An electro-bioactive synergistic efficient pro-angiogenic hydrogel material, its preparation method and use

Abstract

A highly efficient angiogenic gel material with synergistic electro-bioactivity, its preparation method, and its applications are disclosed. The preparation method involves cross-linking gel composed of gelatin methylpropionate hydrogel, piezoelectric nanoparticles, and mesoporous bioglass under the action of an aqueous photoinitiator. The mesoporous bioglass is prepared by emulsification of hexadecyltrimethylammonium bromide, tetraethyl orthosilicate, and a calcium source using a template method followed by high-temperature calcination. This invention offers the following advantages: its preparation process is simple, stable, highly reproducible, and biocompatible. When subjected to external mechanical forces, this gel material generates an electric field, eliminating the need for an external power source. It enhances the mesoporous bioglass via a wireless field to achieve a highly efficient angiogenic effect. The piezoelectric nanoparticles in this gel material not only provide the necessary electrical stimulation but also enhance the mechanical properties of the bulk hydrogel.

Description

Technical Field

[0001] This invention relates to the interdisciplinary field of biomedical materials and skin tissue engineering, and particularly to a highly efficient angiogenic gel material with synergistic electro-bioactivity, its preparation method, and its uses. Background Technology

[0002] The skin is the largest organ in the human body, composed of a three-tiered structure consisting of the epidermis, dermis, and subcutaneous tissue, all of which possess self-renewal and external protective capabilities. However, skin wounds caused by external factors such as surgery, stress, and burns, or pathological factors such as diabetes and vascular diseases, are very common. Wounds that penetrate deep into the subcutaneous tissue directly damage the local vascular network, leading to impaired nutrient and oxygen transport to the damaged tissue. This results in slow wound healing and a higher risk of complications such as infection, ulceration, and tissue necrosis, making it a core challenge in clinical skin wound repair.

[0003] Currently, hydrogels have become the preferred material for constructing skin wound repair materials due to their softness, water retention, biocompatibility, and flexible and adjustable functionality. However, hydrogel wound dressings developed in research generally lack good tissue regeneration properties. Especially when the skin is damaged to the subcutaneous tissue layer, the vascular network in that area is disrupted, preventing the delivery of nutrients to the damaged tissue, resulting in slow wound healing and subsequently triggering a series of complications.

[0004] Therefore, in order to address the shortcomings of existing technologies, it is essential to provide a highly efficient angiogenic gel material with synergistic electro-bioactivity, its preparation method, and its applications. Summary of the Invention

[0005] One objective of this invention is to overcome the shortcomings of existing technologies and provide a method for preparing a highly efficient angiogenic gel material with synergistic electro-bioactivity. This method for preparing the highly efficient angiogenic gel material with synergistic electro-bioactivity is simple, stable, and highly reproducible, making it suitable for large-scale industrial production.

[0006] The above-mentioned objectives of the present invention are achieved through the following technical measures: A method for preparing a highly efficient angiogenic gel material with synergistic electro-bioactivity is provided, which is prepared by cross-linking gel with gelatin methyl propionate hydrogel, piezoelectric nanoparticles and nanoporous bioglass under the action of an aqueous photoinitiator.

[0007] The nanoporous bioglass is prepared by emulsification of hexadecyltrimethylammonium bromide, tetraethyl orthosilicate and calcium source using a template method followed by high-temperature calcination.

[0008] Preferably, the above-mentioned nanoporous bioglass is prepared by the following steps: A1. Dissolve hexadecyltrimethylammonium bromide in water to obtain solution A1; A2. Add ethyl acetate to solution A1 to obtain solution A2; A3. Add the ammonia solution to solution A2 to obtain solution A3; A4. Add tetraethyl orthosilicate to solution A3 to obtain solution A4; A5. Add calcium nitrate tetrahydrate to solution A4 to react, and then a precipitate is obtained; A6. The precipitate is calcined to obtain the nanoporous bioglass.

[0009] The nanoporous bioglass, by weight, is prepared by the following steps: A1. Dissolve 1 to 4 parts of hexadecyltrimethylammonium bromide in 50 to 200 parts of water to obtain solution A1; A2. Add 20 to 60 parts of ethyl acetate to solution A1 to obtain solution A2; A3. Add 10 to 40 parts of 1 mol / L ammonia solution to solution A2 to obtain solution A3; A4. Add 5 to 25 parts of tetraethyl orthosilicate to solution A3 to obtain solution A4; A5. Add 2 to 20 parts of calcium nitrate tetrahydrate to solution A4 and react to obtain a precipitate; A6. The precipitate is calcined to obtain the nanoporous bioglass.

[0010] Preferably, the above-mentioned nanoporous bioglass is prepared by the following steps: A1. Add hexadecyltrimethylammonium bromide to water and control the temperature to 30℃~40℃, stir for 10min~15min to obtain solution A1; A2. Add ethyl acetate to solution A1 and stir for 30 min to 40 min to obtain solution A2; A3. Add the ammonia solution to solution A2 and stir for 15 min to 20 min to obtain solution A3; A4. Add tetraethyl orthosilicate to solution A3 and stir for 30 min to 40 min to obtain solution A4; A5. Add calcium nitrate tetrahydrate to solution A4 and react for 4 to 6 hours. Then collect the solid and wash it several times with deionized water and ethanol alternately. Then dry it at 50°C to 60°C for 12 to 18 hours to obtain the precipitate. A6. The precipitate is calcined at 600℃~900℃ and the sintering holding time is 2h~4h to obtain the nanoporous bioglass.

[0011] In the A5, the amount of deionized water used for each cleaning is 200 to 300 parts; the amount of ethanol used for each cleaning is 200 to 300 parts.

