Amnion-sourced engineered electric patch wound dressing as well as preparation method and application thereof

Abstract

The invention provides an amnion-sourced engineered electric patch wound dressing as well as a preparation method and application thereof, and belongs to the technical field of medical wound dressings. The engineered electric patch wound dressing is of a double-layer structure and comprises a moisture absorption layer and a volatilization layer, the double-layer structure is formed through physical pressing, the moisture absorption layer is a natural amnion, and the volatilization layer is hydrogel doped with carbon black particles. According to the engineered electric patch wound dressing, the biological activity of the natural amnion is utilized, and the wound repairing speed is increased; hydrogel doped with carbon black particles is constructed, a volatilization layer with the high moisture volatilization characteristic is prepared, meanwhile, by means of the moisture absorption characteristic of a natural amnion, wound exudate is efficiently absorbed, the engineered electric patch wound dressing with the wet electricity characteristic is obtained, and under an electric field constructed by the patch, the wound dressing can be used for wound dressing with the wet electricity characteristic. The repair degree of wound tissues is remarkably improved, meanwhile, differentiation of nerve cells is greatly promoted, and high-level repair of wounds is facilitated.

Description

Technical Field

[0001] The invention belongs to the technical field of medical wound dressings, and in particular relates to an engineered electric patch wound dressing derived from amniotic membrane, and a preparation method and application thereof. Background Art

[0002] The wound healing process mainly includes the hemostasis phase, the inflammatory phase, the proliferative phase and the maturation phase. Among them, wound inflammation, especially persistent chronic inflammation, greatly disrupts the balance of the wound microenvironment, and the levels of inflammatory factors, reactive oxygen species and proteases are highly upregulated, resulting in a large infiltration of immune cells, obstruction of angiogenesis in the wound, and severe damage to wound re-epithelialization, which poses a major obstacle to the wound healing process. Although traditional medical gauze dressings have the advantages of low price and easy application, they cannot significantly promote the healing of wounds, and gauze has a poor barrier effect on infection sources such as bacteria and fungi, and there is still a large risk of wound infection. Therefore, the development of efficient wound healing dressings has important clinical practice value and social significance.

[0003] In recent years, with the continuous development of regenerative medicine and tissue engineering technology, a variety of new treatment methods including growth factor therapy, gene therapy, stem cell therapy, and electrical stimulation therapy have achieved positive therapeutic effects in the field of wound repair. Based on the above-mentioned new treatment methods, a variety of new dressings, such as film dressings, sponge dressings, hydrogel dressings and other emerging carriers have come out one after another, bringing a wealth of wound repair product options to patients. However, existing wound dressings still have the challenges of insufficient wound healing efficiency and weak anti-inflammatory effects. In addition, vascular regeneration and nerve repair during wound remodeling also face major obstacles. Therefore, it is urgent to invent a new dressing that can efficiently promote wound healing and reshape the wound microenvironment at a high level. Summary of the invention

[0004] In view of this, the purpose of the present invention is to provide an engineered electric patch wound dressing derived from amniotic membrane, and a preparation method and application thereof. The present invention achieves a high level of wound repair by combining the biological benefits of amniotic membrane with the physical effects of electrical stimulation.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The invention provides an engineered electric patch wound dressing derived from amniotic membrane. The electric patch wound dressing comprises a hygroscopic layer and a volatile layer. The hygroscopic layer is based on natural amniotic membrane, and the volatile layer is prepared from a hydrogel doped with carbon black particles.

[0007] Preferably, the natural amniotic membrane is derived from any one of sheep, human and bovine amniotic membrane, and the thickness of the hygroscopic layer is 10 to 200 μm.

[0008] Preferably, the thickness of the volatile layer is 0.5 to 5 mm, and the particle size of the carbon black particles in the volatile layer is in the range of 10 to 1000 nm.

[0009] Preferably, the hydrogel is one or more of polyvinyl alcohol, polyacrylic acid, polymethacrylic acid, polyacrylamide, chitosan, alginate, hyaluronic acid, cellulose and gelatin.

