic pulses; Conductive hydrogel interface unit: Based on a biocompatible polymer that can be cross-linked by light, enzymes, and ions, doped with 5-60 wt% conductive filler, with conductivity of... It possesses shear-thinning rheological properties. When injected into the nerve injury segment through a 27~30G needle, it is solidified in situ in ten seconds to minutes by 405nm light into a 0.1~1.5mm thick covering / sleeve-like structure that conforms to the nerve geometry and maintains low impedance adhesion. In-situ carriers of bioactive MSCs: MSCs are suspended in a conductive hydrogel precursor solution, with an encapsulation concentration of (1~10)×10⁻⁶. 6 MSCs / mL, injection volume 5~20μL per damaged segment, MSC sources include umbilical cord / bone marrow / fat; The conductive hydrogel interface unit serves as both an MSC carrier and a soft electrode, stably transmitting the pulses output by the electrical stimulation unit to the MSCs, inducing the MSCs to differentiate into Schwann cells and enhancing neurotrophic and immune-regulating paracrine functions.

[0022] Preferably, in the conductive hydrogel interface unit, the mass ratio of conductive filler to polymer backbone is 15-40wt%; the polymer backbone is GelMA, combined with 0.05-0.5wt% photoinitiator LAP, and cured by irradiation with a 405nm light source for 10-60s. After curing, the elastic modulus is 1-50kPa, which is mechanically matched with peripheral nerve tissue.

[0023] Preferably, the pulse parameters of the programmable body surface activation electrical stimulation unit are set as follows: frequency 1~50Hz, pulse width 50~500μs / phase, tissue field strength 20~300mV / mm, daily stimulation 30~120min, and total treatment course 1~21 days.

[0024] Preferably, Schwann cells, peripheral nerve progenitor cells or exosomes are used in combination, or MSCs are pretreated with hypoxia / factor pre-excitation.

[0025] Preferably, the conductive hydrogel interface unit is further provided with a conductive gradient, microgroove / fiber orientation structure, or segmented layered design to induce MSC / axon directional growth and electric field focusing.

[0026] Preferred options also include: A closed-loop monitoring module integrating impedance, temperature, and strain microsensors communicates with the body surface activation electrical stimulation unit for closed-loop adjustment of electrical stimulation dosage and recording of treatment data.

[0027] Preferably, the flexible energy-receiving components of the conductive hydrogel interface unit and the programmable body surface activation electrical stimulation unit are made of biodegradable materials.

[0028] Preferred options also include: The accompanying kit contains a sterile conductive hydrogel precursor solution, an MSC suspension storage container, a disposable microneedle tube, a body surface activation electrical stimulation unit, and a curing light source, and all components are sterilized.

[0029] Example 1: Preparation and characterization of Mo / GelMA conductive hydrogel: Preparation: Dissolve GelMA powder (preferably 5–15% w / v), add LAP (preferably 0.05–0.5% w / v) and sterile Mo nanoparticles according to... Disperse evenly to obtain the precursor fluid.

[0030] Formation: The sciatic nerve injury segment is clinically injected (27–30G) into the surface of the sciatic nerve injury segment and cured in situ for ten seconds by irradiation with a 405 nm light source (preferably 0.2–1.0 mm thick).

[0031] Characterization: SEM showed that the Mo particles were uniformly dispersed and the pore structure was continuous; electrochemical impedance spectroscopy (EIS) showed that the low-frequency impedance was significantly lower than that of undoped GelMA; in vitro degradation showed a controllable trend and good early compatibility.

[0032] Function: To provide a low-impedance, flexible interface for stable ES conduction and cell anchoring.