[0012] The procedure, by weight, is as follows: S1. Add 0.01 to 0.15 parts of aqueous photoinitiator to 15 to 30 parts of PBS solution to obtain solution I; S2. Add 0.5 to 10 parts of gelatin methyl propionate hydrogel to solution I to obtain solution II; S3. Add 0.2 to 5 parts of piezoelectric nanoparticles to solution II, and then treat with ultrasound to obtain solution III; S4. Add 0.6 to 15 parts of nanoporous bioglass to solution III, and then treat with ultrasound to obtain solution IV; S5. Solution IV is placed in a mold and irradiated with an ultraviolet light source to obtain a highly efficient angiogenic gel material with synergistic electro-biological activity.

[0013] The method for preparing the highly efficient angiogenic gel material with synergistic electro-bioactivity of the present invention comprises the following steps: S1. Add 0.01 to 0.15 parts of aqueous photoinitiator to 15 to 30 parts of PBS solution and stir at 40°C to 50°C for 5 to 20 minutes to obtain solution I; S2. Add 0.5 to 10 parts of gelatin methyl propionate hydrogel to solution I and stir at 60℃ to 70℃ for 5 min to 20 min to obtain solution II; S3. Add 0.2 to 5 parts of piezoelectric nanoparticles to solution II, and then sonicate for 10 to 30 minutes to obtain solution III, with an ultrasonic intensity of 0.3 W / cm. 2 ~0.6W / cm 2 ; S4. Add 0.6 to 15 parts of nanoporous bioglass to solution III, then sonicate for 10 to 30 minutes to obtain solution IV, with an ultrasonic intensity of 0.3 W / cm. 2 ~0.6W / cm 2 ; S5. Place solution IV into a mold and irradiate it with an ultraviolet light source for 100s to 300s to obtain a highly efficient angiogenic gel material with synergistic electro-biological activity.

[0014] Preferably, the calcium source mentioned above is calcium nitrate tetrahydrate.

[0015] Preferably, the above-mentioned gelatin methyl propionate hydrogel is prepared from methacrylic anhydride-modified gelatin.

[0016] Preferably, the piezoelectric nanoparticles are one or more of barium titanate, potassium sodium niobate, bismuth tungstate, and sodium bismuth titanate, and are prepared by hydrothermal reaction with tetrabutyl titanate and barium hydroxide octahydrate.

[0017] A second objective of this invention is to overcome the shortcomings of existing technologies and provide a highly efficient angiogenesis-promoting gel material with synergistic electro-bioactivity. This highly efficient angiogenesis-promoting gel material with synergistic electro-bioactivity can generate an electric field and has superior mechanical properties and good biocompatibility.

[0018] The above-mentioned objectives of the present invention are achieved through the following technical measures: A highly efficient angiogenic gel material with synergistic electro-bioactivity is provided, which is prepared by the above-mentioned method for preparing highly efficient angiogenic gel material with synergistic electro-bioactivity.

[0019] A third objective of this invention is to overcome the shortcomings of existing technologies by providing a highly efficient angiogenesis-promoting gel material with synergistic electro-bioactivity in the preparation of biomedical materials for skin wound repair. This highly efficient angiogenesis-promoting gel material, through the synergistic effect of radio fields and bioactivity, can generate an electric field to achieve highly efficient angiogenesis, making it particularly suitable for repairing deep subcutaneous tissue skin wounds with damaged vascular networks.

[0020] The above-mentioned objectives of the present invention are achieved through the following technical measures: This invention provides the use of a highly efficient angiogenesis-promoting gel material with synergistic electro-bioactivity in the preparation of biomedical materials for skin wound repair. The highly efficient angiogenesis-promoting gel material with synergistic electro-bioactivity is prepared by the aforementioned method for preparing highly efficient angiogenesis-promoting gel materials with synergistic electro-bioactivity.

[0021] This invention discloses a highly efficient angiogenesis-promoting gel material with synergistic electro-bioactivity, its preparation method, and its uses. The preparation method involves cross-linking gel composed of gelatin methylpropionate hydrogel, piezoelectric nanoparticles, and mesoporous bioglass under the action of an aqueous photoinitiator. The mesoporous bioglass is prepared by emulsifying hexadecyltrimethylammonium bromide, tetraethyl orthosilicate, and a calcium source using a template method followed by high-temperature calcination. This invention offers the following advantages: 1. The preparation process of the highly efficient angiogenesis-promoting gel material with synergistic electro-bioactivity is simple, stable, highly reproducible, and biocompatible. 2. This highly efficient angiogenesis-promoting gel material can generate an electric field when subjected to external mechanical force, eliminating the need for an external power source and enhancing the mesoporous bioglass through a wireless field to achieve a highly efficient angiogenesis-promoting effect. 3. The piezoelectric nanoparticles in this highly efficient angiogenesis-promoting gel material not only provide the necessary electrical stimulation but also enhance the mechanical properties of the bulk hydrogel. Detailed Implementation

[0022] The technical solution of the present invention will be further described with reference to the following embodiments. Unless otherwise specified, the experimental methods in the following embodiments are conventional methods. Unless otherwise specified, the raw materials and reagents used in the following embodiments can be purchased from conventional biochemical reagent stores or pharmaceutical companies. Attached Figure Description

[0023] The invention will be further described with reference to the accompanying drawings, but the contents of the drawings do not constitute any limitation on the invention.

[0024] Figure 1 The image shows the microstructure and elemental analysis of the nanoporous bioglass prepared in Example 4.

[0025] Figure 2 The image shows the microstructure and elemental analysis of the nanoporous bioglass prepared in Example 5.

[0026] Figure 3 The image shows the microstructure of the piezoelectric nanoparticles prepared in Example 5.

[0027] Figure 4 The images show the macroscopic morphology of the electro-bioactive synergistic and highly efficient vascular-promoting gel materials prepared in Examples 4 and 5, where 4A is the gel material prepared in Example 4 and 4B is the gel material prepared in Example 5.