[0010] Preferably, the mass ratio of the carbon black particles to the hydrogel is 1:10-200.

[0011] The present invention provides a method for preparing the engineered electrical patch wound dressing, comprising the following steps:

[0012] 1) Incubate the amniotic membrane in a complete medium containing vitamins to obtain a hygroscopic layer;

[0013] 2) mixing the carbon black particles and the hydrogel, and performing a cross-linking reaction to obtain a volatile layer;

[0014] 3) Pressing the hygroscopic layer and the volatile layer together to obtain an engineered electrical patch wound dressing.

[0015] Preferably, the complete culture medium comprises vitamin C, vitamin D2 and vitamin D3, wherein the concentration of vitamin C is 0.01 to 10 mM, the concentration of vitamin D2 is 0.01 to 10 mM, and the concentration of vitamin D3 is 0.01 to 10 mM.

[0016] Preferably, the cross-linking reaction time is 25 to 35 minutes.

[0017] Preferably, the pressing pressure is 0.4-0.7 MPa, and the pressing time is 25-35 min.

[0018] The present invention also provides the use of the engineered electric patch wound dressing in preparing a product for promoting wound repair.

[0019] Compared with the prior art, the present invention has the following beneficial effects: the present invention provides an engineered electric patch wound dressing derived from amniotic membrane, and the engineered electric patch wound dressing has a double-layer structure, which is a natural amniotic membrane hygroscopic layer and a volatile layer doped with carbon black particle hydrogel. Under the principle of hydrovoltaic power generation, the engineered electric patch wound dressing can effectively realize the absorption and evaporation of water vapor at the wound site, form a water gradient difference inside, thereby driving the directional movement of ions, and forming a continuous and stable physical electrical stimulation at the wound site. In addition, under the biological benefits of natural amniotic membrane such as anti-inflammatory and pro-proliferation, the engineered electric patch wound dressing constructed by the present invention exhibits a high level of wound healing and remodeling efficiency. In the present invention, natural amniotic membrane is used as the substrate, which has good tissue repair ability, can effectively inhibit the inflammatory response at the wound site, promote cell proliferation and migration at the wound site, thereby significantly accelerating the wound healing rate, and at the same time, the hygroscopic properties of natural amniotic membrane are utilized, combined with the principle of hydrovoltaic power generation, and a volatile layer with a high water evaporation efficiency is further developed, thereby constructing an amniotic membrane-derived engineered electric patch wound dressing with a double-layer structure. The engineered electric patch wound dressing can form a stable physical electrical stimulation under the stimulation of moisture and humidity at the wound position, which can not only promote cell proliferation and migration, but also promote angiogenesis and induce mesenchymal stem cells to differentiate into nerve cells. Therefore, the present invention is not only to accelerate the closed healing of the wound, but also to promote the effective remodeling of the wound environment, which is an innovation in the field of wound dressings. And the engineered electric patch wound dressing of amniotic origin provided by the present invention has a simple structure and composition, a simple preparation process, and good biocompatibility. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is a scanning electron microscope analysis photograph of the engineered electrical patch (ElectroSAM) derived from sheep amnion prepared in Example 1;

[0021] Figure 2 This is a graph showing the mechanical properties test results of the engineered electroSAM derived from sheep amniotic membrane prepared in Example 1;

[0022] Figure 3 This is a graph showing the electrical performance test results of the engineered electroSAM derived from sheep amniotic membrane prepared in Example 1;

[0023] Figure 4 This is a diagram showing the evaluation results of the anti-inflammatory effect of the engineered electroSAM derived from sheep amniotic membrane prepared in Example 1;

[0024] Figure 5 This is a diagram showing the evaluation results of the migration-promoting ability of the engineered electroSAM derived from sheep amnion prepared in Example 1;

[0025] Figure 6 This is a diagram showing the evaluation results of the angiogenesis-promoting ability of the engineered electrostrip (ElectroSAM) derived from sheep amniotic membrane prepared in Example 1;