[0033] Example 2: In situ electrical stimulation of MSCs induces differentiation and paracrine amplification (in vitro): Cells and Culture: Umbilical cord MSCs were embedded in the hydrogel of Example 1 and cultured for 24 h before ES was started; ES scheme: 20 Hz, 100 μs / phase, ~100 mV / mm, 60 min / day × 7 days; Key findings: Proliferation: The Gel / Mo+ES group showed a 4.9-fold increase in proliferation at 7 days compared to 1 day; at 3 days, the Gel / Mo+ES group had a proliferation rate of 19.18±0.55, which was higher than that of the Gel / Mo group (13.36±0.44) (P<0.05). Phenotype: Directed growth was observed, with consistent enhancement of S100β (MSC→SCLC trend). Secretion: Compared with the Gel / Mo group without ES, BDNF increased by 2.71 times (61.94→168.13 pg / mL, p<0.001), NGF increased by 1.75 times, and IL-10 increased by 1.75 times (both p<0.001). Neural effects: In Transwell co-culture, Gel / Mo+ES significantly increased neurite length to 81.57±14.96 μm (p<0.001); microelectrode array (MEA) showed a spontaneous firing frequency of +387.7% (p<0.001).

[0034] Conclusion: Conductive microenvironment + programmed ES in situ induces MSCs to SCLC phenotype and amplifies neurotrophic / immune paracrine secretion.

[0035] Example 3: Single-cell transcriptomics validation of electrophysiological reprogramming: Design: Comparing MSC and ES groups (34,809 cells in total), UMAP clustering showed significant separation; Pathways: ES enriched MAPK, axonal regulation, FAK / PTK2, etc. (FDR<0.05, NES≥1.5); Simulated timing / potential: scTour / CytoTRACE / Monocle2 showed that maturation and differentiation potential decreased along the simulated timeline; ES increased the proportion of MSC_Stromal_Remodeling (BDNF, COL1A1, MMPs) and decreased MSC_Cycling; integration with the damaged neural atlas suggested enhanced interaction with Schwann cells, with the BDNF axis as the core.

[0036] Parameters: 20 Hz, 100 μs / phase, ~100 mV / mm, 60 min / day × 7 days.

[0037] Conclusion: The molecular basis of electrophysiological reprogramming leading to paracrine network enhancement was verified at the single-cell scale.

[0038] Example 4: In vivo validation of a rat sciatic nerve injury model: Model and drug administration: Sciatic nerve clipping; MSC dose preferably 5 × 10 4 Cells / 10 μL / damaged segment, Gel / Mo needle injection for in-situ solidification and combined with ES treatment (same parameters as in Example 2).

[0039] Functional analysis: At 8 weeks, SFI was significantly better in the device group than in the control group (ΔSFI vs LC +31.88±3.63; vs MSC +27.75±3.23; vs ES +19.69±3.37; all p<0.001). Muscle preservation: The gastrocnemius muscle preservation rate was 95.38% in the device group (98.76% in Sham); compared with LC / MSC / ES, it was +53.21 / +28.20 / +18.61% respectively (p≤0.006). Electrophysiology: CMAP amplitude was +470.1% compared to LC, latency 16.7%; also better than MSC / ES (both p<0.001; latency p<0.001 / 0.042). Histology: Myelin thickness 1.18±0.04 μm (near Sham 1.26±0.07), S100β intensity and positive area +65.5% and +77.2%, respectively (vs LC, both p<0.001). Safety: Major organ pathology is normal; hydrogel and peripheral electrodes are biodegradable and do not require removal.

[0040] Conclusion: ES alone is effective but insufficient; significant gains can be obtained by synergizing with activatable MSCs.

[0041] like Figure 1 The diagram shows the overall principle of the device, which is an integrated coupled system consisting of a programmable surface activation electrical stimulation unit, a conductive hydrogel interface unit, and a bioactive unit that carries MSCs in situ. Electrical stimulation forms a stable equivalent electric field through (hydrogel → tissue), and MSCs are induced to transform into Schwann-like cells (SCLCs) in situ by electrical stimulation and enhance paracrine function, which is used for peripheral nerve repair.

[0042] like Figure 2 The figure shows the characterization of the conductive hydrogel, including SEM morphology, impedance spectrum, mechanical curves and degradation curves, which confirm its low impedance, mechanical matching and degradability; providing an interfacial basis for stable conduction of electrical stimulation and cell anchoring.