[0028] Figure 5 The image shows the microstructure of the electro-bioactive synergistic and highly efficient vascular-promoting gel material prepared in Example 4.

[0029] Figure 6The image shows the microstructure of the electro-bioactive synergistic and highly efficient vascular-promoting gel material prepared in Example 5.

[0030] Figure 7 Fluorescence analysis of the electro-bioactive synergistic and highly efficient angiogenesis-promoting gel material in the blank group, comparative example 1, and example 5.

[0031] Figure 8 The positive control, comparative example 1, and example 5 are shown in the diagrams illustrating the synergistic and efficient hemolytic effect of the electro-bioactive vascular gel material on promoting blood vessel dissolution.

[0032] Example 1

[0033] A method for preparing a highly efficient vascular-promoting gel material with synergistic electro-bioactivity involves cross-linking gelatin methyl propionate hydrogel, piezoelectric nanoparticles, and nanoporous bioglass under the action of an aqueous photoinitiator.

[0034] The nanoporous bioglass was prepared by emulsification of hexadecyltrimethylammonium bromide, tetraethyl orthosilicate, and a calcium source via a template method followed by high-temperature calcination. The calcium source was calcium nitrate tetrahydrate. The nanoporous bioglass exhibits excellent pro-angiogenic bioactivity, and its ion release rate can be modulated under an electric field, achieving a synergistic effect with its electroactivity.

[0035] Gelatin methylpropionate hydrogel is prepared from methacrylic anhydride-modified gelatin. In this invention, the gelatin methylpropionate hydrogel is prepared using the method described in Chinese Patent Publication No. CN109251277 B, "A Potassium Sodium Niobate Nanoparticle Composite Hydrogel and Its Preparation Method and Application." Due to its three-dimensional network structure, the gelatin methylpropionate hydrogel has advantages such as good adhesion to the wound surface, isolation of bacteria and harmful particles, excellent moisture absorption, good biocompatibility, and non-adhesion to the wound.

[0036] The piezoelectric nanoparticles are one or more of barium titanate, potassium sodium niobate, bismuth tungstate, and sodium bismuth titanate, prepared by a hydrothermal reaction with tetrabutyl titanate and barium hydroxide octahydrate. In this invention, the piezoelectric nanoparticles are prepared using the method described in Chinese Patent Publication No. CN113209290B, "A Bismuth / Barium Titanate Heterojunction for Enhancing Acoustodynamic Antitumor Effects and Its Preparation Method," which describes the preparation method of barium titanate piezoelectric nanoparticles. The piezoelectric nanoparticles can generate a radio field under external mechanical force, while simultaneously enhancing the mechanical properties of the gel material.

[0037] It should be noted that this invention utilizes the piezoelectric properties of piezoelectric nanoparticles combined with the excellent angiogenesis-promoting properties of mesoporous bioglass to obtain a highly efficient angiogenesis-promoting gel material with synergistic electro-bioactivity. Furthermore, the electroactive hydrogel material can generate an electric field, and the mesoporous bioglass, in conjunction with this electric field, enhances its angiogenesis-promoting performance. The electroactive gel material of this invention does not require external electrodes; instead, it relies on the piezoelectric properties of the piezoelectric nanoparticles to generate an electric field through external mechanical force, stimulating surrounding cells via a wireless electric field.

[0038] The method for preparing the highly efficient angiogenic gel material with synergistic electro-bioactivity of the present invention comprises the following steps, in parts by weight: S1. Add 0.01 to 0.15 parts of aqueous photoinitiator to 15 to 30 parts of PBS solution to obtain solution I; S2. Add 0.5 to 10 parts of gelatin methyl propionate hydrogel to solution I to obtain solution II; S3. Add 0.2 to 5 parts of piezoelectric nanoparticles to solution II, and then treat with ultrasound to obtain solution III; S4. Add 0.6 to 15 parts of nanoporous bioglass to solution III, and then treat with ultrasound to obtain solution IV; S5. Solution IV is placed in a mold and irradiated with an ultraviolet light source to obtain a highly efficient angiogenic gel material with synergistic electro-biological activity.

[0039] Furthermore, the preparation method of the highly efficient angiogenic gel material with synergistic electro-bioactivity of the present invention comprises the following steps: S1. Add 0.01 to 0.15 parts of aqueous photoinitiator to 15 to 30 parts of PBS solution with a concentration of 0.01 mol / L, and stir at 40°C to 50°C for 5 to 20 minutes to obtain solution I; wherein the aqueous photoinitiator is Irgacure2959; S2. Add 0.5 to 10 parts of gelatin methyl propionate hydrogel to solution I, stir at 60℃ to 70℃ for 5 min to 20 min to obtain solution II; and sterilize solution II using a 0.22 μm sterile needle filter. S3. Add 0.2 to 5 parts of piezoelectric nanoparticles to solution II, and then sonicate for 10 to 30 minutes to obtain solution III, with an ultrasonic intensity of 0.3 W / cm. 2 ~0.6W / cm 2 ; S4. Add 0.6 to 15 parts of nanoporous bioglass to solution III, then sonicate for 10 to 30 minutes to obtain solution IV, with an ultrasonic intensity of 0.3 W / cm. 2 ~0.6W / cm2 ; S5. Place solution IV into a mold and irradiate it with an ultraviolet light source for 100s to 300s to obtain a highly efficient angiogenic gel material with synergistic electro-biological activity.

[0040] The nanoporous bioglass of the present invention is prepared by the following steps: A1. Dissolve hexadecyltrimethylammonium bromide in water to obtain solution A1; A2. Add ethyl acetate to solution A1 to obtain solution A2; A3. Add the ammonia solution to solution A2 to obtain solution A3; A4. Add tetraethyl orthosilicate to solution A3 to obtain solution A4; A5. Add calcium nitrate tetrahydrate to solution A4 to react, and then a precipitate is obtained; A6. The precipitate was calcined to obtain nanoporous bioglass.