[0026] Figure 7 This is a diagram showing the evaluation results of the ability of the engineered electrostrip (ElectroSAM) derived from sheep amniotic membrane prepared in Example 1 to promote neural differentiation;

[0027] Figure 8 This is a graph showing the evaluation results of the wound repair ability of the engineered electrostrip (ElectroSAM) derived from sheep amniotic membrane prepared in Example 1 in diabetic mice. DETAILED DESCRIPTION

[0028] The invention provides an engineered electric patch wound dressing derived from amniotic membrane. The electric patch wound dressing comprises a hygroscopic layer and a volatile layer. The hygroscopic layer is based on natural amniotic membrane, and the volatile layer is prepared from a hydrogel doped with carbon black particles.

[0029] In the present invention, the natural amniotic membrane is derived from any one of sheep, human and bovine amniotic membrane, preferably sheep-derived amniotic membrane; the thickness of the hygroscopic layer is 10-200 μm, preferably 50-150 μm, and more preferably 70-100 μm; the thickness of the volatile layer is 0.5-5 mm, preferably 0.6-2 mm, and more preferably 0.7-1 mm; the particle size range of the carbon black particles in the volatile layer is 10-1000 nm, preferably 100-700 nm, and more preferably 300-500 nm; the mass ratio of the carbon black particles to the hydrogel is 1:10-200, preferably 1:50-150, and more preferably 1:100; the hydrogel is one or more of polyvinyl alcohol, polyacrylic acid, polymethacrylic acid, polyacrylamide, chitosan, alginate, hyaluronic acid, cellulose, and gelatin.

[0030] The present invention provides a method for preparing the engineered electrical patch wound dressing, comprising the following steps:

[0031] 1) Incubate the amniotic membrane in a complete medium containing vitamins to obtain a hygroscopic layer;

[0032] 2) mixing carbon black particles and hydrogel, performing a cross-linking reaction, and obtaining a volatile layer;

[0033] 3) Pressing the hygroscopic layer and the volatile layer together to obtain an engineered electrical patch wound dressing.

[0034] In the present invention, the amniotic membrane is placed in a complete medium containing vitamins and incubated to obtain a hygroscopic layer. The method for obtaining the amniotic membrane is as follows: the sheep amniotic membrane (SAM) is separated and extracted from the sheep placenta under sterile conditions, and washed with sterile phosphate buffered saline (PBS) containing antibiotics for 2 to 8 times, preferably 3 to 6 times, and more preferably 5 times. The antibiotics are penicillin, streptomycin, neomycin and amphotericin B, wherein the content of penicillin is 1 to 50 μg / mL, preferably 10 to 40 μg / mL, and more preferably 30 μg / mL, the content of streptomycin is 1 to 50 μg / mL, preferably 10 to 40 μg / mL, and more preferably 30 μg / mL, the content of neomycin is 1 to 50 μg / mL, preferably 10 to 40 μg / mL, and more preferably 20 μg / mL, and the content of amphotericin B is 1 to 50 μg / mL, preferably 1 to 10 μg / mL, and more preferably 3 μg / mL. Subsequently, the amniotic membrane is immersed in a mixed solution of DMEM culture medium and glycerol and stored at -80°C for standby use. The volume ratio of the DMEM culture medium and glycerol is 1:0.5-2, preferably 1:0.7-1.5, and more preferably 1:1. Before use, the preserved amniotic membrane is immersed in a DMEM culture medium containing 10% FBS (w / v) and vitamin C, vitamin D2, and vitamin D3 for incubation to enhance the activity of the cells contained in the amniotic membrane, thereby obtaining enhanced biological functions of the amniotic membrane. The concentration of the vitamin C is 0.01-10mM, preferably 0.1-5mM, and more preferably 1mM, the concentration of the vitamin D2 is 0.01-10mM, preferably 0.1-5mM, and more preferably 1mM, and the concentration of the vitamin D3 is 0.01-10mM, preferably 0.1-5mM, and more preferably 1mM.

