阅读说明：本技术 抗菌消炎复合水凝胶前驱液及其制备方法和应用 (Antibacterial and anti-inflammatory composite hydrogel precursor solution and preparation method and application thereof ) 是由 潘浩波 崔旭 庞力斌 田鹏飞 于 2021-09-13 设计创作，主要内容包括：本申请提供了一种抗菌消炎复合水凝胶前驱液,包括：甲基丙烯酰氧化丝素蛋白、生物活性玻璃、光引发剂和水,其中,甲基丙烯酰氧化丝素蛋白的分子量为10000Da-15000Da。该抗菌消炎复合水凝胶前驱液经光固化后能够形成抗菌消炎复合水凝胶,该复合水凝胶不仅具有良好的保湿能力,并且还具有抑菌、促进血管新生和组织生长的功能,从而促进患者创面的再生修复,缩短创面的愈合时间,达到良好的治疗效果。本申请还提供了一种抗菌消炎复合水凝胶前驱液的制备方法和应用。(The application provides an antibacterial and anti-inflammatory composite hydrogel precursor liquid, which comprises: the silk fibroin material comprises methacryloyl oxidation silk fibroin, bioactive glass, a photoinitiator and water, wherein the molecular weight of the methacryloyl oxidation silk fibroin is 10000Da-15000 Da. The antibacterial and anti-inflammatory composite hydrogel precursor liquid can form the antibacterial and anti-inflammatory composite hydrogel after photocuring, and the composite hydrogel not only has good moisturizing capability, but also has the functions of inhibiting bacteria and promoting angiogenesis and tissue growth, so that the regenerative repair of the wound surface of a patient is promoted, the healing time of the wound surface is shortened, and a good treatment effect is achieved. The application also provides a preparation method and application of the antibacterial and anti-inflammatory composite hydrogel precursor liquid.)

1. An antibacterial and anti-inflammatory composite hydrogel precursor solution, which is characterized by comprising: the biological glass comprises methacryl oxidized silk fibroin, biological active glass, a photoinitiator and water; the molecular weight of the methacryl oxidized silk fibroin is 10000Da-15000 Da.

2. The antibacterial and anti-inflammatory composite hydrogel precursor solution as claimed in claim 1, wherein the bioactive glass comprises the following components in mol percentage: SiO 2₂：20％-80％；B₂O₃: 5% -40%; XO: 15% -40%, wherein the X comprises one or more of Ca, Mg, Sr and Cu.

3. The antimicrobial and anti-inflammatory composite hydrogel precursor of claim 2, wherein the XO comprises CaO, MgO, SrO, and CuO; the mol percentages of the CaO, the MgO, the SrO and the CuO in the bioactive glass are respectively 15-25%, 5-10%, 3-8% and 0.1-2%.

4. The antimicrobial and anti-inflammatory composite hydrogel precursor solution of claim 2 or 3, wherein the bioactive glass further comprises P₂O₅Said P is₂O₅The molar percentage in the bioactive glass is less than or equal to 4%.

5. The antibacterial and anti-inflammatory composite hydrogel precursor solution according to any one of claims 1 to 4, wherein the bioactive glass is grafted with a methacryloxy group, and the methacryloxy group is connected with the bioactive glass through a siloxane bond; the particle size of the bioactive glass is 0.5-5 μm.

6. The antibacterial and anti-inflammatory composite hydrogel precursor solution as claimed in any one of claims 1 to 5, wherein the methacryloxy-modified silk fibroin is silk fibroin modified with methacryloxy groups, and the mass percentage of the methacryloxy groups in the methacryloxy-modified silk fibroin is 20% to 40%; the photoinitiator comprises lithium phenyl-2, 4, 6-trimethylbenzoyl phosphate.

7. The antibacterial and anti-inflammatory composite hydrogel precursor solution as claimed in any one of claims 1 to 6, wherein the antibacterial and anti-inflammatory composite hydrogel precursor solution comprises the following components in percentage by mass: methacryloxy oxidized silk fibroin: 10% -30%; bioactive glass: 1% -6%; photoinitiator (2): 0.1% -0.4%; water: 64 to 88 percent.

8. The antibacterial and anti-inflammatory composite hydrogel precursor solution according to any one of claims 1 to 7, wherein the antibacterial and anti-inflammatory composite hydrogel precursor solution is cured under ultraviolet irradiation to form an antibacterial and anti-inflammatory composite hydrogel; the tensile strength of the composite hydrogel is 20Kpa-40 Kpa.

9. The anti-microbial, anti-inflammatory composite hydrogel precursor liquid of any one of claims 1 to 8, further comprising one or more of an anti-inflammatory drug, an anti-microbial drug, and a growth factor drug.

10. A preparation method of an antibacterial and anti-inflammatory composite hydrogel precursor solution is characterized by comprising the following steps: mixing methacryloyl oxidized silk fibroin, bioactive glass, a photoinitiator and water to obtain an antibacterial and anti-inflammatory composite hydrogel precursor solution; the molecular weight of the methacryl oxidized silk fibroin is 10000Da-15000 Da.

11. A dressing prepared from the antibacterial anti-inflammatory composite hydrogel precursor solution of any one of claims 1 to 9.

Technical Field

The application relates to the technical field of biomedical materials, in particular to an antibacterial and anti-inflammatory composite hydrogel precursor liquid and a preparation method and application thereof.

Background

The chronic wound surface of diabetes is one of the common clinical diseases, because the organism metabolic disturbance, the immunologic function are damaged, the wound surface is difficult to regenerate and repair, and the high sugar environment in the body of the diabetes is easy to cause bacterial infection and aggravate inflammatory reaction, the difficulty of wound surface repair is further deepened.

