A polyelectrolyte composite nanofiber membrane prepared by coaxial double-row electrospinning process and preparation method thereof

Abstract

The present invention discloses a method for preparing a polyelectrolyte composite nanofiber membrane using a coaxial double-row electrospinning process. The steps are as follows: S1, dissolving polyvinyl alcohol, quaternized chitosan, and modified alginate in water respectively to obtain solutions a, b, and c; S2, using these raw materials through coaxial double-row electrospinning, solution a is the nanofiber core layer, and b and c are the shell layers; S3, high-voltage electrospinning is used to prepare a coaxial nanofiber composite hemostatic membrane. The invention has significant advantages, simple preparation process, high porosity of nanofibers, mimicking the structure of natural fibrin and extracellular matrix, and good biocompatibility. After encountering blood, electrostatic action occurs to form a dense network with excellent hemostatic properties. The coaxial co-spinning process makes the material have stronger electrostatic action, high adhesion to biological tissues, can be peeled off on demand with physiological saline and is harmless, can be used for wound closure and hemostasis, and has broad application prospects in the field of wound dressings.

Description

Technical Field

[0001] The present invention belongs to the field of medical polymer materials, and specifically relates to a polyelectrolyte composite nanofiber membrane prepared by a coaxial double-row electrospinning process and a preparation method thereof. The nanofiber is made of pure natural raw materials, has good biocompatibility, and has efficient hemostatic properties. Background Art

[0002] Acute hemorrhage is one of the leading causes of death worldwide each year. Practical pre-hospital care solutions for acute hemorrhage control are urgently needed, especially in war, disaster, and accident situations, which are characterized by high clinical demand and significant market potential. Rapid and effective application of hemostatic materials in the early stages of rebleeding can prolong rescue time, thereby reducing the high mortality rate caused by excessive blood loss. However, rapid and safe control of bleeding from severe wounds or cuts remains challenging for most hemostatic materials. Hemostatic materials must be designed and developed to achieve rapid and complete hemostasis for both non-compressible (visceral organs) and compressible (femoral artery) hemorrhages, both of which are associated with life-threatening bleeding and contribute to equivalent consequences.

[0003] So far, various hemostatic products have been reported, such as gelatin sponge ® )、Chitosan gauze (HemCon ® ), hemostatic powder (Celox™), inorganic particle embedded gauze (QuikClot ® A variety of hemostatic products, such as styptic powders, have been developed and are currently in clinical use. However, each has its own drawbacks: sponges exhibit good shape memory due to their inherent interconnected macroporous structure, but suffer from poor mechanical strength; porous zeolites can cause thermal damage to surrounding tissues and, like hemostatic powders, are prone to distal thrombosis; and gauze must be frequently replaced and can easily adhere to wounds, causing secondary damage.

[0004] Quaternized chitosan and alginate are both natural high-molecular-weight polysaccharides with low cost, abundant sources, and excellent biocompatibility and biodegradability, making them promising applications in the field of regenerative hemostasis. Nanofibers, due to their large surface area, high porosity, breathability, and extracellular matrix-like structural characteristics, can support the adhesion and migration of fibroblasts during wound healing after hemostasis, promoting tissue regeneration. Summary of the Invention

[0005] In order to overcome the technical problems of poor mechanical strength of the above-mentioned hemostatic materials, easy to cause distal thrombosis and easy to cause secondary damage, and to prepare an efficient hemostatic nanofiber auxiliary material that does not require any toxic organic solvents, has good biocompatibility and can interact with groups on the wound, the present invention provides a cross-linking agent, a degradable and efficient hemostatic sponge used for non-pressable wounds. The purpose of the present invention is to provide a polyelectrolyte composite nanofiber membrane prepared by a coaxial double-row electrospinning process and a preparation method thereof. The present invention ingeniously adopts the method of coaxial double-row electrospinning to prepare a composite nanofiber membrane hemostatic auxiliary material using two natural high molecular polymers with polyelectrolyte anions and cations with opposite charges. The corresponding components are dissolved by a simple aqueous solution as a solvent, without the need for glutaraldehyde as a cross-linking agent, or UV treatment and other steps, to obtain flexible nanofibers. Because two polyelectrolytes with opposite charges are introduced as the shell layer of the nanofiber, when the nanofiber encounters blood, electrostatic interaction can occur between the two fibers, thereby forming a dense network structure. At the same time, the material and the groups on the tissue can also interact, thereby achieving an adhesion effect. The above two effects can ensure that the nanofiber can immediately and effectively physically block and adhere to the wound during the hemostasis process.