[0043] like Figure 3 The image shows the in vitro differentiation of MSCs. Immunofluorescence shows the distribution of S100β-positive cells, and the bar chart shows the secretion of BDNF / NGF / IL-10, confirming the enhancement of phenotypic differentiation and paracrine function.

[0044] like Figure 4 The diagram shown is a single-cell transcriptome analysis diagram, including UMAP clustering, pathway enrichment bubble diagram, and pseudo-temporal trajectory, revealing the molecular mechanism of electrical stimulation-induced MSC differentiation.

[0045] like Figure 5The chart shown illustrates the in vivo repair effect in rats, confirming the therapeutic efficacy of the device.

[0046] The device of this invention is applicable to peripheral nerve injury repair (suturing, clamping, short / medium defect catheter reconstruction, etc.), and can also be extended to scenarios that require in-situ activation of MSCs and regulation of the microenvironment, such as chronic peripheral nerve lesions, tendon / muscle nerve innervation reconstruction, and repair of peripheral spinal cord branches.

[0047] The conductive hydrogel-mediated electrostimulation-induced differentiation device for mesenchymal stem cells (MSCs) provided by this invention, compared with existing technologies such as percutaneous stimulation, rigid implanted electrodes, passive conductive hydrogels, or simple cell therapy, forms a continuous closed loop of programmable electrical stimulation → low-impedance conductive hydrogel interface → in-situ electrostimulation differentiation and paracrine amplification of MSCs. This directly solves key problems such as the difficulty in stabilizing the stimulation dose, device-cell separation, MSC functional decline, and complex surgical procedures, achieving the following technical effects: 1) Stable, focused electrical stimulation dose delivery: Structure-Mechanism Relationship: Injectable, in-situ photocurable Mo / GelMA conductive hydrogels form a flexible, low-modulus, low-impedance interface at the damaged site, effectively transmitting symmetrical biphasic pulses from surface wireless (e.g., NFC) outputs to the target area, reducing dose drift caused by skin / soft tissue impedance and geometric attenuation. Compared to percutaneous or rigid electrodes, the electric field distribution is more uniform, and the dose reached is more reproducible, providing a stable prerequisite for programmable activation at the cell scale. The entire injection-curing process is completed within the surgical field, achieving formation in seconds / tens of seconds, reducing positioning errors and intraoperative time. The supplementary injection operation parameters, photocuring conditions, and patch fixation methods are as follows: GelMA: 5–15% w / v; preferably 7.5–12%; more preferably 10% (methylation degree 50–80%); photoinitiator: preferably LAP (Lithiumphenyl-2,4,6-trimethylbenzoylphosphinate) 0.03–0.10% w / v; preferably 0.05%. Light source: Wavelength (λ): 405±10nm (visible light, better than UV in terms of cell compatibility); Irradiance (irradiance I): 5–50mW / cm²; preferably ≤30mW / cm² in vivo. Energy density (dose E): E=I×t (J / cm²). Commonly used dose window for cell compatibility: 1–10J / cm²; preferably 2–6J / cm².

[0048] 2) In situ electro-induced differentiation and enhanced paracrine function of MSCs: Electrophysiological reprogramming: Under optimized parameters (e.g., 20 Hz, 100 μs / phase, approximately 100 mV / mm, 60 min / day × 7 days) applied in a conductive microenvironment, MSCs showed directional orientation enhancement consistent with S100β, suggesting a transformation towards Schwann-like cell (SCLC) phenotype. Proliferation and activity: The Gel / Mo+ES group proliferated 4.9 times on day 7 compared to day 1; on day 3, the proliferation rate was 19.18 ± 0.55, significantly higher than that of the Gel / Mo group (13.36 ± 0.44) (P < 0.05), indicating that the physical-electrical stimulation microenvironment provided by the device simultaneously enhanced cell activity and quantity. Paracrine amplification: Compared with the non-ES group, BDNF increased by 2.71 times (61.94→168.13 pg / mL, p<0.001), and NGF and IL-10 each increased by 1.75 times (p<0.001); in contactless co-culture, neurite length increased to 81.57±14.96 μm (p<0.001), and MEA recorded spontaneous firing frequency +387.7% (p<0.001). This indicates that electrical stimulation is effectively converted into a quantitative enhancement of MSC phenotype and secretion via the hydrogel interface, and produces a clear neural support effect.