Traditional dressings such as gauze and linen have a certain protective effect on wounds, however, because the traditional dressings have no moisture retention and need to be replaced for many times, the wounds are easy to adhere in the treatment process, the treatment period is long, the secondary damage of the wounds is easy to cause, and the repair of the wound surface is not facilitated. Therefore, there is a need to develop an external dressing capable of resisting bacteria, diminishing inflammation and inducing regeneration of human tissues to promote the regenerative repair of the wound surface of a patient.

Disclosure of Invention

In order to solve the problems, the application provides an antibacterial and anti-inflammatory composite hydrogel precursor solution, the antibacterial and anti-inflammatory composite hydrogel precursor solution can form an antibacterial and anti-inflammatory composite hydrogel after photocuring, and the composite hydrogel not only has good moisturizing capability, but also has the functions of inhibiting bacteria, promoting angiogenesis and tissue growth, thereby promoting the regenerative repair of the wound surface of a patient, shortening the healing time of the wound surface and achieving a good treatment effect.

Specifically, the application provides, in a first aspect, an antibacterial and anti-inflammatory composite hydrogel precursor solution, which comprises: the biological glass comprises methacryl oxidized silk fibroin, biological active glass, a photoinitiator and water; the molecular weight of the methacryl oxidized silk fibroin is 10000Da-15000 Da.

In the antibacterial and anti-inflammatory composite hydrogel precursor solution, the methacryl oxidized silk fibroin can be quickly crosslinked under the action of a photoinitiator and ultraviolet light to form the composite hydrogel, the obtained composite hydrogel has good mechanical strength so as to effectively protect a wound surface, and the molecular weight of the methacryl oxidized silk fibroin is controlled so as to ensure that the composite hydrogel has good moisture retention performance and reduce the stimulation to the wound in the composite hydrogel replacement process; the bioactive glass has the functions of resisting bacteria and diminishing inflammation, and the methacryl oxidized silk fibroin and the bioactive glass have good compatibility, so the bioactive glass can be uniformly dispersed in the composite hydrogel, the composite hydrogel can reduce the inflammation of a wound surface, and the wound surface can be effectively repaired.

Optionally, the bioactive glass comprises the following components in percentage by mole: SiO 2₂：20％-80％；B₂O₃: 5% -40%; XO: 15% -40%, wherein the X comprises one or more of Ca, Mg, Sr and Cu.

Optionally, the XO includes CaO, MgO, SrO, and CuO; the mol percentages of the CaO, the MgO, the SrO and the CuO in the bioactive glass are respectively 15-25%, 5-10%, 3-8% and 0.1-2%.

Optionally, the bioactive glass further comprises P₂O₅Said P is₂O₅The molar percentage in the bioactive glass is less than or equal to 4%.

Optionally, the bioactive glass is grafted with a methacryloxy group; the methacryloxy group is attached to the bioactive glass via a siloxane bond.

Optionally, the bioactive glass has a particle size of 0.5 μm to 5 μm.

Optionally, the methacryloxy-modified silk fibroin is modified with methacryloxy groups, and the mass percentage of the methacryloxy groups in the methacryloxy-modified silk fibroin is 20% -40%.

Optionally, the photoinitiator comprises lithium phenyl-2, 4, 6-trimethylbenzoyl phosphate.

Optionally, the antibacterial and anti-inflammatory composite hydrogel precursor solution comprises the following components in percentage by mass: methacryloxy oxidized silk fibroin: 10% -30%; bioactive glass: 1% -6%; photoinitiator (2): 0.1% -0.4%; water: 64 to 88 percent.

Optionally, the antibacterial and anti-inflammatory composite hydrogel precursor solution is cured under ultraviolet irradiation to form an antibacterial and anti-inflammatory composite hydrogel; the tensile strength of the composite hydrogel is 20Kpa-40 Kpa.

Optionally, the antibacterial and anti-inflammatory composite hydrogel precursor solution further comprises one or more of an anti-inflammatory drug, an antibacterial drug and a growth factor drug.

Optionally, mixing the methacryl oxidized silk fibroin, the bioactive glass, the photoinitiator and water to obtain an antibacterial and anti-inflammatory composite hydrogel precursor solution; the molecular weight of the methacryl oxidized silk fibroin is 10000Da-15000 Da.

The second aspect of the application provides a preparation method of an antibacterial and anti-inflammatory composite hydrogel precursor solution, which comprises the following steps:

mixing methacryloyl oxidized silk fibroin, bioactive glass, a photoinitiator and water to obtain an antibacterial and anti-inflammatory composite hydrogel precursor solution; the molecular weight of the methacryl oxidized silk fibroin is 10000Da-15000 Da.

In a third aspect, the present application provides a dressing prepared from the antibacterial and anti-inflammatory composite hydrogel precursor solution according to the first aspect of the present application.

Optionally, the preparation method of the dressing comprises: and (3) carrying out ultraviolet irradiation on the antibacterial and anti-inflammatory composite hydrogel precursor liquid to obtain the dressing.

Optionally, the wavelength of the ultraviolet light is 315nm to 400 nm.

Optionally, the irradiation time of the ultraviolet light is 10s-60 s.