[0035] In the present invention, carbon black particles are mixed with hydrogel, and a cross-linking reaction is performed to obtain a volatile layer. The cross-linking reaction is to mix the carbon black particles with the hydrogel, add glutaraldehyde, vortex for 2 to 8 minutes, preferably 4 to 6 minutes, and more preferably 5 minutes, and stand for cross-linking reaction; the mass ratio of glutaraldehyde to hydrogel is 1:10 to 30, preferably 1:15 to 25, and more preferably 1:20, and the cross-linking reaction time is 25 to 35 minutes, preferably 27 to 33 minutes, and more preferably 30 minutes.

[0036] In the present invention, the hygroscopic layer and the volatile layer are pressed together to obtain an engineered electrical patch wound dressing. The pressing is to press the hygroscopic layer obtained in step 1) and the volatile layer obtained in step 2) by physical compression, the pressing pressure is 0.4-0.7MPa, preferably 0.45-0.6MPa, more preferably 0.5MPa, and the pressing time is 25-35min, preferably 27-33min, more preferably 30min.

[0037] The present invention also provides the use of the engineered electric patch wound dressing in preparing a product for promoting wound repair.

[0038] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0039] Example 1

[0040] The preparation method of the engineered electrostrip patch (ElectroSAM) derived from sheep amniotic membrane is as follows:

[0041] (1) Sheep amniotic membrane (SAM) was isolated and extracted from sheep placenta under sterile conditions and washed five times with sterile phosphate buffered saline (PBS) containing antibiotics (30 μg / mL penicillin, 30 μg / mL streptomycin, 20 μg / mL neomycin and 3 μg / mL amphotericin B). Subsequently, the amniotic membrane was immersed in a mixed solution of DMEM culture medium and glycerol (1:1, v / v) and stored at -80°C. Before use, the preserved amniotic membrane was immersed in DMEM culture medium containing 10% FBS (w / v) and vitamin C (1 mM), vitamin D2 (1 mM), and vitamin D3 (1 mM) and incubated for 12 h to enhance the activity of the cells contained in the amniotic membrane, thereby obtaining enhanced biological functions of the amniotic membrane.

[0042] (2) The carbon black particles were mixed into a polyvinyl alcohol (PVA) aqueous solution (10%, w / w) (carbon black particles / PVA, 1:100, w / w), stirred evenly, and then cross-linked with glutaraldehyde (glutaraldehyde / PVA, 1:20, w / w), and reacted at 25°C for 30 minutes to obtain a hydrogel volatile layer doped with carbon black particles.

[0043] (3) The prepared SAM and the hydrogel volatile layer doped with carbon black particles were physically compressed for 30 min under a pressure of 0.5 MPa to press the SAM and the volatile layer together to prepare an engineered electrical patch (ElectroSAM) derived from the amniotic membrane of sheep.

[0044] like Figure 1Shown are scanning electron microscope (SEM) analysis images of the bottom layer, top layer and cross section of ElectroSAM.

[0045] Example 2

[0046] The preparation method of the engineered electrostrip patch (ElectroSAM) derived from sheep amniotic membrane is as follows:

[0047] (1) Sheep amniotic membrane (SAM) was isolated and extracted from sheep placenta under sterile conditions and washed three times with sterile phosphate buffered saline (PBS) containing antibiotics (30 μg / mL penicillin, 30 μg / mL streptomycin, 20 μg / mL neomycin and 3 μg / mL amphotericin B). Subsequently, the amniotic membrane was immersed in a mixed solution of DMEM culture medium and glycerol (1:0.5, v / v) and stored at -80°C. Before use, the preserved amniotic membrane was immersed in DMEM culture medium containing 10% FBS (w / v) and vitamin C (0.1 mM), vitamin D2 (0.1 mM), and vitamin D3 (0.1 mM) and incubated for 12 h to enhance the activity of the cells contained in the amniotic membrane, thereby obtaining enhanced biological functions of the amniotic membrane.

