Spinning solution, and electrospinning membrane and preparation method therefor

Abstract

A spinning solution, and an electrospinning membrane and a preparation method therefor. The spinning solution comprises 5-20 parts by weight of an in-vivo degradable amorphous polymer, 0.5-3 parts by weight of a water-soluble polymer, and 0.01-1.5 parts by weight of an antibacterial agent and a solvent. The weight-average molecular weight of the in-vivo degradable amorphous polymer is not less than 80 thousand. When an electrospinning membrane is formed by the spinning solution, the performance of the antibacterial agent can be well exerted, and the growth of bacteria is effectively inhibited, thereby achieving a better antibacterial effect of the electrospinning membrane.

Description

Spinning solution, electrospinning membrane and preparation method thereof

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to Chinese patent application number 2024100835118, filed with the Patent Office of China on January 19, 2024, entitled “Spinning solution, electrospinning membrane and preparation method thereof”, the entire contents of which are incorporated herein by reference. Technical Field

[0003] The present application relates to the technical field of electrospinning, and in particular to a spinning solution, an electrospinning membrane and a preparation method thereof. Background Art

[0004] In the prior art, electrospun membranes can be applied to damaged tissue to promote tissue repair. Because damaged tissue is often accompanied by inflammation, antibacterial ingredients are often added to electrospun membranes to inhibit bacterial growth.

[0005] However, in actual use, the inventors have found that the effect of antibacterial agents is usually not very good, resulting in more bacterial proliferation in damaged tissues and slower healing of damaged tissues.

[0006] Application Contents

[0007] In view of the shortcomings of the existing technology, the purpose of the embodiments of the present application includes providing a spinning solution, an electrospinning membrane and a preparation method thereof. After the spinning solution forms an electrospinning membrane, it effectively inhibits bacterial growth, making the electrospinning membrane have a better antibacterial effect.

[0008] In a first aspect, embodiments of the present application provide a spinning solution comprising, by weight, 5 to 20 parts by weight of a biodegradable amorphous polymer, 0.5 to 3 parts by weight of a water-soluble polymer, 0.01 to 1.5 parts by weight of an antimicrobial agent, and a solvent. The weight-average molecular weight of the biodegradable amorphous polymer is not less than 80,000.

[0009] In the above technical solution, the in vivo degradable amorphous polymer has excellent biocompatibility and degradability. Therefore, in the spinning solution, the content of the in vivo degradable amorphous polymer with a higher molecular weight is higher. It is used as the main component of the spinning solution so that the electrospinning membrane obtained later has better biocompatibility and degradability. The electrospinning membrane with this component as the main component is prone to shrinkage after covering or filling the damaged tissue, resulting in the antibacterial agent not being able to play a good role. The inventor explored the cause of the shrinkage and found that because the tissue damaged environment contains exudate, the in vivo degradable amorphous polymer will absorb water and swell, and the amorphous molecular chain will slip, causing the glass transition temperature of the material to drop to human body temperature (36°C to 38°C) or below, thereby causing it to shrink. In the present application, a small amount of water-soluble polymer is added to the spinning solution, and the exudate from the damaged tissue will preferentially combine with the water-soluble polymer to cause swelling, making it difficult for the exudate from the damaged area to combine with the amorphous polymer that can be degraded in the body and change its glass transition temperature. This can limit the shrinkage of the spinning membrane to a certain extent, so that the effect of the antibacterial agent can be well exerted, effectively inhibiting bacterial growth, and making the antibacterial effect of the electrospinning membrane better.

[0010] In some embodiments, the water-soluble polymer includes a synthetic water-soluble polymer, the weight average molecular weight of the synthetic water-soluble polymer is not less than 50,000, and the antibacterial agent is a low molecular weight antibacterial agent, the weight average molecular weight of the low molecular weight antibacterial agent is not higher than 10,000.

[0011] In the aforementioned technical solution, the antimicrobial agent has a low molecular weight and does not form a fibrous structure on the electrospun membrane. Instead, it typically exists in the form of particles or short strands. After the electrospun membrane shrinks, some of the antimicrobial agent is easily trapped within the shrinking membrane, resulting in poor antimicrobial performance. However, the addition of a small amount of a synthetic water-soluble polymer with a higher molecular weight can improve this problem. This polymer can form filaments together with the biodegradable amorphous polymer. These filaments can preferentially bind to exudate from damaged tissues and stabilize the fibrous structure, making the electrospun membrane less susceptible to shrinkage and enhancing the antimicrobial performance.

[0012] In some embodiments, the spinning solution further comprises 0.1 to 3 parts by weight of a bioactive substance.

[0013] In the above technical solution, the addition of bioactive substances can be combined with low-molecular antibacterial agents to enable the electrospun membrane to promote cell proliferation, effectively inhibit bacterial growth, and facilitate tissue repair and regeneration.

[0014] In some embodiments, the weight average molecular weight of the in vivo degradable amorphous polymer is 80,000 to 300,000; the weight average molecular weight of the bioactive substance is greater than 10,000, the weight average molecular weight of the synthetic water-soluble polymer is 50,000 to 500,000; and the weight average molecular weight of the low molecular weight antibacterial agent is 1,000 to 10,000.

[0015] In the above technical solution, the weight-average molecular weight of the bioactive substance and the synthetic water-soluble polymer is 50,000 to 500,000. The molecular weight of the bioactive substance and the synthetic water-soluble polymer is relatively high. During the electrospinning process, they can form fiber filaments simultaneously with the amorphous polymer that can be degraded in the body, which can make the performance of the bioactive substance better. When combined with low-molecular antibacterial agents, the antibacterial and proliferation effects of the electrospun membrane are better, which is beneficial to tissue repair and regeneration.

[0016] In some embodiments, the weight average molecular weight of the in vivo degradable amorphous polymer is 80,000 to 300,000; the weight average molecular weight of the bioactive substance is 1,000 to 10,000; the weight average molecular weight of the synthetic water-soluble polymer is 50,000 to 500,000; and the weight average molecular weight of the low molecular weight antibacterial agent is 1,000 to 10,000.

[0017] In the above technical solution, the combination of a high-molecular-weight, biodegradable amorphous polymer and a synthetic water-soluble polymer can impart excellent biocompatibility, biodegradability, and shrinkage resistance to the electrospun membrane. Furthermore, when combined with a small amount of a bioactive substance and an antimicrobial agent, both of which have a lower molecular weight, the properties of both can be fully utilized.

[0018] In some embodiments, the spinning solution comprises: 5 to 20 parts by weight of an in vivo degradable amorphous polymer, 0.1 to 2 parts by weight of a bioactive substance, 0.5 to 2 parts by weight of a synthetic water-soluble polymer, and 0.1 to 1.5 parts by weight of an antibacterial agent.

[0019] In the above technical solution, adding a small amount of synthetic water-soluble polymer can better inhibit the shrinkage of the electrospun membrane, so that after adding less bioactive substances and antibacterial agents, the functionality of the two can be well exerted to improve the comprehensive performance of the electrospun membrane.

[0020] In some embodiments, the weight portion X of the in vivo degradable amorphous polymer is greater than the sum of the weight portions Y of the bioactive substance, the synthetic water-soluble polymer, and the antimicrobial agent.

[0021] In the above technical solution, the in vivo degradable amorphous polymer is the main component of the spinning solution, which can make the biocompatibility and degradation effect of the electrospinning membrane better, and is combined with a synthetic water-soluble polymer. Even if the amount of antibacterial agent and bioactive substance added is very small, the performance of both can be well exerted.