Drawings

FIG. 1 is a nuclear magnetic resonance spectrum of methacryloylated oxidized silk fibroin (SF-MA) prepared in example 1;

FIG. 2 is a graph showing the particle size distribution of the bioactive glass obtained in example 2;

FIG. 3 is an X-ray diffraction pattern of the bioactive glass prepared in example 3;

FIG. 4 is an infrared characterization chart of Silk Fibroin (SF), methacryloyloxyl silk fibroin (SF-MA), Bioactive Glass (BG), methacryloyloxyl bioactive glass (BG-MA), and antibacterial and anti-inflammatory composite hydrogel (SF-MA-BG) prepared in example 3;

FIG. 5 is an electron microscope characterization chart of the dried antibacterial and anti-inflammatory composite hydrogel provided in example 3;

FIG. 6 is a graph showing swelling property test of the antibacterial and anti-inflammatory composite hydrogels of examples 3 to 5 and the hydrogel (SF-MA) of comparative example 1;

FIG. 7 is a graph showing mechanical properties of the antibacterial and anti-inflammatory composite hydrogels of examples 3 to 5 and the hydrogel (SF-MA) of comparative example 1;

FIG. 8 is a graph showing the wound healing effects of control group 1, control group 2 and example 4;

fig. 9 is a graph showing the results of H & E staining of the wound surfaces of control group 1, control group 2 and example 4.

Detailed Description

The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is obvious that the described embodiments are only a part of the embodiments of the present application, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.

Clinical treatment of chronic wounds of diabetes still faces a huge challenge, firstly, the chronic damage and dysfunction of many tissues, especially blood vessels, can be caused by the long-standing hyperglycemic environment in the body of a diabetic patient; secondly, under the state of diabetes, macrophage function and phenotype transformation are disordered to form continuously expanded inflammatory reaction so as to hinder the regeneration and repair of the wound surface; finally, the high sugar environment easily causes bacterial infection, aggravates inflammatory reaction and causes the wound surface of the diabetes to be difficult to heal. In order to obtain the dressing with good antibacterial and anti-inflammatory properties, the application provides the antibacterial and anti-inflammatory composite hydrogel precursor solution, after the precursor solution is covered on a wound surface, the precursor solution can be rapidly crosslinked and cured under the irradiation of ultraviolet light to form the composite hydrogel, the composite hydrogel can be in close contact with the wound surface, the wound surface is effectively isolated from the external environment, the invasion of bacteria is prevented, the composite hydrogel has a certain antibacterial and anti-inflammatory function, the immune environment can be regulated and controlled, the regeneration of local blood vessels and tissues is promoted, and therefore the healing of the diabetic wound surface is accelerated finally.

In the embodiment of the application, the antibacterial and anti-inflammatory composite hydrogel precursor solution comprises: the silk fibroin material comprises methacryl oxidized silk fibroin, bioactive glass, a photoinitiator and water. Wherein, methacryloxy oxidized silk fibroin (SF-MA) refers to silk fibroin modified with methacryloxy groups. Silk fibroin has good biocompatibility and skin tissue structure bionic property, and is beneficial to application of the silk fibroin in wound dressing, however, the gelling property of the silk fibroin is poor, and the formed hydrogel has low mechanical strength and cannot effectively protect the wound, so that the silk fibroin is modified by the application, the curing speed of the hydrogel can be accelerated by grafting the silk fibroin with the methacryloxy group, and the mechanical strength of the hydrogel can be effectively improved.

In the embodiment of the application, the methacryl oxidized silk fibroin is obtained by reacting Glycidyl Methacrylate (GMA) with an amino group on Silk Fibroin (SF) through an epoxy group, and the structural formula of the methacryl oxidized silk fibroin is as follows:

wherein the polymerization degree n is 15-25.

In some embodiments of the present disclosure, the methacryloxy group accounts for 20% to 40% by mass of the methacryl oxidized silk fibroin. Too high content of methacryloxy in the methacryloxy silk fibroin can cause less active amino in the silk fibroin and reduce the biocompatibility of the composite hydrogel; too low content of methacryloxy groups can result in long time for crosslinking and curing the precursor solution to form the composite hydrogel and poor mechanical properties of the composite hydrogel.

In the embodiment of the application, the molecular weight of the methacryl oxidized silk fibroin (SF-MA) is 10000Da-15000Da, and the molecular weight of the methacryl oxidized silk fibroin (SF-MA) can be specifically but not limited to 10000Da, 12000Da, 13000Da, 14000Da or 15000 Da. The molecular weight of SF-MA is controlled to ensure that the composite hydrogel formed by the precursor liquid has good lubricity and moist feeling, so that the friction force between the composite hydrogel and a wound surface can be reduced, the pain of a patient is relieved, and the composite hydrogel can effectively block the water loss of epidermal cells to play a role in protecting the skin. In the embodiment of the application, the mass percentage of the methacryl oxidized silk fibroin in the antibacterial and anti-inflammatory composite hydrogel precursor solution is 10% -30%. The mass percentage of the methacryl oxidized silk fibroin in the antibacterial and anti-inflammatory composite hydrogel precursor solution can be, but is not limited to, 10%, 15%, 20% or 30%.

In the embodiment of the application, the bioactive glass comprises the following components in percentage by mole: SiO 2₂：20％-80％；B₂O₃: 5% -40%; XO: 15% -40%, wherein X comprises one or more of Ca, Mg, Sr and Cu. The bioactive glass has the functions of resisting bacteria and diminishing inflammation, and can improve the structural strength of the composite hydrogel and avoid the damage of the dressing, so that the wound surface is effectively protected.