[0049] 3) Mechanism-level explainability and repeatability: Single-cell transcriptomic evidence: In a comparison of 34,809 cells, the ES group was significantly separated from the MSC group; ES cells were enriched in pathways such as MAPK, axonal regulation, and FAK / PTK2 (FDR < 0.05, NES ≥ 1.5); the proportion of MSC_Stromal_Remodeling (including BDNF, COL1A1, and MMPs) increased, while MSC_Cycling decreased; integration with the injury neural atlas suggested enhanced MSC-Schwan cell interaction. This molecular atlas directly explains the causal chain of electrical stimulation—conductive interface—cell reprogramming—paracrine enhancement, improving the reproducibility and extrapolation of the results.

[0050] 4) Significant improvement in in vivo multiscale efficacy: At 8 weeks of the rat sciatic nerve clipping model, the SFI in the device group was significantly improved compared with the control (ΔSFI vs LC +31.88±3.63; vs MSC +27.75±3.23; vs ES +19.69±3.37; all p<0.001); gait dragging was significantly reduced, and the gastrocnemius muscle preservation rate was 95.38%, close to the sham-operated group (98.76%), which was +53.21 / +28.20 / +18.61% compared with LC / MSC / ES (p≤0.006), indicating higher reinnervation efficiency. CMAP amplitude was +470.1% compared with LC, and latency was... 16.7%; significantly better than MSC / ES (amplitude p<0.001; latency p<0.001 / 0.042), demonstrating simultaneous improvement in conduction function and myelin remodeling. Near-physiological myelin bundle structure and myelin sheath thickness (1.18±0.04μm, close to Sham 1.26±0.07), S100β intensity and positive area +65.5% and +77.2%, respectively (vsLC, both p<0.001). Compared to the current situation where ES alone is effective but insufficient, and MSC alone is prone to decay, device-level coupling achieves significant synergistic gains.

[0051] 5) Favorable remodeling of the immune microenvironment: Cytokine profile: In the device group, a significant increase in the anti-inflammatory factor IL-10 was observed, accompanied by a remodeling of the inflammatory profile, suggesting the establishment and maintenance of an M2-like repair environment, consistent with structural repair and conduction restoration. This verifies that the present invention is not simply about conduction / stimulation, but rather achieves joint regulation of immunity, nutrition, and remyelination by inducing MSCs in situ.

[0052] 6) Long-term interface stability and biosafety: The conductive hydrogel maintains a coordinated stability of conductivity, mechanics, and geometry in vivo, reducing output drift and impedance rise; safety and biodegradability: major organ pathologies are normal; the hydrogel and peripheral electrodes are biodegradable and do not require removal, reducing the risk of secondary surgery and long-term foreign body reactions. It meets the safety requirement of natural withdrawal after clinical treatment.

[0053] 7) Simplified Procedure and Clinical Accessibility: Needle injection and rapid curing allow for implantation and interface construction completed within minutes; the non-guided thread control of the surface patch improves compliance. Frequency, pulse width, field strength, and treatment duration can all be digitally set and reused, facilitating parameter standardization and multi-center promotion for different individuals and injury types. It avoids donor site damage and cost associated with autologous nerve harvesting, reduces the need for secondary retrieval surgery and follow-up costs, and improves rehabilitation efficiency and quality of life.

[0054] In this embodiment, a method for implementing a conductive hydrogel-mediated mesenchymal stem cell electrical stimulation-induced differentiation device includes the following steps: Step 1: Preoperative preparation: Prepare Mo / GelMA precursor fluid (containing MSC suspension); set ES parameters (frequency, pulse width, field strength, treatment course).

[0055] Step 2: Delivery and curing: Under a microscope, inject the pre-existing body fluid around the damaged area, and expose it to external light to quickly cross-link and form a conductive coating; if necessary, add a flexible fixing ring to ensure adhesion.