[0022] In some embodiments, X>5×Y.

[0023] In some embodiments, the in vivo degradable amorphous polymer comprises at least one of racemic polylactic acid and poly(lactic-co-glycolic acid).

[0024] In the above technical solution, the above materials are selected to have good in vivo degradability and biocompatibility so as to serve as the main component of the electrospinning membrane.

[0025] In some embodiments, the bioactive substance includes at least one of collagen, growth factors, silk fibroin, zein, gelatin, and hyaluronic acid.

[0026] In the above technical solution, the selection of the above-mentioned bioactive substances has a good effect of promoting cell proliferation, so that the electrospun membrane can promote tissue repair and regeneration.

[0027] In some embodiments, the synthetic water-soluble polymer includes at least one of polyethylene oxide, polyethylene glycol, polyvinyl alcohol, polyacrylamide, and polyvinyl pyrrolidone.

[0028] In the above technical solution, adding a small amount of the above components can better improve the anti-shrinkage effect of the electrospun membrane, so that the performance of the antibacterial agent and the active substance can be better exerted.

[0029] In some embodiments, the antimicrobial agent comprises an inorganic antimicrobial agent and / or an organic antimicrobial agent.

[0030] In the above technical solutions, the molecular weight of the antibacterial agent is relatively small. If the electrospun membrane shrinks, the effect of the antibacterial agent is not easy to be exerted. Therefore, the antibacterial agent is combined with other components in the spinning solution to better exert the effect of the antibacterial agent.

[0031] In some embodiments, the organic antimicrobial agent comprises at least one of a guanidine antimicrobial agent, a β-lactam antimicrobial agent, and a quaternary ammonium salt antimicrobial agent.

[0032] In some embodiments, the guanidine antimicrobial agent includes at least one of polyhexamethyleneguanidine, polyhexamethylenebiguanide, polyaminopropylbiguanide, and dodecylguanidine acetate.

[0033] In some embodiments, the beta-lactam antibacterial agent comprises a penicillin and / or a cephalosporin.

[0034] In some embodiments, the quaternary ammonium salt antimicrobial agent includes at least one of alkyl trimethyl ammonium chloride, alkyl dimethyl benzyl ammonium chloride, and tetradecyl-2-methylpyridinium bromide.

[0035] In some embodiments, the solute mass percentage of the spinning solution is 5% to 30%.

[0036] In the above technical solution, the solute mass ratio of the spinning solution is within the above range, and it is easy to be stabilized into fiber filaments through electrospinning equipment, and the fibrous structure of the electrospinning membrane can simulate the extracellular matrix, thereby covering or filling the damaged tissue, which is beneficial to tissue repair and regeneration.

[0037] In a second aspect, the present application provides an electrospinning method, using the spinning solution provided in any one of the first aspects, and spraying it onto a receiving device under the conditions of a spinning voltage of 5kV to 30kV and a spinning solution flow rate of 0.2ml / h to 1ml / h.

[0038] In the above technical solution, the electrospun membrane obtained by the above spinning conditions is suitable for covering or filling damaged tissues, which is beneficial to promoting cell proliferation and can effectively inhibit bacterial growth, thereby accelerating tissue repair and regeneration.

[0039] In some embodiments, the temperature of the spinning environment is 20° C. to 35° C., and the humidity is 35% to 60%.

[0040] In the above technical solution, spinning is carried out under the temperature and humidity environment, which can make the spinning more stable and is conducive to forming a stable electrospinning membrane.

[0041] In some embodiments, the receiving device is a receiving roller, and the rotation speed of the receiving roller is 300 r / min to 3000 r / min.

[0042] In the above technical solution, the nanofibers of the electrospinning membrane can be distributed more evenly, which is beneficial to the volatilization of the solvent in the spinning solution during the spinning process.

[0043] In a third aspect, the present application provides an electrospinning membrane obtained by the electrospinning method provided in any one of the second aspects.

[0044] In the above technical solution, the electrospun membrane prepared by this method is suitable for covering or filling damaged tissues, which is beneficial to promoting cell proliferation and can effectively inhibit bacterial growth, thereby accelerating tissue repair and regeneration. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following is a brief introduction to the drawings required for use in the embodiments. It should be understood that the following drawings only show certain embodiments of the present application and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without creative work.

[0046] FIG1 is a graph showing the antibacterial performance of electrospun membranes provided in some examples and comparative examples of the present application. DETAILED DESCRIPTION

[0047] Electrospun membranes are fibrous films manufactured through electrospinning, exhibiting high strength and flexibility. To impart functionalities to electrospun membranes, substances with corresponding functionalities are typically added to the electrospinning solution for electrospinning. For example, to impart antibacterial properties to an electrospun membrane, an antimicrobial agent is typically added to the solution to inhibit bacterial growth.

[0048] Electrospun membranes with antibacterial properties are usually covered or filled in damaged tissues (for example, the tissue can be skin, subcutaneous fascia, muscle, tendon sheath, ligament, peripheral nerves and blood vessels, etc.). Therefore, if the electrospun membrane has good in vivo absorption or metabolism properties and good biocompatibility, the performance of the electrospun membrane can be made better. Therefore, an amorphous polymer that can be degraded in the body (for example, polylactic acid-glycolic acid copolymer) can be used as the base material of the electrospun membrane. After the base material forms the electrospun membrane, it will cover or fill the damaged tissue for a period of time, and shrinkage problems will usually occur, resulting in the antibacterial agent in the electrospun membrane not being able to perform well.

[0049] The inventors studied the reasons for the shrinkage of electrospun membranes and found that under dry conditions, the glass transition temperature of polylactic acid-co-glycolic acid copolymer is about 48°C (its deformation temperature is higher than the ambient temperature and human body temperature). However, due to the presence of a certain amount of exudate in the damaged tissue, the polylactic acid-co-glycolic acid copolymer matrix component in the electrospun membrane easily absorbs water and swells, causing the amorphous molecular chains to slip. After absorbing water for a period of time, the glass transition temperature of the material is reduced, making the glass transition temperature consistent with or below the human body temperature (36°C to 38°C), thereby causing it to shrink. Therefore, based on the problem that the in vivo degradable amorphous polymer is prone to shrinkage after covering or filling the damaged tissue for a period of time, the spinning solution, electrospinning method and electrospinning membrane are improved as follows.

[0050] Spinning solution

[0051] The present application provides a spinning solution comprising, by weight, 5 to 20 parts by weight of a biodegradable amorphous polymer, 0.5 to 3 parts by weight of a water-soluble polymer, 0.01 to 1.5 parts by weight of an antimicrobial agent, and a solvent. The weight-average molecular weight of the biodegradable amorphous polymer is not less than 80,000.

[0052] Degradable includes in vitro biodegradability (for example, land degradation, environmental degradation, etc.) and in vivo biodegradability (absorbable or metabolizable in vivo). In vitro biodegradability does not mean that it has good biocompatibility (referring to safety and non-toxicity to the human body, low or no rejection reaction), while in vivo biodegradability usually has good biocompatibility, so that the electrospun membrane formed can cover or fill the damaged tissue. An in vivo degradable amorphous polymer means that the polymer is both a high molecular weight polymer that can be absorbed or metabolized in the body and an amorphous polymer. There will be a certain swelling effect in media such as water or ethanol, which will cause the molecular chains to slide relative to each other. Therefore, the electrospun membrane formed by the in vivo degradable amorphous polymer after spinning will cause the membrane to shrink after covering or filling the damaged tissue for a long time. The present application is an improvement based on the shrinkage characteristics of the polymer. The weight average molecular weight of the in vivo degradable amorphous polymer is not less than 80,000. In the electrospinning membrane formed by the spinning solution, the component that forms the fiber filaments is mainly the in vivo degradable amorphous polymer, which is the main body of the electrospinning membrane.