[0041] The nanoporous bioglass, by weight, is prepared by the following steps: A1. Dissolve 1 to 4 parts of hexadecyltrimethylammonium bromide in 50 to 200 parts of water to obtain solution A1; A2. Add 20 to 60 parts of ethyl acetate to solution A1 to obtain solution A2; A3. Add 10 to 40 parts of 1 mol / L ammonia solution to solution A2 to obtain solution A3; A4. Add 5 to 25 parts of tetraethyl orthosilicate to solution A3 to obtain solution A4; A5. Add 2 to 20 parts of calcium nitrate tetrahydrate to solution A4 to react, and then obtain the precipitate. Use 200 to 300 parts of deionized water for each wash; use 200 to 300 parts of ethanol for each wash. A6. The precipitate was calcined to obtain nanoporous bioglass.

[0042] Furthermore, the nanoporous bioglass is prepared by the following steps: A1. Add hexadecyltrimethylammonium bromide to water and control the temperature to 30℃~40℃, stir for 10min~15min to obtain solution A1; A2. Add ethyl acetate to solution A1 and stir for 30 min to 40 min to obtain solution A2; A3. Add the ammonia solution to solution A2 and stir for 15 min to 20 min to obtain solution A3; A4. Add tetraethyl orthosilicate to solution A3 and stir for 30 min to 40 min to obtain solution A4; A5. Add calcium nitrate tetrahydrate to solution A4 and react for 4 to 6 hours. Then collect the solid and wash it several times with deionized water and ethanol alternately. Then dry it at 50 to 60 degrees Celsius for 12 to 18 hours to obtain the precipitate. A6. The precipitate was calcined at 600℃~900℃ and the sintering holding time was 2h~4h to obtain nanoporous bioglass.

[0043] This method for preparing a highly efficient angiogenic gel material with synergistic electro-bioactivity involves cross-linking gelatin methylpropionate hydrogel, piezoelectric nanoparticles, and mesoporous bioglass under the action of an aqueous photoinitiator. The mesoporous bioglass is prepared by emulsifying hexadecyltrimethylammonium bromide, tetraethyl orthosilicate, and a calcium source using a template method followed by high-temperature calcination. This invention offers the following advantages: 1. The preparation process of the highly efficient angiogenic gel material with synergistic electro-bioactivity is simple, stable, highly reproducible, and biocompatible. 2. This highly efficient angiogenic gel material can generate an electric field when subjected to external mechanical force, eliminating the need for an external power source and enhancing the mesoporous bioglass through a wireless field to achieve a highly efficient angiogenic effect. 3. The piezoelectric nanoparticles in this highly efficient angiogenic gel material not only provide the necessary electrical stimulation but also enhance the mechanical properties of the bulk hydrogel.

[0044] Example 2

[0045] A highly efficient angiogenic gel material with synergistic electro-bioactivity, having the same characteristics as in Example 1, except that: the nanoporous bioglass, by weight, is prepared by the following steps: A1. Add 1 part hexadecyltrimethylammonium bromide to 50 parts water, control the temperature to 30°C, and stir for 10 minutes to obtain solution A1; A2. Add 20 parts of ethyl acetate to solution A1 and stir for 30 minutes to obtain solution A2; A3. Add 40 parts of 1mol / L ammonia solution to solution A2 and stir for 20 min to obtain solution A3; A4. Add 5 parts of tetraethyl orthosilicate to solution A3 and stir for 40 minutes to obtain solution A4. A5. Add 20 parts of calcium nitrate tetrahydrate to solution A4 and react for 6 hours. Then collect the solid and wash it several times with deionized water and ethanol alternately. Then dry it at 50°C for 12 hours to obtain the precipitate. A6. The precipitate was calcined at 900℃ and the sintering holding time was 4h to obtain nanoporous bioglass.

[0046] The preparation method of the highly efficient angiogenic gel material with synergistic electro-bioactivity is carried out by the following steps, based on parts by weight: S1. Add 0.01 parts of aqueous photoinitiator to 30 parts of PBS solution and stir at 50°C for 5 min to obtain solution I; S2. Add 0.5 parts of gelatin methyl propionate hydrogel to solution I and stir at 70°C for 5 min to obtain solution II; S3. Add 0.2 parts of piezoelectric nanoparticles to solution II, then sonicate for 10 min to obtain solution III, with an ultrasonic intensity of 0.6 W / cm. 2 Solution II was sterilized using a 0.22 μm sterile needle filter; the piezoelectric nanoparticles were a mixture of bismuth tungstate and sodium bismuth titanate, with a weight ratio of bismuth tungstate to sodium bismuth titanate of 1:1, and were prepared by hydrothermal reaction with tetrabutyl titanate and barium hydroxide octahydrate. S4. Add 0.6 parts of nanoporous bioglass to solution III, then sonicate for 10 min to obtain solution IV, with an ultrasonic intensity of 0.3 W / cm. 2 ; S5. Place solution IV into a mold and irradiate it with an ultraviolet light source for 100s to obtain a highly efficient angiogenic gel material with synergistic electro-biological activity.

[0047] Example 3

[0048] A highly efficient angiogenic gel material with synergistic electro-bioactivity, having the same characteristics as in Example 1, except that: the nanoporous bioglass, by weight, is prepared by the following steps: A1. Add 4 parts of hexadecyltrimethylammonium bromide to 200 parts of water, and control the temperature to 40°C. Stir for 15 minutes to obtain solution A1. A2. Add 60 parts of ethyl acetate to solution A1 and stir for 40 minutes to obtain solution A2; A3. Add 10 parts of 1mol / L ammonia solution to solution A2 and stir for 15 min to obtain solution A3; A4. Add 25 parts of tetraethyl orthosilicate to solution A3 and stir for 30 minutes to obtain solution A4. A5. Add 2 parts of calcium nitrate tetrahydrate to solution A4 and react for 4 hours. Then collect the solid and wash it several times with deionized water and ethanol alternately. Then dry it at 60°C for 18 hours to obtain the precipitate. A6. The precipitate was calcined at 600℃ and the sintering holding time was 2h to obtain nanoporous bioglass.