[0048] (2) The carbon black particles were mixed into a polyvinyl alcohol (PVA) aqueous solution (10%, w / w) (carbon black particles / PVA, 1:50, w / w), stirred evenly, and then cross-linked with glutaraldehyde (glutaraldehyde / PVA, 1:10, w / w), and reacted at 25°C for 25 minutes to obtain a hydrogel volatile layer doped with carbon black particles.

[0049] (3) The prepared SAM and the hydrogel volatile layer doped with carbon black particles were physically compressed for 35 minutes under a pressure of 0.4 MPa to press the SAM and the volatile layer together to prepare an engineered electrical patch (ElectroSAM) derived from the amniotic membrane of sheep.

[0050] Example 3

[0051] The preparation method of the engineered electrostrip patch (ElectroSAM) derived from sheep amniotic membrane is as follows:

[0052] (1) Sheep amniotic membrane (SAM) was isolated and extracted from sheep placenta under sterile conditions and washed 8 times with sterile phosphate buffered saline (PBS) containing antibiotics (30 μg / mL penicillin, 30 μg / mL streptomycin, 20 μg / mL neomycin and 3 μg / mL amphotericin B). Subsequently, the amniotic membrane was immersed in a mixed solution of DMEM culture medium and glycerol (1:2, v / v) and stored at -80°C. Before use, the preserved amniotic membrane was immersed in DMEM culture medium containing 10% FBS (w / v) and vitamin C (10 mM), vitamin D2 (10 mM), and vitamin D3 (10 mM) and incubated for 12 h to enhance the activity of the cells contained in the amniotic membrane, thereby obtaining enhanced biological functions of the amniotic membrane.

[0053] (2) The carbon black particles were mixed into a polyvinyl alcohol (PVA) aqueous solution (10%, w / w) (carbon black particles / PVA, 1:200, w / w), stirred evenly, and then cross-linked with glutaraldehyde (glutaraldehyde / PVA, 1:30, w / w), and reacted at 25°C for 35 minutes to obtain a hydrogel volatile layer doped with carbon black particles.

[0054] (3) The prepared SAM and the hydrogel volatile layer doped with carbon black particles were physically compressed for 25 minutes under a pressure of 0.7 MPa to press the SAM and the volatile layer together to prepare an engineered electrical patch (ElectroSAM) derived from the amniotic membrane of sheep.

[0055] Effect Example 1 Mechanical Properties Test of Engineered ElectroSAM Derived from Amniotic Membrane The engineered electroSAM derived from amniotic membrane (ElectroSAM) prepared in Example 1 and the mechanical tensile properties of SAM were tested using an electronic universal testing machine. Each sample was fixed on the universal testing machine, the tensile speed was set to 5 mm / min, and the stress-strain curve was drawn to examine the mechanical properties (including elastic modulus and ultimate tensile strength) of each material. The results are shown in FIG. Figure 2 As shown, the elastic modulus and ultimate tensile strength of natural amniotic membrane (SA) are 2.9 MPa and 0.9 MPa, respectively, and the elastic modulus and ultimate tensile strength of the engineered electrical patch (ElectroSAM) derived from amniotic membrane are 7.0 MPa and 1.6 MPa, respectively, indicating that the mechanical properties of ElectroSAM are significantly enhanced.

[0056] Effect Example 2 Electrical Performance Test of Engineered ElectroSAM Derived from Sheep Amniotic Membrane

[0057] The engineered electrostrip ElectroSAM prepared in Example 1 was placed in different humidity environments (RH=50%, 70%, 90%, 99%), and a digital multimeter was used to measure and record the voltage output of ElectroSAM within one hour. The output voltage-time curve is shown in FIG. Figure 3 As shown. The output voltage value of ElectroSAM is positively correlated with the relative humidity, and can stably form an output voltage of about 170mV under a relative humidity of 99%. It shows that the engineered electric patch ElectroSAM derived from the sheep amniotic membrane prepared in Example 1 can effectively realize the water vapor absorption and evaporation at the wound site, form a water gradient difference inside, thereby driving the directional movement of ions and forming a continuous and stable physical electrical stimulation at the wound site.