[0053] Water-soluble polymers refer to biological water-soluble polymers and / or synthetic water-soluble polymers. Biological water-soluble polymers refer to polymers whose raw materials are derived from biological sources (e.g., bioactive substances). Synthetic water-soluble polymers refer to polymers whose raw materials are artificially synthesized and do not exist in natural biological materials. The water-soluble polymers in this application can be of biological or artificial origin, and can also be added simultaneously, and this application does not limit them.

[0054] In such a spinning solution, the in vivo degradable amorphous polymer has excellent biocompatibility and degradability. Therefore, in the spinning solution, the content of the in vivo degradable amorphous polymer with a higher molecular weight is higher. It is used as the main component of the spinning solution so that the electrospinning membrane obtained later has better biocompatibility and degradability. The electrospinning membrane with this component as the main component is prone to shrinkage after covering or filling the damaged tissue, resulting in the antibacterial agent not being able to play a good role. The inventor explored the cause of the shrinkage and found that because the tissue damaged environment contains exudate, the in vivo degradable amorphous polymer will absorb water and swell, and the amorphous molecular chain will slip, causing the glass transition temperature of the material to drop to human body temperature (36°C to 38°C) or below, thereby causing it to shrink. In the present application, a small amount of water-soluble polymer is added to the spinning solution, and the exudate from the damaged tissue will preferentially combine with the water-soluble polymer to cause swelling, making it difficult for the exudate from the damaged tissue to combine with the amorphous polymer that can be degraded in the body and change its glass transition temperature. This can limit the shrinkage of the electrospun membrane to a certain extent, thereby enabling the antibacterial agent to exert its effect well, effectively inhibiting bacterial growth, and making the antibacterial effect of the electrospun membrane better.

[0055] In some embodiments, the water-soluble polymer comprises a synthetic water-soluble polymer having a weight-average molecular weight of no less than 50,000, and the antimicrobial agent is a low-molecular-weight antimicrobial agent having a weight-average molecular weight of no more than 10,000. The synthetic water-soluble polymer has a relatively high molecular weight and can be combined with the in vivo degradable amorphous polymer to form fibers. The antimicrobial agent has a relatively low molecular weight and is essentially incapable of forming fibers, typically being in the form of particles or short strands.

[0056] Due to the low molecular weight of the antimicrobial agent, it does not form a fibrous structure on the electrospun membrane. Instead, it typically exists in the form of particles or short strands. After the electrospun membrane shrinks, some of the antimicrobial agent is easily trapped inside the shrinking electrospun membrane, resulting in poor antimicrobial performance. However, the addition of a small amount of a synthetic water-soluble polymer with a higher molecular weight can improve this problem. This polymer can form filaments together with the biodegradable amorphous polymer. These filaments can preferentially bind to the exudate from damaged tissue and also stabilize the fibrous structure, making the electrospun membrane less susceptible to shrinkage and enhancing the antimicrobial performance.

[0057] In some embodiments, the spinning solution further comprises 0.1 to 3 parts by weight of a bioactive substance. The addition of the bioactive substance can be combined with a low-molecular-weight antimicrobial agent to enable the electrospun membrane to promote cell proliferation, effectively inhibit bacterial growth, and facilitate tissue repair and regeneration.

[0058] The bioactive substance may or may not be water-soluble. In some embodiments, the bioactive substance is a water-soluble bioactive substance, and the spinning solution comprises 5 to 20 parts by weight of an in vivo degradable amorphous polymer, 0.5 to 3 parts by weight of the water-soluble bioactive substance, 0.01 to 1.5 parts by weight of an antimicrobial agent, and a solvent. The weight-average molecular weight of the in vivo degradable amorphous polymer is not less than 80,000.

[0059] In some embodiments, the bioactive substance has a relatively high weight-average molecular weight. The spinning solution comprises, by weight, 5-20 parts by weight of a biodegradable amorphous polymer, 0.1-3 parts by weight of the bioactive substance, 0.5-3 parts by weight of a synthetic water-soluble polymer, 0.01-1.5 parts by weight of an antimicrobial agent, and a solvent. The weight-average molecular weight of the biodegradable amorphous polymer is 80,000 to 300,000; the weight-average molecular weight of the bioactive substance is greater than 10,000; the weight-average molecular weight of the synthetic water-soluble polymer is 50,000 to 500,000; and the weight-average molecular weight of the low-molecular-weight antimicrobial agent is 1,000 to 10,000.

[0060] In the spinning solution, the biodegradable amorphous polymer and the bioactive substance with a high weight-average molecular weight simultaneously form filaments. The organic antimicrobial agent has a smaller molecular weight and cannot form filaments. Instead, it forms short linear or granular substances on the electrospun membrane. When the electrospun membrane is covered or filled with damaged tissue, the environment at the damaged tissue contains exudate. If the covering or filling time is long, the biodegradable amorphous polymer will absorb water and swell, and the amorphous molecular chains will slip, causing the glass transition temperature of the biodegradable amorphous polymer to drop to human body temperature (36°C to 38°C) or below, causing the electrospun membrane to shrink. After shrinkage, the short linear or granular organic antimicrobial agent is partially wrapped in the shrunken membrane, and its performance cannot be fully exerted, making it difficult to inhibit bacterial growth. However, the bioactive substance forms filaments and shrinks along with the electrospun membrane, allowing the performance of the bioactive substance to be still well exerted. Although the bioactive substance can promote cell proliferation, it also promotes bacterial division. In addition, the performance of the antimicrobial agent itself is not fully exerted, and the bacterial load of the exudate is increased. If the amount of antibacterial agent is increased, the biocompatibility of the electrospun membrane will be poor. Therefore, the present application adds a synthetic water-soluble polymer with a higher weight-average molecular weight, which can be simultaneously filamented with the amorphous polymer that can be degraded in the body and the bioactive substance during the electrospinning process. The exudate from the damaged tissue preferentially combines with the synthetic water-soluble polymer to cause swelling, and the exudate from the damaged tissue is not easy to combine with the amorphous polymer that can be degraded in the body to swell. The synthetic water-soluble polymer also plays a role in stabilizing the fibrous structure, improving the shrinkage of the electrospun membrane, and making the performance of the antibacterial agent better. At the same time, the bioactive substance can promote cell proliferation, making the electrospun membrane conducive to tissue repair and regeneration.

[0061] As an example, the weight average molecular weight of the in vivo degradable amorphous polymer is 80,000, 100,000, 120,000, 140,000, 160,000, 180,000, 200,000, 220,000, 240,000, 260,000, 280,000 or 300,000; the weight average molecular weight of the synthetic water-soluble polymer is 50,000, 100,000, 150,000, 200,000, 250,000, 300,000, 350,000, 400,000, 450,000 or 500,000; the weight average molecular weight of the bioactive substance is 10,000, 50,000, 100,000, 150,000, 200,000, 250,000, 300,000, 350,000, 400,000, 450,000 or 500,000; the weight average molecular weight of the organic antibacterial agent is 1,000, 2,000, 3,000, 4,000, 5,000, 6,000, 7,000, 8,000, 9,000 or 10,000.