In the embodiments of the present application, SiO₂The mol percentage of the bioactive glass is 20-80%. SiO 2₂The molar percentage in the bioactive glass may in particular be, but not limited to, 20%, 30%, 40%, 50%, 60% or 80%. SiO in bioactive glass₂Can promote angiogenesis, and is made of SiO₂Too high a content of (b) can reduce the bioactivity of the composite hydrogel, and is not beneficial to wound repair. In the embodiments of the present application, B₂O₃The mol percentage of the bioactive glass is 5 to 40 percent. B is₂O₃The molar percentage in the bioactive glass may in particular be, but not limited to, 5%, 10%, 20%, 30% or 40%. B in bioactive glass₂O₃Can effectively regulate and control the growth ofDegradation Properties of bioactive glass, B₂O₃Too high content of (A) can result in too high activity of the bioglass, and the B element is precipitated too fast to cause certain toxicity.

In an embodiment of the present application, XO includes one or more of CaO, MgO, SrO, and CuO. XO can decompose X in the composite hydrogel²⁺For example, when XO is CaO, X²⁺Is Ca²⁺(ii) a When XO is SrO, X²⁺Is Sr²⁺。X²⁺The functional ion can regulate and control inflammation, inhibit bacterial growth, promote blood vessel and tissue regeneration and accelerate wound healing. In the embodiment of the application, the mole percentage of XO in the bioactive glass is 15% -40%. B is₂O₃The molar percentage in the bioactive glass may in particular be, but not limited to, 15%, 20%, 30% or 40%. In some embodiments of the present application, XO comprises CaO, MgO, SrO, and CuO, wherein CaO is present in the bioactive glass in a molar percentage of 15% to 25%; the molar percentage of MgO in the bioactive glass is 5-10%; the mol percentage of SrO in the bioactive glass is 3% -8%; the mol percentage of CuO in the bioactive glass is 0.1-2%. When CaO, MgO, SrO and CuO are simultaneously included in the XO, the composite hydrogel is in Ca²⁺、Mg^{2 +}、Sr²⁺And Cu²⁺Under the synergistic effect of the components, the composite hydrogel can better realize the functions of promoting blood coagulation, promoting angiogenesis, regulating inflammation and resisting bacteria. In some embodiments of the present application, the molar percentages of CaO, MgO, SrO, and CuO in the bioactive glass are 22%, 8%, 6%, and 0.5%, respectively, and at the molar ratio, the synergistic effect between the elements can be very good. In some embodiments of the present application, the bioactive glass further comprises P₂O₅，P₂O₅The mole percentage in the bioactive glass is less than or equal to 4%. A certain content of P₂O₅Can regulate the degradation performance and bioactivity of the bioactive glass, and meanwhile, P is also an element required by metabolism of a human body, and can promote the repair of wounds.

In some embodiments of the present application, the bioactive glass is grafted with methacryloxy groups (MA) that are attached to the Bioactive Glass (BG) by siloxane bonds. In the embodiment of the application, the methacryloxy group is grafted to the bioactive glass to obtain the methacryloxy bioactive glass (BG-MA), and the methacryloxy bioactive glass (BG-MA) and the methacryloxy silk fibroin (SF-MA) are jointly crosslinked by grafting the methacryloxy bioactive glass to ensure that the formed composite hydrogel component is uniform and has good mechanical strength.

In the embodiment of the present application, the bioactive glass has a particle size of 0.5 μm to 5 μm. The particle size of the bioactive glass may specifically be, but not limited to, 0.5 μm, 1 μm, 3 μm or 5 μm. The activity of the bioactive glass can be weakened due to too large particle size, the antibacterial and anti-inflammatory performance is reduced, the activity of the bioactive glass can be too high due to too small particle size, the biocompatibility is reduced, the wound surface is stimulated, and the recovery of the wound surface is not facilitated.

In the embodiment of the application, the mass percentage of the bioactive glass in the antibacterial and anti-inflammatory composite hydrogel precursor solution is 1-6%. The mass percentage of the bioactive glass in the antibacterial and anti-inflammatory composite hydrogel precursor solution may be, but is not limited to, 1%, 3%, 5% or 6%. The water absorption of the composite hydrogel can be adjusted by a certain content of bioactive glass, so that the composite hydrogel can absorb seepage of a wound surface and has certain structural strength and is not easy to deform. In the embodiment of the application, the mass ratio of the methacryl oxidized silk fibroin to the bioactive glass is 2-5. The mass ratio of methacryloxy oxidized silk fibroin to bioactive glass may specifically but not exclusively be 2, 3, 4 or 5. The mass ratio of the methacryl oxidized silk fibroin to the bioactive glass is controlled, so that the composite hydrogel formed by the precursor liquid has good lubricating property and certain anti-inflammatory and antibacterial capabilities, and the wound surface is quickly healed.

In an embodiment of the present application, the photoinitiator comprises one or more of lithium phenyl-2, 4, 6-trimethylbenzoylphosphate, 1- [ 4- (2-hydroxyethoxy) -phenyl ] -2-hydroxy-2-methyl-1-propan-1-one, 2' -azo [ 2-methyl-N- (2-hydroxyethyl) propanamide ]. The photoinitiator can catalyze methacryloyl oxidation silk fibroin (SF-MA) to generate crosslinking under the action of ultraviolet light, so that the antibacterial and anti-inflammatory composite hydrogel precursor liquid forms the antibacterial and anti-inflammatory composite hydrogel. In the embodiment of the application, the mass percentage of the photoinitiator in the antibacterial and anti-inflammatory composite hydrogel precursor solution is 0.1-0.4%. The mass percentage of the photoinitiator in the antibacterial and anti-inflammatory composite hydrogel precursor solution may be, but is not limited to, 0.1%, 0.2%, 0.3% or 0.4%. The content of the photoinitiator can be controlled to ensure that the precursor liquid can realize sufficient and rapid crosslinking, and the influence of the photoinitiator on human bodies is small. In the embodiment of the application, the mass percentage of water in the antibacterial and anti-inflammatory composite hydrogel precursor solution is 64-88%.