[0006] The present invention is achieved through the following technical solutions:

[0007] S1. dissolving polyvinyl alcohol in deionized water to obtain solution a;

[0008] S2, dissolving the polyelectrolyte cation in an aqueous acetic acid solution to obtain solution b;

[0009] S3, dissolving the polyelectrolyte anion in the aqueous solution to obtain solution c;

[0010] S4. Using the solution a obtained in step S1 as the core layer of the nanofiber, and the solution b obtained in step S2 and the solution c obtained in step S3 as the shell layer of the nanofiber, electrospinning is performed to obtain polyelectrolyte composite nanofibers.

[0011] As a preferred embodiment, in step S1, the polyvinyl alcohol needs to have a certain molecular weight sufficient to be stretched by the electric field to form continuous fibers. In step S1, the degree of polymerization of the polyvinyl alcohol is 4,000 to 100,000; the polyvinyl alcohol solution is dissolved in deionized water at 90°C, and the reaction time is 3 hours.

[0012] As a preferred embodiment, the polyelectrolyte anion is one of carboxymethyl chitosan, alginate, hyaluronic acid, and carboxymethyl cellulose, and the alginate is modified alginate; the polyelectrolyte cation is one of chitosan quaternary ammonium salt and chitosan, and the chitosan quaternary ammonium salt is quaternized chitosan.

[0013] As a preferred embodiment, the added amounts of polyvinyl alcohol, quaternized chitosan, modified alginate, and acetic acid are 5-10%, 0.5-2.5%, 0.5-3%, and 50-90%, respectively; the added amounts are calculated as a percentage by mass of the total mixture of polyvinyl alcohol, quaternized chitosan, modified alginate, and acetic acid.

[0014] As a preferred embodiment, the degree of substitution of the quaternized chitosan is 80% to 98%, the dissolution time is 8 h, and the reaction temperature is room temperature.

[0015] As a preferred solution, in step S3, the dissolution temperature of the modified alginate is 0-2°C, and the dissolution time is 8 hours.

[0016] As a preferred solution, the spinning temperature of electrospinning is 37°C, the humidity is 20-30%, the distance between the plates is 20 cm, the voltage is 21 kV, the flow rate of the liquid in the double-row spinning is 0.09 µL / min, and the spinning time is 12 h.

[0017] The polyelectrolyte composite nanofiber hemostatic auxiliary material is obtained by the preparation method of the present invention.

[0018] The present invention adopts the above technical solution, and the polyelectrolyte composite nanofiber membrane hemostatic auxiliary material can generate electrostatic interaction when it comes into contact with water to form a dense network structure, and interact with groups on the tissue, thereby completing immediate physical blocking.

[0019] Compared with the prior art, the method of the present invention has the following beneficial effects:

[0020] The nanofiber hemostatic auxiliary material of the present invention does not use any toxic reagents, does not have any harmful substance residues, is safe and harmless to the human body, and has no antigenicity.

[0021] The nanofiber membrane hemostatic auxiliary material shown in the present invention has blood thixotropy, can quickly form a dense network structure, and adhere to tissues to physically seal wounds. At the same time, the material has certain antibacterial properties, can effectively inhibit the formation of bacteria, and promote wound healing.

[0022] The raw materials of the nanofiber membrane hemostatic auxiliary material of the present invention are low in cost, easy to obtain, simple in method, and environmentally friendly.

[0023] The nanofiber membrane hemostatic auxiliary material of the present invention directly acts on bleeding wounds, is simple and convenient to use, and is easy to store, transport and carry. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a diagram showing the preparation of the polyelectrolyte nanofabric prepared in the present invention.