[0062] Optionally, the weight average molecular weight of the in vivo degradable amorphous polymer is 100,000 to 300,000; the weight average molecular weight of the bioactive substance and the synthetic water-soluble polymer are both 50,000 to 300,000. The weight average molecular weights of the three fiber-forming polymers are relatively uniform, so as to form a more uniform electrospun membrane, which can further improve the problem of incomplete performance of the antibacterial agent due to shrinkage of the electrospun membrane, thereby making the performance of the antibacterial agent and the bioactive substance better. It should be noted that the weight average molecular weights of the bioactive substance and the synthetic water-soluble polymer are both 50,000 to 300,000, which does not mean that the weight average molecular weights of the two are the same. The weight average molecular weights of the two can be the same or different, and this application does not limit it.

[0063] In other embodiments, the weight-average molecular weight of the bioactive substance is relatively low. The spinning solution comprises, by weight, 5-20 parts of a biodegradable amorphous polymer, 0.1-3 parts of the bioactive substance, 0.5-3 parts of a synthetic water-soluble polymer, 0.01-1.5 parts of an antimicrobial agent, and a solvent. The weight-average molecular weight of the biodegradable amorphous polymer is 80,000-300,000; the weight-average molecular weight of the bioactive substance is 1,000-10,000; the weight-average molecular weight of the synthetic water-soluble polymer is 50,000-500,000; and the weight-average molecular weight of the low-molecular-weight antimicrobial agent is 1,000-10,000.

[0064] In the spinning solution, the body-degradable amorphous polymer can be formed into fiber filaments, and the molecular weight of the organic antimicrobial agent and the bioactive substance is too small to form fiber filaments. Instead, short-line or granular substances will be formed on the electrospinning membrane. When the electrospinning membrane is covered or filled on the damaged tissue, since the environment of the damaged tissue contains exudate, a long covering or filling time will cause the body-degradable amorphous polymer to absorb water and swell, and the amorphous molecular chain will slip, resulting in the glass transition temperature of the body-degradable amorphous polymer being reduced to human body temperature (36°C ~ 38°C) or below, causing the electrospinning membrane to shrink. After shrinkage, the short-line or granular organic antimicrobial agent and bioactive substance are partially wrapped in the shrunken membrane, and their performance cannot be well exerted, and the effect of inhibiting bacterial growth and promoting cell proliferation is poor. If the amount of antibacterial agents and bioactive substances is increased, the biocompatibility of the electrospun membrane will be poor. Therefore, the present application adds a synthetic water-soluble polymer with a higher weight-average molecular weight, which can be simultaneously formed into fibers with the amorphous polymer that can be degraded in the body and the bioactive substance during the electrospinning process. The exudate from the damaged tissue preferentially combines with the synthetic water-soluble polymer to cause swelling, and the exudate from the damaged tissue is not easy to combine with the amorphous polymer that can be degraded in the body to swell. The synthetic water-soluble polymer also plays a role in stabilizing the filamentous structure, improving the shrinkage of the electrospun membrane, and making the performance of the antibacterial agent and the bioactive substance better, thereby inhibiting the growth of bacteria and promoting cell proliferation, making the electrospun membrane conducive to tissue repair and regeneration.

[0065] As an example, the weight average molecular weight of the in vivo degradable amorphous polymer is 80,000, 100,000, 120,000, 140,000, 160,000, 180,000, 200,000, 220,000, 240,000, 260,000, 280,000 or 300,000; the weight average molecular weight of the synthetic water-soluble polymer is 50,000, 100,000, 150,000, 200,000, 250,000, 300,000, 350,000, 400,000, 450,000 or 500,000; the weight average molecular weight of the bioactive substance is 1,000, 3,000, 5,000, 7,000, 9,000 or 10,000; the weight average molecular weight of the organic antibacterial agent is 1,000, 2,000, 3,000, 4,000, 5,000, 6,000, 7,000, 8,000, 9,000 or 10,000.

[0066] Optionally, the weight-average molecular weight of the in vivo degradable amorphous polymer is 100,000 to 300,000; the weight-average molecular weight of the synthetic water-soluble polymer is 100,000 to 300,000; and the weight-average molecular weight of the bioactive substance is 1,000 to 10,000. The weight-average molecular weights of the two fiber-forming polymers are relatively uniform, so as to form a more uniform electrospun membrane. This can further improve the problem of incomplete performance of the antibacterial agent due to shrinkage of the electrospun membrane, thereby improving the performance of both the antibacterial agent and the bioactive substance. It should be noted that the weight-average molecular weights of the in vivo degradable amorphous polymer and the synthetic water-soluble polymer are both 100,000 to 300,000, which does not mean that the weight-average molecular weights of the two are the same. The weight-average molecular weights of the two can be the same or different, and this application does not limit this.

[0067] In other embodiments, the weight-average molecular weight of the in vivo degradable amorphous polymer may be 320,000, 340,000, 360,000, 380,000, 400,000, 420,000, 440,000, 460,000, 480,000, or 500,000; the weight-average molecular weight of the synthetic water-soluble polymer may be 520,000, 540,000, 560,000, 580,000, or 600,000; the weight-average molecular weight of the organic antimicrobial agent may be 500, 600, 700, 800, or 900; and the weight-average molecular weight of the bioactive substance may be 550,000 or 600,000.

[0068] As an example, in terms of weight parts, the weight part X of the in vivo degradable amorphous polymer in the spinning solution can be 5 parts, 8 parts, 10 parts, 12 parts, 15 parts, 18 parts or 20 parts; the weight part Y1 of the bioactive substance can be 0.1 parts, 0.5 parts, 1 parts, 1.5 parts, 2 parts, 2.5 parts or 3 parts; the weight part Y2 of the water-soluble polymer can be 0.5 parts, 1 parts, 1.5 parts, 2 parts, 2.5 parts or 3 parts; the weight part Y3 of the antibacterial agent can be 0.1 parts, 0.5 parts, 1 parts, 1.5 parts, 2 parts, 2.5 parts, 3 parts, 3.5 parts, 4 parts, 4.5 parts or 5 parts.

[0069] Optionally, the spinning solution includes 5 to 20 parts by weight of a biodegradable amorphous polymer, 0.1 to 2 parts by weight of a bioactive substance, 0.5 to 2 parts by weight of a synthetic water-soluble polymer, and 0.1 to 1.5 parts by weight of an antimicrobial agent. Adding a small amount of the synthetic water-soluble polymer effectively inhibits shrinkage of the electrospun membrane, thereby effectively utilizing the properties of the bioactive substance and antimicrobial agent even with the addition of smaller amounts, thereby improving the overall performance of the electrospun membrane.

[0070] Furthermore, the weight of the biodegradable amorphous polymer (X) is greater than the sum of the weights of the bioactive substance, the synthetic water-soluble polymer, and the antimicrobial agent (Y) (Y = Y1 + Y2 + Y3). The biodegradable amorphous polymer, as the primary component of the spinning solution, enhances the biocompatibility and degradation performance of the electrospun membrane. Furthermore, when combined with the synthetic water-soluble polymer, the performance of the antimicrobial agent and bioactive substance can be fully utilized even with minimal additions.

[0071] Furthermore, X>5×Y. This can not only make the effects of the bioactive substances and antibacterial agents fully exerted, but also make the electrospun membrane have good biocompatibility and degradability.

[0072] In some embodiments, the in vivo degradable amorphous polymer comprises at least one of racemic polylactic acid and poly(lactic-co-glycolic acid). This material is selected for its good in vivo degradability and biocompatibility, allowing it to be used as the main component of the electrospun membrane for repairing damaged tissues.