In some embodiments, the antibacterial and anti-inflammatory composite hydrogel precursor solution further comprises one or more of an anti-inflammatory drug, an antibacterial drug, and a growth factor drug. When an anti-inflammatory drug, an antibacterial drug or a growth factor drug is added into the precursor solution, the formed composite hydrogel can continuously convey the drugs to the wound, so that a lasting treatment effect is achieved, and the repair of the wound surface is facilitated.

The antibacterial and anti-inflammatory composite hydrogel precursor solution can be crosslinked and cured under ultraviolet irradiation to form the antibacterial and anti-inflammatory composite hydrogel. In the embodiment, the composite hydrogel has a tensile strength of 20Kpa to 40 Kpa. The composite hydrogel has high mechanical strength, is not easy to break, can effectively protect the wound surface, and shields dust and bacteria; the composite hydrogel has good adhesion to the wound surface and is not easy to fall off, and in addition, the composite hydrogel can isolate and inhibit the growth of germs, promote the regeneration of wound surface tissues and blood vessels, inhibit inflammatory reaction and accelerate the healing of the wound surface.

The application also provides a preparation method of the antibacterial and anti-inflammatory composite hydrogel precursor liquid, which comprises the following steps: mixing the methacryl oxidized silk fibroin, the bioactive glass, the photoinitiator and water to obtain the antibacterial and anti-inflammatory composite hydrogel precursor solution. In some embodiments of the present application, the preparation method of the antibacterial and anti-inflammatory composite hydrogel precursor solution specifically comprises: dissolving methacryloyl oxidized silk fibroin (SF-MA) in water, adding a photoinitiator and bioactive glass to obtain a mixed solution, and ultrasonically dispersing the mixed solution at the power of 50-200W for 10-60 min to obtain the antibacterial and anti-inflammatory composite hydrogel precursor solution, wherein the antibacterial and anti-inflammatory composite hydrogel precursor solution comprises the following components in percentage by mass: methacryloxy oxidized silk fibroin: 10% -30%; bioactive glass: 1% -6%; photoinitiator (2): 0.1% -0.4%; water: 64 to 88 percent.

In some embodiments of the present application, a method for preparing methacryl oxidized silk fibroin comprises: placing the silkworm cocoon in water at 70-100 ℃ to remove sericin, placing the silkworm cocoon in a lithium bromide solution, adding Glycidyl Methacrylate (GMA), reacting at 40-60 ℃ for 3-6 h, adding the reaction solution into a dialysis bag, dialyzing with deionized water for 4-7 days, and freeze-drying the reaction solution to obtain the methacryloyl oxidation silk fibroin (SF-MA). In the application, the lithium bromide solution can dissolve silk fibroin in the silkworm cocoons, and after the silkworm cocoons are dissolved in the lithium bromide solution, the mass percentage content of the silk fibroin in the mixed solution is 10% -30%. In the embodiment of the application, the volume of the added glycidyl methacrylate accounts for 4-8% of the volume of the mixed solution. In the embodiment of the application, the cut-off molecular weight of the dialysis bag is 10000Da-15000 Da. In some embodiments of the present application, the dialysis bag has a molecular weight cut-off of 12000Da to 14000 Da.

In some embodiments of the present application, a method of making Bioactive Glass (BG) comprises: dissolving tetraethoxysilane, boric acid and metal salt in a solution containing ethanol, water and nitric acid, stirring for 3-6 h at the temperature of 30-50 ℃, then sealing the reaction solution and putting the reaction solution into a thermostat for aging treatment for 3-7 days, wherein the temperature of the aging treatment is 50-70 ℃; and (3) placing the reaction liquid in a program temperature control furnace for heat treatment, cooling, and then ball-milling and sieving the product to obtain bioactive glass powder (BG). In the embodiment of the present application, the metal element in the metal salt includes one or more of Ca, Mg, Sr and Cu, and the metal salt may be a nitrate of the above metal element, or other soluble salts. In the embodiment of the application, in the solution containing ethanol, water and nitric acid, the volume ratio of ethanol to water is 1 (0.3-1), the volume ratio of ethanol to water can be controlled to promote the dissolution of each reactant, the reaction rate can be adjusted, and the reaction speed is slowed down and the preparation time is prolonged due to the overlarge volume ratio of ethanol to water. The concentration of the nitric acid is 0.1mol/L-0.5 mol/L. In some embodiments of the present application, the temperature conditions for the heat treatment are: heating to 500-700 ℃ at the heating rate of 1-5 ℃/min, keeping the temperature for 2-5 h, and removing the organic components in the product by heat treatment. In the embodiment of the application, the bioactive glass powder obtained by ball milling and sieving the product has the particle size of 0.5-5 μm.

In some embodiments of the present application, the method for preparing the methacryloxy bioactive glass (BG-MA) comprises the following steps: dispersing bioactive glass in ethanol, adding water, hydrochloric acid and 3- (methacryloyloxy) propyl trimethoxy silane to obtain a mixed solution, reacting at 40-60 ℃ for 4-6 h, centrifuging, repeatedly washing the precipitate with ethanol and water, and drying to obtain the methacryloyloxy bioactive glass (BG-MA). In the embodiment of the application, the mass percentage of the bioactive glass in the ethanol is 2-4%, the volume of the added water accounts for 4-10% of the volume of the mixed solution, the volume of the added 3- (methacryloyloxy) propyl trimethoxy silane accounts for 2-4% of the volume of the mixed solution, and the concentration of hydrochloric acid in the mixed solution is 0.05-0.5 mol/L.