[0025] Figure 2This is a physical picture of the polyelectrolyte nanofiber hemostatic auxiliary material prepared by the present invention.

[0026] Figure 3 The in vitro coagulation indexes of the polyvinyl alcohol nanofabric prepared in Comparative Example 1 of the present invention, the quaternized chitosan nanofabric of Comparative Example 2, the modified alginate nanofabric of Comparative Example 3, the polyelectrolyte nanofabric prepared in Example 4, the medical gauze, and the combat gauze are 71%, 58%, 56%, 8%, 85%, and 79% respectively; the in vitro coagulation index of the polyvinyl alcohol nanofabric prepared in Comparative Example 1 is 85%, indicating that it does not have a good red blood cell capture ability, that is, it cannot quickly enrich coagulation factors to form a thrombus and achieve a hemostatic effect. The numerical values ​​of the polyelectrolyte composite nanofabric prepared by the coaxial double-row electrospinning process show that the nanofabric composed of nanofibers with opposite charges can not only rely on its own positive charge to enrich red blood cells and platelets, but also undergo electrostatic interaction after encountering blood, thereby forming a dense network structure, so that the polyelectrolyte nanofabric prepared in Example 3 exhibits excellent in vitro coagulation ability.

[0027] Figure 4 The hemostatic effects of the polyvinyl alcohol nanofabric prepared in Comparative Example 1, the quaternized chitosan nanofabric prepared in Comparative Example 2, the modified alginate nanofabric prepared in Comparative Example 3, and the polyelectrolyte nanofabric hemostatic supplement, medical gauze, and combat gauze prepared in Example 4 of the present invention in a rat femoral artery injury model were investigated. The polyvinyl alcohol nanofabric prepared in Comparative Example 1, the quaternized chitosan nanofabric prepared in Comparative Example 2, and the modified alginate nanofabric prepared in Comparative Example 3 all failed to effectively seal the wound and achieve a hemostatic effect. However, the polyelectrolyte composite nanofabric prepared by the coaxial double-row electrospinning process demonstrated that, because the nanofabric composed of oppositely charged nanofibers not only undergoes electrostatic interaction upon contact with blood, but also chemically reacts with tissue, achieving adhesion. Therefore, the polyelectrolyte nanofabric prepared in Example 3 exhibited excellent hemostatic effects in rat femoral arteries.

[0028] Figure 5 The hemostatic effects of the polyvinyl alcohol nanofabric prepared in comparative example 1 of the present invention, the quaternized chitosan nanofabric prepared in comparative example 2, the modified alginate nanofabric prepared in comparative example 3, the polyelectrolyte nanofabric hemostatic adjuvant prepared in Example 4, the medical gauze, and the combat gauze on the rat liver puncture model wound were studied. DETAILED DESCRIPTION

[0029] The present invention will be described in detail below with reference to specific embodiments. The following embodiments will help those skilled in the art to further understand the present invention, but are not intended to limit the present invention in any form. It should be noted that, for those skilled in the art, several variations and improvements can be made without departing from the scope of the present invention. These all fall within the scope of protection of the present invention.

[0030] 1) The "modified sodium alginate" described below in the present invention is prepared by the following method: 1 g of sodium alginate is dissolved in 100 mL of 2-morpholineethanesulfonic acid solution with a pH of 5.5 to a concentration of 0.1 wt %, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide and nitrogen-hydroxysuccinimide are added in a mass ratio of 2:1, and then 0.5 g of dopamine hydrochloride is added and reacted for 4 h to obtain modified sodium alginate.