[0073] In the present application, the specific composition of the in vivo degradable amorphous polymer is not limited, as long as it is a biodegradable and biocompatible amorphous polymer.

[0074] In some embodiments, the bioactive substance includes at least one of collagen, growth factors, silk fibroin, zein, gelatin, and hyaluronic acid. The selection of the above-mentioned bioactive substances has a good effect of promoting cell proliferation, so that the electrospun membrane can promote tissue repair and regeneration.

[0075] Bioactive substances can be either water-soluble or water-insoluble. In some embodiments, the bioactive substance is water-soluble (for example, the water-soluble bioactive substance is one or more of growth factors, gelatin, and hyaluronic acid). It can be added to the spinning solution as both a bioactive substance and a water-soluble polymer. Adding the water-soluble bioactive substance also adds the water-soluble polymer, simplifying the composition of the spinning solution. In other embodiments, even if a water-soluble bioactive substance is added to the spinning solution, a synthetic water-soluble polymer can also be added to the spinning solution.

[0076] In some embodiments, among the bioactive substances, collagen, silk fibroin, zein, gelatin, and hyaluronic acid are bioactive substances with a weight average molecular weight greater than or equal to 10,000. Among the bioactive substances, growth factors and enzymatically cleaved oligomeric sodium hyaluronate are bioactive substances with a weight average molecular weight less than 10,000.

[0077] In the present application, the specific composition of the bioactive substance is not limited, as long as it is a bioactive substance that can promote cell proliferation.

[0078] In some embodiments, the synthetic water-soluble polymer includes at least one of polyethylene oxide, polyethylene glycol, polyvinyl alcohol, polyacrylamide, and polyvinyl pyrrolidone. These synthetic water-soluble polymers, when added in small amounts, can effectively enhance the anti-shrinkage properties of the electrospun membrane, thereby effectively enhancing the performance of the antimicrobial agent and bioactive substances.

[0079] In the present application, the composition of the synthetic water-soluble polymer is not limited, as long as it is a polymer that can improve the shrinkage performance of the electrospun membrane whose main component is an amorphous polymer that can be degraded in vivo.

[0080] In some embodiments, low-molecular-weight antimicrobial agents include inorganic and / or organic antimicrobial agents. These antimicrobial agents have relatively low molecular weights, and if the electrospun membrane shrinks, the antimicrobial agent's effectiveness is less pronounced. Therefore, the antimicrobial agent is combined with other components in the spinning solution to maximize its effectiveness.

[0081] The inorganic antibacterial agent includes at least one of silver sulfate, zinc oxide, copper oxide, ammonium dihydrogen phosphate and lithium carbonate.

[0082] The organic antimicrobial agent includes at least one of a guanidine antimicrobial agent, a β-lactam antimicrobial agent and a quaternary ammonium salt antimicrobial agent.

[0083] In some embodiments, the guanidine antimicrobial agent includes at least one of polyhexamethyleneguanidine, polyhexamethylenebiguanide, polyaminopropylbiguanide and dodecylguanidine acetate. β-lactam antimicrobial agents include penicillin and / or cephalosporin. Quaternary ammonium salt antimicrobial agents include at least one of alkyltrimethylammonium chloride, alkyldimethylbenzylammonium chloride and tetradecyl-2-methylpyridinium bromide. These antimicrobial agents are organic antimicrobial agents. After electrospinning, they are more likely to form short lines. The shrinkage of the electrospinning membrane has a relatively small effect on the performance of the antimicrobial agent (compared to inorganic antimicrobial agents). Therefore, when combined with other components in the spinning solution (for example, synthetic water-soluble polymers), the effect of the organic antimicrobial agent can be better exerted.

[0084] In the present application, the specific composition of the antibacterial agent is not limited. As long as the antibacterial agent has antibacterial properties and can be used in the spinning solution, it is within the protection scope of the present application.

[0085] In some embodiments, the solvent includes one or more of hexafluoroisopropanol, chloroform, dichloromethane, trifluoroacetic acid, dimethyl sulfoxide, tetrahydrofuran, dimethylformamide, acetic acid, water, methanol, ethanol, and isopropanol. These solvents can dissolve other components in the spinning solution better, which is conducive to the subsequent formation of an electrospinning film by electrostatic spinning. If the solvent is only water, it cannot dissolve all the components in the spinning solution, but it can be combined with other solvents to dissolve other components in the spinning solution. The solvent selection in this application is one or more, which is related to the selection of other components in the spinning solution. Those skilled in the art can make adjustments according to actual conditions.

[0086] In the present application, the composition of the solvent is not limited. As long as the solvent can dissolve other components in the spinning solution, it is within the protection scope of the present application.

[0087] In some embodiments, the solute mass percentage of the spinning solution is 5% to 30%, wherein the solute is the substance other than the solvent in the spinning solution, and the solute mass percentage is solute / (solute+solvent)×100%.

[0088] The solute mass ratio of the spinning solution is within the above range, and it is easy to form fiber filaments through the electrospinning equipment. In addition, the fiber structure of the electrospinning membrane can simulate the extracellular matrix, which is beneficial to the repair and regeneration of tissue when covering or filling the damaged tissue.

[0089] As an example, the solute mass percentage of the spinning solution is 5%, 10%, 15%, 20%, 25% or 30%. Optionally, the solute mass percentage of the spinning solution is 10% to 20%.

[0090] Method for preparing spinning solution

[0091] The present application provides a method for preparing a spinning solution, wherein the other components in the aforementioned spinning solution are dispersed in a solvent. The components in the spinning solution are the aforementioned components, and the components of the spinning solution are not described in detail here. The preparation method will be mainly described.

[0092] In some embodiments, a biodegradable amorphous polymer is first dispersed in a solvent to form a mixed solution 1, and then a bioactive substance, a synthetic water-soluble polymer, and an antimicrobial agent are added and dispersed in the mixed solution 1 to form a spinning solution. The addition of a relatively high amount of the biodegradable amorphous polymer allows it to be dissolved and dispersed in the solvent first, followed by the dissolution and dispersion of the other components. This can improve the dissolution and dispersion of the spinning solution, resulting in a more uniform spinning solution.

[0093] In other embodiments, each component may be dispersed in a solvent separately and then mixed together to form a spinning solution. Alternatively, other components may be dispersed in a solvent first, and then the in vivo degradable amorphous polymer may be dispersed.

[0094] Electrospinning method

[0095] The present application provides an electrospinning method using the aforementioned spinning solution for electrospinning, comprising: spraying the spinning solution onto a receiving device at a spinning voltage of 5kV to 30kV and a spinning solution flow rate of 0.2ml / h to 1ml / h. The electrospun membrane obtained under these spinning conditions is suitable for covering or filling damaged tissues, promoting cell proliferation and effectively inhibiting bacterial growth, thereby accelerating tissue repair and regeneration.

[0096] For example, the spinning voltage is 5 kV, 10 kV, 15 kV, 20 kV, 25 kV, or 30 kV; the spinning solution flow rate is 0.2 ml / h, 0.4 ml / h, 0.6 ml / h, 0.8 ml / h, or 1 ml / h. Alternatively, the spinning voltage is 5 kV to 20 kV; and the spinning solution flow rate is 0.4 ml / h to 0.8 ml / h.