[0049] The preparation method of the highly efficient angiogenic gel material with synergistic electro-bioactivity is carried out by the following steps, based on parts by weight: S1. Add 0.15 parts of aqueous photoinitiator to 15 parts of PBS solution and stir at 40°C for 20 min to obtain solution I; S2. Add 10 parts of gelatin methyl propionate hydrogel to solution I, stir at 70℃ for 20 min to obtain solution II; and sterilize solution II using a 0.22μm sterile needle filter; S3. Add 5 portions of piezoelectric nanoparticles to solution II, then sonicate for 30 min to obtain solution III, with an ultrasonic intensity of 0.3 W / cm. 2 The piezoelectric nanoparticles are potassium sodium niobate, which are prepared by hydrothermal reaction with tetrabutyl titanate and barium hydroxide octahydrate. S4. Add 15 portions of nanoporous bioglass to solution III, then sonicate for 30 min to obtain solution IV, with an ultrasonic intensity of 0.6 W / cm. 2 ; S5. Place solution IV into a mold and irradiate it with an ultraviolet light source for 300s to obtain a highly efficient angiogenic gel material with synergistic electro-biological activity.

[0050] Example 4

[0051] A highly efficient angiogenic gel material with synergistic electro-bioactivity, having the same characteristics as in Example 1, except that: the nanoporous bioglass, by weight, is prepared by the following steps: A1. Add 2 parts of hexadecyltrimethylammonium bromide to 150 parts of water, and control the temperature to 35°C. Stir for 13 minutes to obtain solution A1. A2. Add 25 parts of ethyl acetate to solution A1 and stir for 35 minutes to obtain solution A2; A3. Add 15 parts of 1 mol / L ammonia solution to solution A2 and stir for 17 min to obtain solution A3; A4. Add 10 parts of tetraethyl orthosilicate to solution A3 and stir for 35 minutes to obtain solution A4. A5. Add 10 parts of calcium nitrate tetrahydrate to solution A4 and react for 5 hours. Then collect the solid and wash it several times with deionized water and ethanol alternately. Then dry it at 55°C for 16 hours to obtain the precipitate. A6. The precipitate was calcined at 720℃ and the sintering holding time was 3h to obtain nanoporous bioglass.

[0052] The preparation method of the highly efficient angiogenic gel material with synergistic electro-bioactivity is carried out by the following steps, based on parts by weight: S1. Add 0.08 parts of aqueous photoinitiator to 20 parts of PBS solution and stir at 40°C for 10 min to obtain solution I; S2. Add 2 portions of gelatin methyl propionate hydrogel to solution I and stir at 60°C for 15 min to obtain solution II; sterilize solution II using a 0.22 μm sterile needle filter; S3. Add two portions of piezoelectric nanoparticles to solution II, then sonicate for 10 min to obtain solution III, with an ultrasonic intensity of 0.5 W / cm. 2 ; S4. Add two portions of nanoporous bioglass to solution III, then sonicate for 20 min to obtain solution IV, with an ultrasonic intensity of 0.4 W / cm. 2 The piezoelectric nanoparticles are barium titanate, which are prepared by hydrothermal reaction with tetrabutyl titanate and barium hydroxide octahydrate. S5. Place solution IV into a mold and irradiate it with an ultraviolet light source for 200s to obtain a highly efficient angiogenic gel material with synergistic electro-biological activity.

[0053] Example 5

[0054] A highly efficient angiogenic gel material with synergistic electro-bioactivity, having the same characteristics as in Example 1, except that: the nanoporous bioglass, by weight, is prepared by the following steps: A1. Add 2 parts of hexadecyltrimethylammonium bromide to 150 parts of water, and control the temperature to 35°C. Stir for 13 minutes to obtain solution A1. A2. Add 20 parts of ethyl acetate to solution A1 and stir for 35 minutes to obtain solution A2; A3. Add 15 parts of 1mol / L ammonia solution to solution A2 and stir for 18 min to obtain solution A3; A4. Add 15 parts of tetraethyl orthosilicate to solution A3 and stir for 34 min to obtain solution A4; A5. Add 15 parts of calcium nitrate tetrahydrate to solution A4 and react for 5 hours. Then collect the solid and wash it several times with deionized water and ethanol alternately. Then dry it at 55°C for 16 hours to obtain the precipitate. A6. The precipitate was calcined at 800℃ and the sintering holding time was 3h to obtain nanoporous bioglass.

[0055] The preparation method of the highly efficient angiogenic gel material with synergistic electro-bioactivity is carried out by the following steps, based on parts by weight: S1. Add 0.01 parts of aqueous photoinitiator to 20 parts of PBS solution and stir at 45°C for 10 min to obtain solution I; S2. Add 5 parts of gelatin methyl propionate hydrogel to solution I, stir at 63℃ for 10 min to obtain solution II; and sterilize solution II using a 0.22μm sterile needle filter; S3. Add one part of piezoelectric nanoparticles to solution II, then sonicate for 20 min to obtain solution III, with an ultrasonic intensity of 0.5 W / cm. 2 The piezoelectric nanoparticles are barium titanate, which are prepared by hydrothermal reaction with tetrabutyl titanate and barium hydroxide octahydrate. S4. Add 3 portions of nanoporous bioglass to solution III, then sonicate for 20 min to obtain solution IV, with an ultrasonic intensity of 0.4 W / cm. 2 ; S5. Place solution IV into a mold and irradiate it with an ultraviolet light source for 150s to obtain a highly efficient angiogenic gel material with synergistic electro-biological activity.