[0058] Effect Example 3 Evaluation of the anti-inflammatory effect of engineered electrostrips derived from sheep amniotic membrane (ElectroSAM)

[0059] Mouse mononuclear macrophages (RAW 264.7 cells, purchased from the Cell Resource Center of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) were cultured in DMEM complete medium containing 10% FBS at a rate of 2×10 per well. 5 The cells were seeded in a 24-well plate at a density of 100 cells / mL and incubated at 37°C for 24 hours. Lipopolysaccharide (LPS, 500 ng / mL) was used to stimulate the cells, and the engineered electric patch ElectroSAM derived from the sheep amnion prepared in Example 1 was used to incubate the LPS-stimulated cells for 24 hours. Subsequently, the cells were washed three times with PBS and incubated at 37°C for 20 minutes with serum-free DMEM medium containing 10 μmol / L DCFH-DA (ROS fluorescent probe) and 10 μg / mL Hoechst 33342 (nuclear dye). Subsequently, the cells were analyzed by fluorescence imaging using an inverted fluorescence microscope. The results are shown in Figure 2. Figure 4 As shown, incubation of ElectroSAM significantly reduced the ROS expression of macrophages under LPS treatment, indicating that ElectroSAM effectively inhibits inflammation.

[0060] Effect Example 4 Evaluation of the Migration-Promoting Ability of the Engineered ElectroSAM Derived from Sheep Amniotic Membrane

[0061] Mouse fibroblasts (NIH 3T3 cells, purchased from the Cell Resource Center of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) were cultured at a rate of 2×10 5The cells were inoculated in a 24-well plate at a density of 100 cells and grown at 37°C until 90% confluence. Subsequently, a 200 μL pipette tip was used to make a straight scratch in the well plate to construct a straight cell scratch. Subsequently, the complete medium containing 1.0 mM hydrogen peroxide (H2O2) was replaced and incubated with the engineered electrostrip ElectroSAM derived from the sheep amnion prepared in Example 1 (24 hours), and images were taken at predetermined time points (0h, 24h). The results are shown in FIG. Figure 5 As shown, in the control group under inflammatory environment, the 24h migration rate of cells was only 7.8%, while ElectroSAM treatment significantly promoted the migration rate of NIH 3T3 cells under H2O2 inflammatory environment, and the 24h migration rate reached 59.4%.

[0062] Effect Example 5 Evaluation of the angiogenesis-promoting ability of engineered electrostrips (ElectroSAM) derived from sheep amniotic membrane

[0063] Corning Matrigel (purchased from Corning, USA, Cat. No. 354234) was used to coat 48-well cell culture plates (100 μL / well) and incubated at 37°C for 30 minutes. Human umbilical vein endothelial cells (HUVEC cells, purchased from the Cell Resource Center of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) were seeded into Matrigel-coated 48-well plates (5×10 cells / well) using DMEM complete medium containing 10% FBS. 4 The cells were incubated with the engineered electrostripping patch ElectroSAM prepared in Example 1 for 6 hours, and the photos were analyzed by microscope. Figure 6 As shown, ElectroSAM greatly promoted angiogenesis.

[0064] Effect Example 6 Evaluation of the ability of engineered electrostrips (ElectroSAM) derived from sheep amniotic membrane to promote neural differentiation

[0065] Mouse bone marrow-derived mesenchymal stem cells (MSCs, primary self-extracted) were seeded into confocal culture dishes (2×10 per dish) using DMEM complete medium containing 10% FBS. 4 cells), and incubated with the engineered electrostrip ElectroSAM derived from the sheep amnion prepared in Example 1 for 10 days. Subsequently, the cells were washed three times with PBS, fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 for 10 minutes, and blocked with 1% bovine serum albumin (BSA) for 30 minutes. Next, the above cells were incubated with Nestin antibody and β3-Tubulin antibody at 4°C overnight. Subsequently, FITC-labeled fluorescent secondary antibody was used for incubation at 37°C for 1 hour, and fluorescence imaging analysis was performed using a confocal microscope (CLSM) after DAPI staining. The results are shown in Figure 7As shown, ElectroSAM treatment greatly enhanced the expression of neuronal markers Nestin and β3-Tubulin.