[0097] In some embodiments, the spinning environment has a temperature of 20° C. to 35° C. and a humidity of 35% to 60%. Spinning under such a temperature and humidity environment can make the spinning more stable and facilitate the formation of a stable electrospun membrane.

[0098] Illustratively, the temperature of the spinning environment is 20°C, 22°C, 25°C, 28°C, 30°C, 32°C or 35°C; and the humidity is 35%, 40%, 45%, 50%, 55% or 60%.

[0099] In some embodiments, the receiving device is a receiving roller, and the speed of the receiving roller is 300 rpm to 3000 rpm, which can make the nanofibers of the electrospinning membrane more evenly distributed and facilitate the volatilization of the solvent in the spinning solution during the spinning process.

[0100] As an example, the speed of the receiving roller is 300r / min, 500r / min, 800r / min, 1000r / min, 1200r / min, 1500r / min, 1800r / min, 2000r / min, 2200r / min, 2500r / min, 2800r / min or 3000r / min. Optionally, the speed of the receiving roller is 500r / min to 2000r / min.

[0101] electrospun membrane

[0102] The present application provides an electrospun membrane, obtained by subjecting the above-mentioned spinning solution to the aforementioned electrospinning method. The electrospun membrane produced by this method has good biocompatibility and can improve shrinkage of the electrospun membrane. It is suitable for covering or filling damaged tissues, promoting cell proliferation and effectively inhibiting bacterial growth, thereby accelerating tissue repair and regeneration.

[0103] In other embodiments, the spinning solution can also be formed on the receiving device by coating to form a membrane material. The spinning solution in this application is not limited to forming a membrane by electrospinning, and other spinning methods or coating methods are also within the scope of protection of this application.

[0104] To make the purpose, technical solutions and advantages of the examples of the present application clearer, the technical solutions in the examples of the present application will be described clearly and completely below. Where specific conditions are not specified in the examples, conventional conditions or conditions recommended by the manufacturer are used. Where the manufacturer of the reagents or instruments is not specified, they are all conventional products that can be purchased commercially.

[0105] Example 1

[0106] A spinning solution comprises, by weight, 20 g of poly(lactic-co-glycolic acid) (PLGA, weight-average molecular weight of 100,000), 0.4 g of hydrolyzed collagen (Col, collagen, weight-average molecular weight of 200,000), 1 g of polyethylene oxide (PEO, polyethylene oxide, weight-average molecular weight of 400,000), 0.3 g of polyhexamethylene biguanide hydrochloride (PHMB, weight-average molecular weight of 1500) and 78.3 g of hexafluoroisopropanol (HFIP, hexafluoroisopropanol).

[0107] The method for preparing the spinning solution comprises:

[0108] Step 1: Using a precision electronic balance, 20 g of poly(lactic-co-glycolic acid) particles were weighed and added to 82.3 g of hexafluoroisopropanol. A magnetic stirrer was added and stirred at room temperature until clear and transparent to obtain a PLGA solution.

[0109] Step 2: Using a precision electronic balance, weigh 0.4 g of hydrolyzed collagen freeze-dried tablets, 1 g of polyethylene oxide powder, and 0.3 g of polyhexamethylene biguanide hydrochloride powder into the PLGA solution obtained in step 1, and stir at room temperature until clear and transparent to obtain a PHMB / Col / PEO / PLGA spinning solution;

[0110] Step 3: Place the PHMB / Col / PEO / PLGA spinning solution into a syringe. Attach the syringe to the electrospinning machine's syringe pump. Install a needle connected to the positive terminal of a high-voltage power supply. Connect the receiving roller to the negative terminal of the high-voltage power supply. Set the spinning parameters: spinning voltage 10 kV, spinning solution flow rate 0.5 ml / h, receiving roller speed 800 rpm, temperature 28°C, and humidity 58%.

[0111] Step 4: Dry the nanofiber material prepared by the above spinning at room temperature for 12 hours to obtain a PHMB / Col / PLGA (containing PEO) nanofiber membrane.

[0112] Example 2

[0113] Example 2 is basically the same as Example 1, except that the polylactic acid-glycolic acid copolymer in Example 1 is replaced by racemic polylactic acid (PDLLA, Poly (D, L-lactide), D, L-polylactide, weight average molecular weight of 100,000) powder in Example 2; and the solvent hexafluoroisopropanol in Example 1 is replaced by tetrahydrofuran in Example 2.

[0114] Example 3

[0115] Example 3 is basically the same as Example 1, except that no polyethylene oxide is added.

[0116] Example 4

[0117] Example 4 is basically the same as Example 1, except that the polyethylene oxide powder in Example 1 is replaced by polyvinyl alcohol (PVA, Polyvinyl alcohol, weight average molecular weight of 80,000) powder in Example 4.

[0118] Example 5

[0119] Example 5 is basically the same as Example 1, except that the polyethylene oxide powder in Example 1 is replaced by polyvinyl pyrrolidone (PVP, Polyvinyl pyrrolidone, weight average molecular weight of 60,000) powder in Example 5.

[0120] Example 6

[0121] Example 6 is basically the same as Example 1, except that the collagen freeze-dried sheet in Example 1 is replaced with 2g of gelatin (Gel, Gelatin, weight-average molecular weight of 80,000) particles in Example 6, and the added amount of polyhexamethylene biguanide hydrochloride is 0.1g.

[0122] Example 7

[0123] Example 7 is basically the same as Example 1, except that the collagen in Example 1 is replaced with 0.5 g of fibroblast growth factor (FGF, Fibroblast Growth factor, weight-average molecular weight of 6,000) freeze-dried powder in Example 7, and the added amount of polylactic acid-glycolic acid copolymer is 10 g.

[0124] Example 8

[0125] Example 8 is basically the same as Example 1, except that the polyhexamethylene biguanide hydrochloride in Example 1 is replaced by 0.01 g of amoxicillin (molecular weight 365.4) in Example 8, and the added amount of polylactic acid-glycolic acid copolymer is 10 g.

[0126] Example 9

[0127] Example 9 is basically the same as Example 1, except that the polyhexamethylene biguanide hydrochloride in Example 1 is replaced by 0.01 g of silver sulfate (molecular weight of 169.87) in Example 9, and the amount of polylactic acid-glycolic acid copolymer added is 18 g.

[0128] Example 10 to Example 11

[0129] Examples 10 and 11 are basically the same as Example 1, except that the addition amounts of the components are different. See Table 1 for details.

[0130] Example 12

[0131] A spinning solution comprises, by weight, 10 g of poly(lactic-co-glycolic acid) (PLGA, weight-average molecular weight of 100,000), 0.4 g of collagen (Col, weight-average molecular weight of 200,000), 3 g of polyethylene oxide (PEO, weight-average molecular weight of 400,000), 0.3 g of polyhexamethylene biguanide hydrochloride (PHMB, weight-average molecular weight of 1500) and 86.3 g of hexafluoroisopropanol (HFIP, hexafluoroisopropanol).

[0132] The method for preparing the spinning solution comprises:

[0133] Step 1: Using a precision electronic balance, 10 g of poly(lactic-co-glycolic acid) particles were weighed and added to 86.3 g of hexafluoroisopropanol. A magnetic stirrer was added and stirred at room temperature until clear and transparent to obtain a PLGA solution.