The preparation method of the antibacterial and anti-inflammatory composite hydrogel precursor solution is simple to operate, reaction conditions are easy to control, and the preparation method is suitable for industrial production.

The application also provides a method for applying the antibacterial and anti-inflammatory composite hydrogel precursor liquid to wound repair, which comprises the following steps:

the antibacterial and anti-inflammatory composite hydrogel precursor solution is coated on a wound surface and is irradiated by ultraviolet light to form the antibacterial and anti-inflammatory composite hydrogel, and the antibacterial and anti-inflammatory composite hydrogel can cover and adhere to the wound surface, so that the effects of protecting the wound surface, isolating and inhibiting the growth of germs, promoting the regeneration of wound surface tissues and blood vessels and inhibiting inflammatory reaction are achieved. This applicationIn the embodiment, the usage amount of the antibacterial and anti-inflammatory composite hydrogel precursor liquid on the wound surface is 200 μ L/cm²-500μL/cm². In the embodiment of the application, the wavelength of the ultraviolet light irradiation is 315nm-400nm, and the time of the ultraviolet light irradiation is 10s-60 s. In the application, the antibacterial and anti-inflammatory composite hydrogel can be replaced according to the condition of a wound surface, and in some embodiments of the application, the replacement period of the antibacterial and anti-inflammatory composite hydrogel is 2 days to 4 days.

The application also provides a dressing, which is obtained by crosslinking and curing the antibacterial and anti-inflammatory composite hydrogel precursor liquid after being irradiated by ultraviolet light, and the dressing is covered and adhered to a wound surface, so that the wound surface can be protected, the growth of germs can be isolated and inhibited, the regeneration of wound surface tissues and blood vessels can be promoted, the inflammatory reaction can be inhibited, and the wound surface healing can be accelerated.

The antibacterial and anti-inflammatory composite hydrogel precursor solution has the characteristics of wide raw material sources, low cost, simple preparation method, rapidness and convenience in implementation and operation and remarkable application effect, and has a high application value in the repair of soft tissues such as skin and the like, particularly the repair of diabetic chronic wounds which are difficult to heal.

The following further describes embodiments of the present application in terms of a number of examples.

Example 1

An antibacterial and anti-inflammatory composite hydrogel precursor solution and a preparation method thereof comprise the following steps:

(1) preparation of methacryloyl oxidized fibroin protein (SF-MA)

Cutting silkworm cocoons, putting the cut silkworm cocoons into boiling water, boiling for 20min to remove sericin (repeating for three times), washing for 3 times by using water, drying, weighing 20g of silkworm cocoons with sericin removed, adding the silkworm cocoons into 100mL of lithium bromide solution with the concentration of 9.3mol/L, dissolving for 1h in constant-temperature water bath at 60 ℃, then adding 4mL of glycidyl methacrylate into the mixed solution, reacting for 3h, transferring the reaction solution into a dialysis bag (the molecular weight cutoff is 12000 and 14000Da), dialyzing for 5 days by using deionized water, and freezing and drying the reaction solution to obtain the methacryloyl oxidation fibroin (SF-MA).

(2) Preparation of Bioactive Glass (BG)

40mL of ethyl orthosilicateDissolving ester, 1.8g of boric acid, 4.0mL of triethyl phosphate and 21.6g of calcium nitrate in a solution of 45mL of ethanol, 15mL of deionized water and 1mL of nitric acid, placing the solution in a thermostatic water bath at 40 ℃ for magnetic stirring for 4 hours, then sealing the solution, placing the solution in a thermostat at 60 ℃ for aging treatment for 3 days, carrying out heat treatment in a temperature programmed furnace (raising the temperature to 600 ℃ at a speed of 2 ℃/min and keeping the temperature for 180min), cooling the solution, carrying out ball milling and sieving on the product to obtain Bioactive Glass (BG), wherein the composition of the obtained bioactive glass is (mole percentage): SiO 2₂：60％；B₂O₃：5％；P₂O₅：4％；CaO：31％。

(3) Preparation of antibacterial and anti-inflammatory composite hydrogel precursor liquid

Dissolving 1g of SF-MA in 5mL of deionized water to obtain an SF-MA aqueous solution, adding 0.01g of phenyl-2, 4, 6-trimethylbenzoyl lithium phosphate, and then ultrasonically dispersing 0.3g of bioactive glass in the solution to obtain the antibacterial and anti-inflammatory composite hydrogel precursor solution.

Example 2

An antibacterial and anti-inflammatory composite hydrogel precursor solution and a preparation method thereof comprise the following steps:

(1) preparation method of methacryl oxidized silk fibroin (SF-MA) in example 2 is the same as that of example 1

(2) Preparation of Bioactive Glass (BG)

Dissolving 40mL of ethyl orthosilicate, 1.9g of boric acid, 4.4mL of triethyl phosphate, 16.9g of calcium nitrate, 6.7g of magnesium nitrate, 4.1g of strontium nitrate and 0.4g of copper nitrate in a solution of 45mL of ethanol, 15mL of deionized water and 1mL of nitric acid, placing the solution in a thermostatic water bath at 40 ℃ for magnetic stirring for 4 hours, sealing the solution, placing the solution in a thermostatic water bath at 60 ℃ for aging treatment for 3 days, carrying out heat treatment in a temperature programmed control furnace (heating to 600 ℃ at a rate of 2 ℃/min for heat preservation for 180min), cooling, and then carrying out ball milling and sieving on a product to obtain Bioactive Glass (BG), wherein the composition of the obtained bioactive glass is as follows: SiO 2₂：55％；B₂O₃：5％；P₂O₅：4％；CaO：21.5％；MgO：8％；SrO：6％；CuO：0.5％。

(3) Preparation of antibacterial and anti-inflammatory composite hydrogel precursor liquid

Dissolving 1g of SF-MA in 5mL of deionized water to obtain an SF-MA aqueous solution, adding 0.01g of phenyl-2, 4, 6-trimethylbenzoyl lithium phosphate, and then ultrasonically dispersing 0.3g of bioactive glass in the solution to obtain the antibacterial and anti-inflammatory composite hydrogel precursor solution.