[0066] Effect Example 7 Evaluation of the ability of engineered electrostrips (ElectroSAM) derived from sheep amniotic membrane to promote wound repair in diabetic mice

[0067] C57BL / 6 mice (male, purchased from Nanjing Qinglongshan Co., Ltd.) were used and fed with a high-fat diet for 12 weeks. Mice with blood sugar levels higher than 11.1 mmol / L were randomly selected to obtain a male diabetic mouse model. On this basis, a full-thickness skin wound model was constructed, and the mice were anesthetized using isoflurane through a gas anesthesia machine. The skin on the back of the mice was removed using a razor and a depilatory cream. After disinfection with alcohol and iodine tincture, a wound with a diameter of 5 mm was cut on the back. The engineered electric patch ElectroSAM derived from the amniotic membrane prepared in Example 1 was used as a wound dressing. Fresh dressings were replaced every other day, and the wound healing rate was calculated on days 0, 5, 10, and 15. Specifically, the wound healing process of the relevant groups is as follows. Figure 8 As shown. ElectroSAM significantly accelerated the wound healing process in diabetic mice.

[0068] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. An engineered electrical patch wound dressing derived from amniotic membrane, characterized in that: The electric patch wound dressing comprises a hygroscopic layer and a volatile layer, wherein the hygroscopic layer is based on natural amniotic membrane, and the volatile layer is prepared from a hydrogel doped with carbon black particles.

2. The engineered electrical patch wound dressing according to claim 1, characterized in that: The natural amniotic membrane is derived from any one of sheep, human and bovine amniotic membrane, and the thickness of the hygroscopic layer is 10 to 200 μm.

3. The engineered electrical patch wound dressing according to claim 1, characterized in that: The thickness of the volatile layer is 0.5-5 mm, and the particle size of the carbon black particles in the volatile layer is in the range of 10-1000 nm.

4. The engineered electrical patch wound dressing according to claim 1, characterized in that: The hydrogel is one or more of polyvinyl alcohol, polyacrylic acid, polymethacrylic acid, polyacrylamide, chitosan, alginic acid, hyaluronic acid, cellulose and gelatin.

5. The engineered electrical patch wound dressing according to claim 3 or 4, characterized in that: The mass ratio of the carbon black particles to the hydrogel is 1:10-200.

6. The method for preparing the engineered electrical patch wound dressing according to any one of claims 1 to 5, characterized in that: The steps include: 1) Incubate the amniotic membrane in a complete medium containing vitamins to obtain a hygroscopic layer; 2) mixing carbon black particles and hydrogel, and performing a cross-linking reaction to obtain a volatile layer; 3) Pressing the hygroscopic layer and the volatile layer together to obtain an engineered electrical patch wound dressing.

7. The preparation method according to claim 6, characterized in that: The complete culture medium comprises vitamin C, vitamin D2 and vitamin D3, wherein the concentration of vitamin C is 0.01 to 10 mM, the concentration of vitamin D2 is 0.01 to 10 mM, and the concentration of vitamin D3 is 0.01 to 10 mM.

8. The preparation method according to claim 6, characterized in that: The cross-linking reaction time is 25 to 35 minutes.

9. The preparation method according to claim 6, characterized in that: The pressing pressure is 0.4-0.7 MPa, and the pressing time is 25-35 min.

10. Use of the engineered electrical patch wound dressing according to any one of claims 1 to 5 or the engineered electrical patch wound dressing obtained according to the preparation method according to any one of claims 6 to 9 in the preparation of a product for promoting wound repair.

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