[0056] Step 3: Surface activation: Align the skin patch with the implantation site and start the preset program; 20Hz, 100μs / phase, ~100mV / mm, 60min / day × 7 days is a preferred treatment course.

[0057] Step 4: Follow-up and Exit: The device gradually degrades in the body after the treatment course; if an optional sensor is used, impedance changes are recorded to assess interface stability.

[0058] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A conductive hydrogel-mediated device for inducing differentiation of mesenchymal stem cells through electrical stimulation, characterized in that, include: Programmable surface-activated electrical stimulation unit: It uses near-field coupling for power supply and communication, is attached to the body surface without wires, and outputs symmetrical biphasic pulses; Conductive hydrogel interface unit: Based on a biocompatible polymer that can be cross-linked by light, enzymes, and ions, doped with 5-60 wt% conductive filler, with conductivity of... It possesses shear-thinning rheological properties. When injected into the nerve injury segment through a 27~30G needle, it is solidified in situ in ten seconds to minutes by 405 / 365nm light into a 0.1~1.5mm thick covering / sleeve-like structure that conforms to the nerve geometry and maintains low impedance adhesion. In-situ carriers of bioactive MSCs: MSCs are suspended in a conductive hydrogel precursor solution, with an encapsulation concentration of (1~10)×10⁻⁶. 6 MSCs / mL, injection volume 5~20μL per damaged segment, MSC sources include umbilical cord / bone marrow / fat; The conductive hydrogel interface unit serves as both an MSC carrier and a soft electrode, stably transmitting the pulses output by the electrical stimulation unit to the MSCs, inducing the MSCs to differentiate into Schwann cells and enhancing neurotrophic and immune-regulating paracrine functions.

2. The conductive hydrogel-mediated mesenchymal stem cell electrical stimulation-induced differentiation device according to claim 1, characterized in that, In the conductive hydrogel interface unit, the mass ratio of conductive filler to polymer backbone is 15-40wt%; the polymer backbone is GelMA, combined with 0.05-0.5wt% photoinitiator LAP, and is cured by irradiation with a 405nm light source for 10-60s. After curing, the elastic modulus is 1-50kPa, which is mechanically matched with peripheral nerve tissue.

3. The conductive hydrogel-mediated mesenchymal stem cell electrical stimulation-induced differentiation device according to claim 1, characterized in that, The pulse parameters of the programmable body surface activation electrical stimulation unit are set as follows: frequency 1~50Hz, pulse width 50~500μs / phase, tissue field strength 20~300mV / mm, daily stimulation 30~120min, total treatment course 1~21 days.

4. The conductive hydrogel-mediated mesenchymal stem cell electrical stimulation-induced differentiation device according to claim 1, characterized in that, Combine Schwann cells, peripheral neural progenitor cells or exosomes, or pre-treat MSCs with hypoxia / factor pre-excitation.

5. The conductive hydrogel-mediated mesenchymal stem cell electrical stimulation-induced differentiation device according to claim 1, characterized in that, The conductive hydrogel interface unit is also provided with conductive gradient, microgroove / fiber orientation structure, or segmented layered design to induce MSC / axon directional growth and electric field focusing.

6. The conductive hydrogel-mediated mesenchymal stem cell electrical stimulation-induced differentiation device according to claim 1, characterized in that, Also includes: A closed-loop monitoring module integrating impedance, temperature, and strain microsensors communicates with the body surface activation electrical stimulation unit for closed-loop adjustment of electrical stimulation dosage and recording of treatment data.

7. The conductive hydrogel-mediated mesenchymal stem cell electrical stimulation-induced differentiation device according to claim 1, characterized in that, The flexible energy-receiving components of the conductive hydrogel interface unit and the programmable body surface activation electrical stimulation unit are made of biodegradable materials.

8. The conductive hydrogel-mediated mesenchymal stem cell electrical stimulation-induced differentiation device according to any one of claims 1-7, characterized in that, Also includes: The accompanying kit contains a sterile conductive hydrogel precursor solution, an MSC suspension storage container, a disposable microneedle tube, a body surface activation electrical stimulation unit, and a curing light source, and all components are sterilized.

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