[0056] Comparative Example 1

[0057] A highly efficient angiogenic gel material with synergistic electro-bioactivity, having the same characteristics as in Example 5, except that, by weight, the preparation method of the highly efficient angiogenic gel material with synergistic electro-bioactivity comprises the following steps: S1. Add 0.01 parts of aqueous photoinitiator to 20 parts of PBS solution and stir at 40℃~50℃ for 5min~20min to obtain solution I; S2. Add 5 parts of gelatin methyl propionate hydrogel to solution I, stir at 63℃ for 10 min to obtain solution II; and sterilize solution II using a 0.22μm sterile needle filter; S3. Add one part of piezoelectric nanoparticles to solution II, then sonicate for 20 min to obtain solution III, with an ultrasonic intensity of 0.5 W / cm. 2 The piezoelectric nanoparticles are barium titanate, which are prepared by hydrothermal reaction with tetrabutyl titanate and barium hydroxide octahydrate. S4. Place solution III into a mold and irradiate it with an ultraviolet light source for 150s to obtain a highly efficient angiogenic gel material with synergistic electro-biological activity.

[0058] Comparative Example 2

[0059] A highly efficient angiogenic gel material with synergistic electro-bioactivity, which shares the same characteristics as in Example 5, except that, by weight, the nanoporous bioglass is prepared by the following steps: A1. Add 6 parts of hexadecyltrimethylammonium bromide to 200 parts of water, control the temperature to 30°C, and stir for 10 minutes to obtain solution A1; A2. Add 5 parts of ethyl acetate to solution A1 and stir for 40 min to obtain solution A2; A3. Add 30 parts of 1mol / L ammonia solution to solution A2 and stir for 15min to 20min to obtain solution A3; A4. Add 2 parts of tetraethyl orthosilicate to solution A3 and stir for 40 minutes to obtain solution A4; A5. Add 40 parts of calcium nitrate tetrahydrate to solution A4 and react for 6 hours. Then collect the solid and wash it several times with deionized water and ethanol alternately. Then dry it at 50°C for 18 hours to obtain the precipitate. A6. The precipitate was calcined at 800℃ and the sintering holding time was 5h to obtain nanoporous bioglass.

[0060] Comparative Example 3

[0061] A highly efficient angiogenic gel material with synergistic electro-bioactivity, having the same characteristics as in Example 5, except that, by weight, the preparation method of the highly efficient angiogenic gel material with synergistic electro-bioactivity comprises the following steps: S1. Add 0.03 parts of aqueous photoinitiator to 20 parts of PBS solution and stir at 44℃ for 10 min to obtain solution I; S2. Add 15 parts of gelatin methyl propionate hydrogel to solution I, stir at 60℃ for 10 min to obtain solution II; and sterilize solution II using a 0.22μm sterile needle filter; S3. Add 6 portions of piezoelectric nanoparticles to solution II, then sonicate for 10-30 minutes to obtain solution III, with an ultrasonic intensity of 0.4 W / cm. 2 ; S4. Add 0.1 part of nanoporous bioglass to solution III, then sonicate for 20 min to obtain solution IV, with an ultrasonic intensity of 0.5 W / cm. 2 ; S5. Place solution IV into a mold and irradiate it with an ultraviolet light source for 150s to obtain a highly efficient angiogenic gel material with synergistic electro-biological activity.

[0062] Test case

[0063] 1. Morphological and elemental analysis of nanoporous bioglass

[0064] The nanoporous bioglasses prepared in Examples 4 and 5 were subjected to microscopic and elemental analysis to obtain the corresponding results. Figure 1 and Figure 2 .

[0065] in Figure 2 Changing the amount of ethyl acetate added, as well as altering the silicon source and the proportions of other components, significantly affects the morphology and silicon / calcium content of the nanoporous bioglass. Figure 1 and Figure 2Observed under a field emission scanning electron microscope, the nanoporous bioglass exhibits a clear mesoporous structure and uniform particle distribution, with no obvious agglomeration. It can be seen that the nanoporous bioglass obtained by this invention can be used to prepare mesoporous materials with regular structures.

[0066] 2. Microstructure analysis of piezoelectric materials

[0067] The piezoelectric material prepared in Example 5 was analyzed under a microscope, and the corresponding results were obtained. Figure 3 .

[0068] pass Figure 3 As can be seen from the field emission scanning electron microscope, the piezoelectric material of this invention consists of nanoscale particles with good dispersion and no obvious agglomeration. The nanoscale size allows the piezoelectric particles to be uniformly dispersed in the gel matrix, ensuring the uniform generation of the electric field when the gel material is subjected to mechanical force, while also improving the binding with the gel matrix and avoiding the decline in mechanical properties caused by agglomeration. The uniform size of the piezoelectric nanoparticles can be observed from the morphology.

[0069] 3. Macroscopic analysis of highly efficient angiogenic gel materials with synergistic electro-bioactivity.

[0070] The electro-bioactive synergistic high-efficiency angiogenesis-promoting gel materials prepared in Examples 4 and 5 were subjected to appearance analysis, and the corresponding results were obtained. Figure 4 ,in Figure 4 A represents the highly efficient angiogenesis-promoting gel material obtained in Example 4, characterized by synergistic electro-bioactivity. Figure 4 B is the highly efficient angiogenic gel material with synergistic electro-bioactivity obtained in Example 5.

[0071] from Figure 4 As can be seen, the highly efficient vascular gel materials of the present invention all exhibit a uniform gel morphology, without layering or particle precipitation, proving that gelatin methyl propionate hydrogel, piezoelectric nanoparticles, and nanoporous bioglass have achieved effective composite cross-linking under the action of aqueous photoinitiator, with each component uniformly dispersed in the gel matrix and good cross-linking molding effect.