[0134] Step 2: Using a precision electronic balance, 0.4 g of collagen, 3 g of polyethylene oxide powder, and 0.3 g of polyhexamethylene biguanide hydrochloride powder were weighed into the PLGA solution obtained in step 1, and stirred at room temperature until clear and transparent to obtain a PHMB / Col / PEO / PLGA spinning solution;

[0135] Step 3: Place the PHMB / Col / PEO / PLGA spinning solution into a syringe. Attach the syringe to the electrospinning machine's syringe pump. Install a needle connected to the positive terminal of a high-voltage power supply. Connect the receiving roller to the negative terminal of the high-voltage power supply. Set the spinning parameters: spinning voltage 10 kV, spinning solution flow rate 0.5 ml / h, receiving roller speed 800 rpm, temperature 28°C, and humidity 58%.

[0136] Step 4: Dry the nanofiber material prepared by the above spinning at room temperature for 12 hours to obtain a PHMB / Col / PLGA (containing PEO) nanofiber membrane.

[0137] Comparative Example 1

[0138] A spinning solution comprises, by weight, 16g of poly(lactic-co-glycolic acid) (PLGA, with a weight-average molecular weight of 100,000) and 84g of hexafluoroisopropanol (HFIP, Hexafluoroisopropanol).

[0139] The method for preparing the spinning solution comprises:

[0140] Step 1: Using a precision electronic balance, 16 g of poly(lactic-co-glycolic acid) particles were weighed into 84 g of hexafluoroisopropanol, a magnetic stirrer was added, and the mixture was stirred at room temperature until clear and transparent to obtain a PLGA solution;

[0141] Step 2: Place the PLGA solution in a syringe. Attach the syringe to the electrospinning machine's syringe pump. Install a needle connected to the positive terminal of a high-voltage power supply. Connect the receiving roller to the negative terminal of the high-voltage power supply. Set the spinning parameters: spinning voltage 10 kV, dope flow rate 0.5 ml / h, receiving roller speed 800 rpm, temperature 28°C, and humidity 58%.

[0142] Step 3: Dry the nanofiber material prepared by the above spinning at room temperature for 12 hours to obtain a PLGA nanofiber membrane.

[0143] Comparative Example 2

[0144] A spinning solution comprises, by weight, 16 g of poly(lactic-co-glycolic acid) (PLGA, with a weight-average molecular weight of 100,000), 0.5 g of polyhexamethylene biguanide hydrochloride (PHMB, with a weight-average molecular weight of 1500), and 83.5 g of hexafluoroisopropanol (HFIP).

[0145] The method for preparing the spinning solution comprises:

[0146] Step 1: Using a precision electronic balance, 16 g of poly(lactic-co-glycolic acid) particles were weighed into 83.5 g of hexafluoroisopropanol, a magnetic stirrer was added, and the mixture was stirred at room temperature until clear and transparent to obtain a PLGA solution;

[0147] Step 2: Using a precision electronic balance, weigh 0.5 g of polyhexamethylene biguanide hydrochloride powder into the PLGA solution obtained in step 1, and stir at room temperature until clear and transparent to obtain a PHMB / PLGA spinning solution;

[0148] Step 3: Place the PHMB / PLGA spinning solution into a syringe. Attach the syringe to the electrospinning machine's syringe pump. Install a needle connected to the positive terminal of a high-voltage power supply. Connect the receiving roller to the negative terminal of the high-voltage power supply. Set the spinning parameters: spinning voltage 10 kV, dope flow rate 0.5 ml / h, receiving roller speed 800 rpm, temperature 28°C, and humidity 58%.

[0149] Step 4: Dry the nanofiber material prepared by the above spinning at room temperature for 12 hours to obtain the PHMB / PLGA nanofiber membrane.

[0150] Comparative Example 3

[0151] A spinning solution comprises, by weight, 16g of polyvinyl butyral (PVB, Polyvinlbutaral, with a weight-average molecular weight of 100,000) and 84g of an ethanol aqueous solution.

[0152] The method for preparing the spinning solution comprises:

[0153] Step 1: Using a precision electronic balance, 16 g of polyvinyl butyral ester particles were weighed and added to 84 g of ethanol aqueous solution. A magnetic stirrer was added and stirred at room temperature until the solution became clear and transparent to obtain a PVB solution.

[0154] Step 2: Place the PVB solution in a syringe. Attach the syringe to the electrospinning machine's syringe pump. Install a needle connected to the positive terminal of a high-voltage power supply. Connect the receiving roller to the negative terminal of the high-voltage power supply. Set the spinning parameters: spinning voltage 10 kV, dope flow rate 0.5 ml / h, receiving roller speed 800 rpm, temperature 28°C, and humidity 58%.

[0155] Step 3: Dry the nanofiber material prepared by the above spinning at room temperature for 12 hours to obtain a PVB nanofiber membrane.

[0156] Experimental Example 1

[0157] The performance of the electrospinning membranes provided by Examples 1 to 12 and Comparative Examples 1 to 3 were tested, and the results are shown in Table 1.

[0158] (1) Antibacterial performance: Tested according to GB / T 20944.1-2007, ATCC8739 Escherichia coli (E. coli) was used as the test strain. Antibacterial performance was evaluated by the size of the inhibition zone formed after the sample was added. The inhibition zone indicates that no bacteria grow in the area and the growth of surrounding bacteria is inhibited. The larger the inhibition zone, the better the antibacterial performance.

[0159] The specific method is as follows: All samples are first cut into a circle with a diameter of 3.55 cm and sterilized under ultraviolet light for 2 hours. Sterile nutrient agar is poured into a sterile petri dish and cooled under ultraviolet sterilization for later use. Then 150 μl of bacterial solution (concentration of 10 5 The sterilized samples were placed in the center of the agar plates and placed in a biological incubator at 37±1℃ for 24h.

[0160] Determination of the inhibition zone: First, observe whether there is an area without bacterial growth around the electrospun membrane: if there is, it means that the bacteria stop growing when they reach the outer ring of the area, which proves that there is a significant antibacterial effect, and the inhibition area is the area of ​​the outer ring; if not, continue to use a point light source to irradiate the electrospun membrane area to check whether there are colonies in the membrane-covered area. If so, it means that the membrane has no antibacterial effect and the inhibition area is 0; if not, it means that the bacteria have not entered the film-covered area to grow, which proves that the membrane has a certain antibacterial effect and can prevent bacteria from entering the dressing and growing inside.

[0161] The antibacterial performance diagram of some samples is shown in Figure 1, wherein the inner ring in the culture dish in Figure 1 represents the electrospun membrane, and the gray area around the inner ring represents the inhibition zone. As can be seen from Figure 1, compared with Example 1 and Example 10, the PLGA electrospun membranes provided by Comparative Examples 1 and 2 have obvious shrinkage, and no inhibition zone is formed, and do not have antibacterial properties. Therefore, it can be seen from Comparative Example 1 that the PLGA electrospun membrane provided by Comparative Example 1 is easy to shrink and does not have antibacterial properties; it can be seen from Comparative Example 2 that the PHMB antibacterial agent is added to the PLGA electrospun membrane, and the obtained electrospun membrane still has obvious shrinkage, and the performance of the antibacterial agent is not able to be exerted, and there is still no antibacterial effect; it can be seen from Examples 1 and 10 that the addition of synthetic water-soluble polymer polyethylene oxide to the electrospun membrane can significantly improve the shrinkage of the electrospun membrane, and the antibacterial effect of the electrospun membrane is effectively improved.