Example 3

An antibacterial and anti-inflammatory composite hydrogel precursor solution and a preparation method thereof comprise the following steps:

(1) preparation of methacryloylated oxidized silk fibroin (SF-MA) in example 3 was performed in the same manner as in example 1

(2) Preparation of methacryloxy bioactive glass (BG-MA)

Example 3 bioactive glass was prepared the same as in example 2 except that example 3 grafted methacryloxy groups on the surface of the bioactive glass as follows: dispersing 2g of bioactive glass into 100mL of ethanol, adding 4mL of deionized water, 50 mu L of hydrochloric acid and 4mL of 3- (methacryloyloxy) propyl trimethoxy silane, reacting in a constant-temperature water bath at 60 ℃ for 4 hours, repeatedly centrifuging and washing with ethanol and water, and drying to obtain the methacryloyloxy bioactive glass (BG-MA).

(3) Preparation of antibacterial and anti-inflammatory composite hydrogel precursor liquid

Dissolving 1g of SF-MA in 5mL of deionized water to obtain an SF-MA aqueous solution, adding 0.01g of phenyl-2, 4, 6-trimethylbenzoyl lithium phosphate, and then ultrasonically dispersing 0.08g of BG-MA in the solution to obtain the antibacterial and anti-inflammatory composite hydrogel precursor solution.

Irradiating the antibacterial and anti-inflammatory composite hydrogel precursor solution by adopting an ultraviolet flashlight with the wavelength of 365nm and the MW for 20s to obtain the antibacterial and anti-inflammatory composite hydrogel (SF-MA-BG).

Example 4

In example 4, methacryloyloxyfibroin (SF-MA) and methacryloyloxybioactive glass (BG-MA) were prepared in the same manner as in example 1, except that BG-MA was added in an amount of 0.25g in the preparation of the antibacterial and anti-inflammatory composite hydrogel precursor solution.

Irradiating the antibacterial and anti-inflammatory composite hydrogel precursor solution by adopting an ultraviolet flashlight with the wavelength of 365nm and the MW for 20s to obtain the antibacterial and anti-inflammatory composite hydrogel (SF-MA-BG).

Example 5

The preparation methods of methacryloyloxyfibroin (SF-MA) and methacryloyloxybioactive glass (BG-MA) in example 5 were the same as in example 1, except that BG-MA was added in an amount of 0.28g in the preparation of the antibacterial and anti-inflammatory composite hydrogel precursor solution.

Irradiating the antibacterial and anti-inflammatory composite hydrogel precursor solution by adopting an ultraviolet flashlight with the wavelength of 365nm and the MW for 20s to obtain the antibacterial and anti-inflammatory composite hydrogel (SF-MA-BG).

Comparative example 1

The difference between comparative example 1 and example 1 is that the hydrogel precursor solution does not contain bioactive glass, and the preparation method of the hydrogel precursor solution comprises the following steps: 1g of SF-MA was dissolved in 5mL of deionized water, and 0.01g of lithium phenyl-2, 4, 6-trimethylbenzoyl phosphate was added to obtain a hydrogel precursor.

And (3) irradiating the hydrogel precursor solution by adopting an ultraviolet flashlight with the wavelength of 365nm and the MW for 20s to obtain the hydrogel (SF-MA).

Effects of the embodiment

In order to verify the performance of the antibacterial and anti-inflammatory composite hydrogel precursor solution prepared by the method, an effect embodiment is also provided by the method.

1) The methacryloyl oxidized silk fibroin (SF-MA) prepared in example 1 is characterized by using nuclear magnetic resonance, please refer to fig. 1, fig. 1 is a nuclear magnetic resonance spectrum of the methacryloyl oxidized silk fibroin (SF-MA) prepared in example 1, SF-MA is methacryloyl oxidized silk fibroin and SF is silk fibroin in fig. 1, and it can be seen from the peak position of SF-MA at 5.3/5.6ppm that example 1 successfully modifies MA group in the molecular structure of SF.

2) The bioactive glass of example 2 is analyzed for particle size by a particle size analyzer, referring to fig. 2, fig. 2 is a particle size distribution diagram of the bioactive glass prepared in example 2, and as can be seen from fig. 2, the particle size distribution of the bioactive glass prepared in example 2 is between 500nm and 1.5 μm, and the particle size is concentrated at 1 μm.

3) The Bioactive Glass (BG) in example 2 is characterized by an X-ray diffractometer, referring to fig. 3, fig. 3 is an X-ray diffraction pattern of the bioactive glass prepared in example 3, and it can be seen from fig. 3 that the phase structure of the bioactive glass is amorphous.