[0072] 4. Micromorphology analysis of highly efficient angiogenic gel materials with synergistic electro-bioactivity.

[0073] Microscopic analysis of the highly efficient angiogenic gel materials with synergistic electro-bioactivity prepared in Examples 4 and 5 yielded the corresponding results. Figure 5 and Figure 6 .

[0074] pass Figure 5 and Figure 6As can be seen, the highly efficient vascular-promoting gel material obtained in this invention exhibits a three-dimensional porous cross-linked network structure under field emission scanning electron microscopy, with uniform pore distribution. This structure is key to the hydrogel's water retention and air permeability, providing space for the adhesion and proliferation of vascular endothelial cells, while facilitating the exchange of nutrients and metabolites, thus meeting the microenvironmental requirements for skin wound repair. Moreover, there are no obvious particle aggregates in the microstructure, proving that the piezoelectric nanoparticles and nanoporous bioglass are uniformly dispersed in the gel network, without disrupting the three-dimensional network structure of the gel due to particle addition. Furthermore, the uniform particle dispersion ensures consistent electroactivity and biological activity across all parts of the gel material, avoiding local performance deficiencies.

[0075] 5. Highly efficient angiogenic gel materials with synergistic electro-bioactivity promote angiogenesis: fluorescence analysis of angiogenesis.

[0076] This experimental design uses human umbilical vein endothelial cells (HUVECs), P5 generation, as the research subject. Angiogenic fluorescence images were obtained using a matrix tubing formation assay. Figure 7 .

[0077] Before the experiment, complete endothelial cell culture medium, Matrigel, and Calcein-AM reagent (a live cell dye) were prepared. HUVECs were cultured to a suitable concentration and a cell suspension with a cell density of 10⁵ cells / mL was prepared. Matrigel and experimental materials were pre-cooled to 4°C, and the samples were then incubated at 37°C for 1 hour to promote solidification. The prepared cell suspension was seeded onto the surface of the electro-bioactive synergistic high-efficiency angiogenesis-promoting gel material prepared in the blank group, Examples 2 to 5, and Comparative Examples 1 to 3, with a cell seeding density of 5 × 10⁶ cells / mL. 4 Each well contains 48 cells, which are cultured for 12 hours. Finally, the cells are stained with 1 mg / mL calcein AM for 15 minutes, and then the number of vascular branches is analyzed using ImageJ software, as shown in Table 1.

[0078] Table 1. Data on the angiogenesis-promoting effects of highly efficient angiogenic gel materials

[0079] Table 1 shows that the blank group had 10 vascular branches. The electro-bioactive synergistic gel systems of Examples 2-5 had 40, 42, 44, and 45 vascular branches respectively, showing a gradually increasing gradient trend. This demonstrates the significant synergistic effect of the nanoporous bioglass and piezoelectric nanoparticles, resulting in pro-angiogenic performance far superior to a single electroactive system. Example 5 represents the optimal overall formulation, achieving peak pro-angiogenic performance, with a value 350% higher than the blank group, fully demonstrating the highly efficient pro-angiogenic properties of the gel material of this invention. The number of vascular branches in Comparative Examples 1-3 were 36, 36, and 38 respectively, all significantly lower than in Examples 2-5. Comparative Example 1 lacked nanoporous bioglass, verifying that nanoporous bioglass is the core for achieving electro-bioactive synergy. The nanoporous bioglass formulation of Comparative Example 2 and the overall gel material formulation of Comparative Example 3 both exceeded the formulation limits of this invention, resulting in extremely poor pro-angiogenic performance.

[0080] 6. Hemolysis analysis of highly efficient angiogenic gel materials with synergistic electro-bioactivity.

[0081] The positive control Triton and the electro-bioactive synergistic highly efficient angiogenic gel materials prepared in Examples 2-5 were co-incubated with erythrocyte suspension (37°C, 1 h). After centrifugation, the color of the supernatant and the erythrocyte sedimentation were observed. Figure 8 ,exist Figure 8 From left to right: positive control, Example 2, Example 3, Example 4, and Example 5.

[0082] from Figure 8 As can be seen, the supernatant of the positive control was bright red and showed no red blood cell precipitation, indicating complete hemolysis and verifying the reliability of the experimental system. The supernatant of the highly efficient angiogenic gel material with synergistic electro-biological activity in Examples 2-5 was colorless and transparent, with intact red blood cell precipitation at the bottom of the tube, indicating that these four samples had no significant hemolytic effect under the experimental conditions, and the degree of hemolysis was extremely low, far lower than that of the positive control group, demonstrating good blood compatibility from the perspective of hemolysis.

[0083] Example 6

[0084] The use of a highly efficient angiogenic gel material with synergistic electro-bioactivity in the preparation of deep subcutaneous tissue skin wound repair products. The highly efficient angiogenic gel material with synergistic electro-bioactivity is prepared by the preparation methods of the highly efficient angiogenic gel material with synergistic electro-bioactivity in Examples 1 to 5.

[0085] The test data show that the highly efficient angiogenesis-promoting gel material of the present invention can effectively promote the generation of blood vessels compared with the comparative example. Therefore, it can be used in biomedical materials for skin wound repair, especially suitable for the repair of deep subcutaneous tissue skin wounds with damaged vascular networks, filling the gap in the prior art.

[0086] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A method for preparing a highly efficient angiogenic gel material with synergistic electro-bioactivity, characterized in that: The gel was prepared by cross-linking gelatin methyl propionate hydrogel, piezoelectric nanoparticles and nanoporous bioglass under the action of an aqueous photoinitiator. The nanoporous bioglass is prepared by emulsification of hexadecyltrimethylammonium bromide, tetraethyl orthosilicate and calcium source using a template method followed by high-temperature calcination.