[0162] (2) Cell activity assay

[0163] The mouse fibroblast L929 cell line was used for the cell viability experiment. The experimental groups were set up with each test sample, and the control group did not contain any sample. First, the experimental groups were placed in 12-well tissue culture plates and sterilized with UV for 2 hours. Then, L929 cells (1×10 5 / mL) were inoculated into 12-well tissue culture plates (12 parallel experiments were performed for each sample and the test results were analyzed as shown in Table 1). The samples were all placed in a 5% CO2 incubator at 37°C and incubated. The incubation time was 72h. After the incubation period, the absorbance value of each group at 570nm was measured using an MTT method using a microplate reader. By calculating the absorbance ratio, the cell proliferation rate of each experimental group can be obtained. The higher the cell proliferation rate, the better the cell activity, that is, the stronger the ability to promote tissue repair; if the cell proliferation rate is less than 100%, it means that the sample is cytotoxic.

[0164] Table 1 Composition of spinning solution and properties of electrospinning membrane

[0165] As can be seen from Table 1, in Comparative Example 3, PVB will cause certain cytotoxicity and affect cell growth activity. Comparison between Comparative Example 1 and Comparative Example 2 shows that PLGA has good biocompatibility, is not toxic to cells, and the electrospun nanofiber structure also has a certain promoting effect on cell activity. However, when PLGA is used as the main spinning solution, even if the antibacterial agent PHMB is added to the spinning solution, the obtained electrospun membrane basically has no antibacterial effect. In Examples 1 to 12 of the present application, when an amorphous polymer that can be degraded in vivo is used as the main spinning solution, adding a small amount of water-soluble polymer and antibacterial agent can increase the antibacterial area of ​​the electrospun membrane and make the performance of the antibacterial agent better.

[0166] Comparison between Example 1 and Example 2 shows that the in vivo degradable amorphous polymer is selected as PLGA or PDLLA, and is combined with a synthetic water-soluble polymer (PEO), a bioactive substance (Col, a natural water-soluble polymer) and an antibacterial agent (PHMB). This can improve the performance of the antibacterial agent and the bioactive substance, inhibit bacterial growth (larger inhibition area), and promote cell proliferation (high cell proliferation rate), which is beneficial to tissue repair and regeneration.

[0167] From the comparison between Example 1 and Example 3, it can be seen that when the in vivo degradable amorphous polymer PLGA is used as the main body of the spinning solution, compared with Example 3 in which only bioactive substances (Col, natural water-soluble polymers) are added, Example 1 simultaneously adds synthetic water-soluble polymers (PEO) and bioactive substances (Col, natural water-soluble polymers), which can further enhance the performance of the antibacterial agent and the bioactive substance, further inhibit the growth of bacteria (larger inhibition area), and further promote cell proliferation, which is beneficial to tissue repair and regeneration.

[0168] From the comparison of Example 1 and Examples 4 to 9, it can be seen that the synthetic water-soluble polymer selected as PEO, PVA or PVP, the bioactive substance selected as Col, gelatin or FGF, and the antibacterial agent selected as PHMB, amoxicillin or silver sulfate can effectively inhibit the growth of bacteria and promote cell proliferation, which is beneficial to tissue repair and regeneration.

[0169] The embodiments described above are part of the embodiments of the present application, rather than all of the embodiments. The detailed description of the embodiments of the present application is not intended to limit the scope of the present application for protection, but merely represents selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

Claims

1. A spinning solution, characterized in that, By weight, it includes: 5 to 20 parts by weight of a biodegradable amorphous polymer, 0.5 to 3 parts by weight of a water-soluble polymer, 0.01 to 1.5 parts by weight of an antibacterial agent, and a solvent; Among them, the weight-average molecular weight of the biodegradable amorphous polymer is not less than 80,000.

2. The spinning solution according to claim 1, characterized in that, The water-soluble polymer includes a synthetic water-soluble polymer, the weight-average molecular weight of the synthetic water-soluble polymer is not less than 50,000, the antibacterial agent is a low-molecular antibacterial agent, and the weight-average molecular weight of the low-molecular antibacterial agent is not higher than 10,000.

3. The spinning solution according to claim 2, characterized in that, The spinning solution further includes 0.1 to 3 parts by weight of a bioactive substance.

4. The spinning solution according to claim 3, characterized in that, The weight-average molecular weight of the biodegradable amorphous polymer is 80,000 to 300,000; the weight-average molecular weight of the bioactive substance is greater than 10,000, the weight-average molecular weight of the synthetic water-soluble polymer is 50,000 to 500,000; the weight-average molecular weight of the low-molecular antibacterial agent is 1,000 to 10,000.

5. The spinning solution according to claim 3, characterized in that, The weight-average molecular weight of the biodegradable amorphous polymer is 80,000 to 300,000; the weight-average molecular weight of the bioactive substance is 1,000 to 10,000; the weight-average molecular weight of the synthetic water-soluble polymer is 50,000 to 500,000; the weight-average molecular weight of the low-molecular antibacterial agent is 1,000 to 10,000.

6. The spinning solution according to claim 3, characterized in that, The spinning solution includes: 5 to 20 parts by weight of the biodegradable amorphous polymer, 0.1 to 2 parts by weight of the bioactive substance, 0.5 to 2 parts by weight of the synthetic water-soluble polymer, and 0.1 to 1.5 parts by weight of the antibacterial agent; Optionally, the weight part X of the biodegradable amorphous polymer is greater than the sum Y of the weight parts of the bioactive substance, the synthetic water-soluble polymer, and the antibacterial agent; Optionally, X > 5×Y.

7. The spinning solution according to any one of claims 3 to 6, characterized in that, The spinning solution satisfies at least one of the following conditions: a) The biodegradable amorphous polymer includes at least one of racemic polylactic acid and poly(lactic-co-glycolic acid); b) The bioactive substance includes at least one of collagen, growth factor, silk fibroin, zein, gelatin, and hyaluronic acid; c) The synthetic water-soluble polymer includes at least one of polyethylene oxide, polyethylene glycol, polyvinyl alcohol, polyacrylamide, and polyvinylpyrrolidone; d) The antibacterial agent includes an organic antibacterial agent and / or an inorganic antibacterial agent.

8. The spinning solution according to claim 7, characterized in that, The organic antibacterial agent includes at least one of guanidine antibacterial agents, β-lactam antibacterial agents, and quaternary ammonium salt antibacterial agents.

9. The spinning solution according to claim 8, characterized in that, The organic antibacterial agent satisfies at least one of the following conditions: e) The guanidine antibacterial agent includes at least one of polyhexamethylene guanidine, polyhexamethylene biguanide, polyaminopropyl biguanide, and dodecylguanidine acetate; f) The β-lactam antibacterial agent includes penicillin and / or cephalosporin; g) The quaternary ammonium salt antibacterial agent includes at least one of alkyltrimethylammonium chloride, alkyldimethylbenzylammonium chloride, and 2-methylpyridinium bromide tetradecyl.

10. The spinning solution according to any one of claims 1 to 6, characterized in that, The mass ratio of the solute in the spinning solution is 5% to 30%.

11. A method for preparing an electrospun membrane, characterized in that, Using the spinning solution according to any one of claims 1 to 10, it is sprayed onto a receiving device under the conditions that the spinning voltage is 5 kV to 30 kV and the spinning solution flow rate is 0.2 ml / h to 1 ml / h.

12. The electrospinning method according to claim 11, wherein The temperature of the spinning environment is 20°C to 35°C, and the humidity is 35% to 60%. Optionally, the receiving device is a receiving roller, and the rotational speed of the receiving roller is 300 r / min to 3000 r / min.

13. An electrospun membrane, characterized in that, Obtained by the method for preparing an electrospun membrane according to claim 11 or 12.

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