4) The Silk Fibroin (SF), the methacryloxy-oxidized silk fibroin (SF-MA), the Bioactive Glass (BG), the methacryloxy-oxidized bioactive glass (BG-MA) and the antibacterial and anti-inflammatory composite hydrogel (SF-MA-BG) in example 3 were characterized by an infrared spectrometer. Referring to FIG. 4, FIG. 4 is an infrared characterization chart of Silk Fibroin (SF), methacryloyloxyfibroin (SF-MA), Bioactive Glass (BG), methacryloyloxybioactive glass (BG-MA), and antibacterial and anti-inflammatory composite hydrogel (SF-MA-BG) prepared in example 3, which is shown in FIG. 4 at 1636cm^-1、1516cm^-1、1232cm^-1And 674cm^-1The characteristic peaks indicate that a large number of amide groups are present in SF, SF-MA and SF-MA-BG, corresponding to the characteristics of amino acid polymers, and 1060cm^-1The peaks at (a) are due to the random conformation in SF. 1460cm^-1And 1050cm^-1The broad peaks in (A) are attributed to B-O-B and Si-O-Si vibrations in the bioactive glass, indicating the basic composition of the bioactive glass.

5) The antibacterial and anti-inflammatory composite hydrogel (SF-MA-BG) obtained in example 3 was characterized by a scanning electron microscope, and the specific test method included: after the antibacterial and anti-inflammatory composite hydrogel (SF-MA-BG) is freeze-dried to remove water, the surface morphology of the antibacterial and anti-inflammatory composite hydrogel is observed by a Scanning Electron Microscope (SEM), referring to fig. 5, fig. 5 is an electron microscope representation of the dried antibacterial and anti-inflammatory composite hydrogel provided in example 3. As can be seen from FIG. 5, the dried SF-MA-BG is a porous loose structure, and no obvious agglomeration phenomenon of bioglass particles is observed.

6) The antibacterial and anti-inflammatory composite hydrogel (SF-MA-BG) of examples 3 to 5 and the hydrogel (SF-MA) of comparative example 1 were subjected to swelling property tests, specifically: and (3) freeze-drying the hydrogel, measuring the dry weight of the hydrogel, then placing the hydrogel into deionized water, soaking the hydrogel for 24 hours, and measuring the wet weight, wherein the wet weight/dry weight ratio is the swelling coefficient of the hydrogel. Referring to fig. 6, fig. 6 is a swelling performance test chart of the antibacterial and anti-inflammatory composite hydrogels of examples 3 to 5 and the hydrogel (SF-MA) of comparative example 1, and it can be seen from fig. 6 that, compared to comparative example 1 without adding bioactive glass, after the bioactive glass is added to the precursor solution of the examples, the water absorption of the composite hydrogel obtained by curing the precursor solution is also reduced, so as to ensure that the wound surface is not too dry and the composite hydrogel is not deformed due to excessive water absorption.

7) The mechanical property test of the antibacterial and anti-inflammatory composite hydrogel (SF-MA-BG) of examples 3 to 5 and the hydrogel (SF-MA) of comparative example 1 was carried out, and the test method specifically was: the hydrogel was made to a size of 40X 20X 2mm (length X width X thickness). After the two ends are clamped, a sample is stretched at the speed of 5mm/min by using a universal mechanical testing machine, and the maximum stress in the stretching process is taken as the tensile strength of the sample. Referring to fig. 7, fig. 7 is a mechanical property test chart of the antibacterial and anti-inflammatory composite hydrogels of examples 3 to 5 and the hydrogel (SF-MA) of comparative example 1, and it can be seen from fig. 7 that as the content of bioactive glass in the precursor liquid is increased, the tensile strength of the composite hydrogel obtained after the precursor liquid is cured is increased, and when the content of bioactive glass reaches a certain degree, the tensile strength of the composite hydrogel starts to be reduced, which is because the network structure of the hydrogel is damaged when the content of glass is too high, resulting in the decrease of mechanical properties.

8) In order to verify the repairing effect of hydrogel formed by the antibacterial and anti-inflammatory composite hydrogel precursor liquid on the diabetic wound, a diabetic rat skin wound model is constructed, and the effect of the antibacterial and anti-inflammatory composite hydrogel composition on wound healing is researched, wherein the method comprises the following steps: a10-week-old rat is injected with 1% streptozotocin solution (dosage: 30mg/kg) by mass percentage after fasting for 12h, and a wound surface of 1.5cm multiplied by 1.5cm is formed on the back skin of the rat after continuously feeding for 2 weeks. Then, 500 mul of the antibacterial and anti-inflammatory composite hydrogel precursor liquid (example 4) is dripped to the wound surface, an ultraviolet flashlight (365nm, 1300MW) is used for irradiating for 20s, the antibacterial and anti-inflammatory composite hydrogel precursor liquid can form the antibacterial and anti-inflammatory composite hydrogel at the wound surface to cover the wound surface, and the repairing effect of the antibacterial and anti-inflammatory composite hydrogel on the diabetic wound surface is researched by observing the change of the size of the wound surface; the control group 1 was prepared by dropping the hydrogel precursor solution of comparative example 1 onto the wound surface and irradiating the wound surface with an ultraviolet flashlight to form hydrogel, wherein the wound surface was covered with gauze without using any hydrogel. Referring to fig. 8, fig. 8 is a graph illustrating wound healing effects of a control group 1, a control group 2 and example 4, and it can be seen from fig. 8 that the antibacterial and anti-inflammatory composite hydrogel provided by the present application can significantly promote healing of a diabetic wound compared to the control group 1 and the control group 2. Referring to fig. 9, fig. 9 is a graph of H & E staining results of the wound surfaces of the control group 1, the control group 2 and the example 4, as is apparent from fig. 9, when the rat wound surfaces are treated by the antibacterial and anti-inflammatory composite hydrogel of the present application, the rat wound surfaces are substantially flat on day 7 and completely recovered on day 14, while the rat wound surfaces of the control group are still in a ulcerated state on day 7, and the recovery time of the wound surfaces is slow.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.