2. The method for preparing the highly efficient angiogenic gel material with synergistic electro-bioactivity according to claim 1, characterized in that, The nanoporous bioglass is prepared by the following steps: A1. Dissolve hexadecyltrimethylammonium bromide in water to obtain solution A1; A2. Add ethyl acetate to solution A1 to obtain solution A2; A3. Add the ammonia solution to solution A2 to obtain solution A3; A4. Add tetraethyl orthosilicate to solution A3 to obtain solution A4; A5. Add calcium nitrate tetrahydrate to solution A4 to react, and then a precipitate is obtained; A6. The precipitate is calcined to obtain the nanoporous bioglass.

3. The method for preparing the highly efficient angiogenic gel material with synergistic electro-bioactivity according to claim 2, characterized in that, The nanoporous bioglass, by weight, is prepared by the following steps: A1. Dissolve 1 to 4 parts of hexadecyltrimethylammonium bromide in 50 to 200 parts of water to obtain solution A1; A2. Add 20 to 60 parts of ethyl acetate to solution A1 to obtain solution A2; A3. Add 10 to 40 parts of 1 mol / L ammonia solution to solution A2 to obtain solution A3; A4. Add 5 to 25 parts of tetraethyl orthosilicate to solution A3 to obtain solution A4; A5. Add 2 to 20 parts of calcium nitrate tetrahydrate to solution A4 and react to obtain a precipitate; A6. The precipitate is calcined to obtain the nanoporous bioglass.

4. The method for preparing the highly efficient angiogenic gel material with synergistic electro-bioactivity according to claim 3, characterized in that, The nanoporous bioglass is prepared by the following steps: A1. Add hexadecyltrimethylammonium bromide to water and control the temperature to 30℃~40℃, stir for 10min~15min to obtain solution A1; A2. Add ethyl acetate to solution A1 and stir for 30 min to 40 min to obtain solution A2; A3. Add the ammonia solution to solution A2 and stir for 15 min to 20 min to obtain solution A3; A4. Add tetraethyl orthosilicate to solution A3 and stir for 30 min to 40 min to obtain solution A4; A5. Add calcium nitrate tetrahydrate to solution A4 and react for 4 to 6 hours. Then collect the solid and wash it several times with deionized water and ethanol alternately. Then dry it at 50°C to 60°C for 12 to 18 hours to obtain the precipitate. A6. The precipitate is calcined at 600℃~900℃ and the sintering holding time is 2h~4h to obtain the nanoporous bioglass.

5. The method for preparing the highly efficient angiogenic gel material with synergistic electro-bioactivity according to claim 4, characterized in that: In the A5, the amount of deionized water used for each cleaning is 200 to 300 parts; the amount of ethanol used for each cleaning is 200 to 300 parts.

6. The method for preparing the highly efficient angiogenic gel material with synergistic electro-bioactivity according to any one of claims 1 to 5, characterized in that, The procedure, by weight, is as follows: S1. Add 0.01 to 0.15 parts of aqueous photoinitiator to 15 to 30 parts of PBS solution to obtain solution I; S2. Add 0.5 to 10 parts of gelatin methyl propionate hydrogel to solution I to obtain solution II; S3. Add 0.2 to 5 parts of piezoelectric nanoparticles to solution II, and then treat with ultrasound to obtain solution III; S4. Add 0.6 to 15 parts of nanoporous bioglass to solution III, and then treat with ultrasound to obtain solution IV; S5. Solution IV is placed in a mold and irradiated with an ultraviolet light source to obtain a highly efficient angiogenic gel material with synergistic electro-biological activity.

7. The method for preparing the highly efficient angiogenic gel material with synergistic electro-bioactivity according to claim 6, characterized in that, The process is as follows: S1. Add 0.01 to 0.15 parts of aqueous photoinitiator to 15 to 30 parts of PBS solution and stir at 40°C to 50°C for 5 to 20 minutes to obtain solution I; S2. Add 0.5 to 10 parts of gelatin methyl propionate hydrogel to solution I and stir at 60℃ to 70℃ for 5 min to 20 min to obtain solution II; S3, 0.2-5 parts of piezoelectric nanoparticles are added to solution II, and then ultrasonic treatment is performed for 10-30 min to obtain solution III, and the ultrasonic intensity is 0.3-0.6 W / cm 2 . 2 ​ S4. Add 0.6 to 15 parts of nanoporous bioglass to solution III, then sonicate for 10 to 30 minutes to obtain solution IV, with an ultrasonic intensity of 0.3 W / cm. 2 ~0.6W / cm 2 ; S5. Place solution IV into a mold and irradiate it with an ultraviolet light source for 100s to 300s to obtain a highly efficient angiogenic gel material with synergistic electro-biological activity.

8. The method for preparing the highly efficient angiogenic gel material with synergistic electro-bioactivity according to any one of claims 1 to 5, characterized in that, The calcium source is calcium nitrate tetrahydrate; The gelatin methyl propionate hydrogel was prepared from methacrylic anhydride-modified gelatin. The piezoelectric nanoparticles are one or more of barium titanate, potassium sodium niobate, bismuth tungstate, and sodium bismuth titanate, and are prepared by hydrothermal reaction with tetrabutyl titanate and barium hydroxide octahydrate.

9. A highly efficient angiogenic gel material with synergistic electro-bioactivity, characterized in that: It is prepared by the method for preparing a highly efficient vascular-promoting gel material with synergistic electro-bioactivity as described in any one of claims 1 to 8.

10. The use of a highly efficient angiogenic gel material with synergistic electro-bioactivity in the preparation of deep subcutaneous tissue skin wound repair products, characterized in that: The electro-bioactive synergistic highly efficient angiogenic gel material is prepared by the preparation method of the electro-bioactive synergistic highly efficient angiogenic gel material according to any one of claims 1 to 8.

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