[0031] 2) The "quaternized chitosan" described below in the present invention is a commercially available product. Example 1

[0032] This embodiment relates to a method for preparing a polyelectrolyte composite nanofiber membrane hemostatic auxiliary material prepared by a coaxial double-row electrospinning process, comprising the following steps:

[0033] 1. Dissolve polyvinyl alcohol in deionized water at 90°C to obtain a solution a with a concentration of 5 wt%;

[0034] 2. Dissolve the quaternized chitosan in an acetic acid aqueous solution to obtain a solution b with a concentration of 2 wt%;

[0035] 3. Dissolving the modified alginate in an aqueous solution to obtain a solution C with a concentration of 2 wt%;

[0036] 4. Electrospinning was performed using solution a obtained in step 1 as the core layer of the nanofiber, solution b obtained in step 2, and solution c obtained in step 3 as the shell layer of the nanofiber. The spinning temperature of the electrospinning was 37°C, the humidity was 20-30%, the distance between the syringe and the receiver (i.e., between the plates) was 20 cm, the voltage was 21 kV, the flow rate of the liquid in the double-row spinning was 0.09 µL / min, and the spinning time was 12 h to obtain polyelectrolyte composite nanofibers. Example 2

[0037] This embodiment relates to a method for preparing a polyelectrolyte composite nanofiber membrane hemostatic auxiliary material prepared by a coaxial double-row electrospinning process, comprising the following steps:

[0038] 1. Dissolve polyvinyl alcohol in deionized water at 90°C to obtain a solution a with a concentration of 7 wt%;

[0039] 2. Dissolve the quaternized chitosan in an acetic acid aqueous solution to obtain a solution b with a concentration of 2 wt %;

[0040] 3. Dissolving the modified alginate in an aqueous solution to obtain a solution C with a concentration of 2 wt%;

[0041] 4. Electrospinning was performed using solution a obtained in step 1 as the core layer of the nanofiber, solution b obtained in step 2, and solution c obtained in step 3 as the shell layer of the nanofiber. The spinning temperature of the electrospinning was 37°C, the humidity was 20-30%, the distance between the syringe and the receiver (i.e., between the plates) was 20 cm, the voltage was 21 kV, the flow rate of the liquid in the double-row spinning was 0.09 μL / min, and the spinning time was 12 h to obtain polyelectrolyte composite nanofibers. Example 3

[0042] This embodiment relates to a method for preparing a polyelectrolyte composite nanofiber membrane hemostatic auxiliary material prepared by a coaxial double-row electrospinning process, comprising the following steps:

[0043] 1. Dissolve polyvinyl alcohol in deionized water at 90°C to obtain a solution a with a concentration of 7 wt%;

[0044] 2. Dissolve the quaternized chitosan in an acetic acid aqueous solution to obtain a solution b with a concentration of 2 wt %;

[0045] 3. Dissolving the modified alginate in an aqueous solution to obtain a solution C with a concentration of 2 wt%;

[0046] 4. Solution a obtained in step 1 is used as the core layer of the nanofiber, solution b obtained in step 2, and solution c obtained in step 3 are used as the shell layer of the nanofiber, and electrospinning is performed. The spinning temperature of the electrospinning is 37°C, the humidity is 20-30%, the distance between the syringe and the receiver is 20 cm, the voltage is 21 kV, the flow rate of the liquid in the double-row spinning is 0.09 μL / min, and the spinning time is 12 h to obtain polyelectrolyte composite nanofibers. Example 4

[0047] This embodiment relates to a method for preparing a polyelectrolyte composite nanofiber membrane hemostatic auxiliary material prepared by a coaxial double-row electrospinning process, comprising the following steps:

[0048] 1. Dissolve polyvinyl alcohol in deionized water at 90°C to obtain a solution a with a concentration of 7 wt%;

[0049] 2. Dissolve the quaternized chitosan in an acetic acid aqueous solution to obtain a solution b with a concentration of 1 wt %;

[0050] 3. Dissolving the modified alginate in an aqueous solution to obtain a solution C with a concentration of 1 wt%;

[0051] 4. Solution a obtained in step 1 is used as the core layer of the nanofiber, solution b obtained in step 2, and solution c obtained in step 3 are used as the shell layer of the nanofiber, and electrospinning is performed. The spinning temperature of the electrospinning is 37°C, the humidity is 20-30%, the distance between the syringe and the receiver is 20 cm, the voltage is 21 kV, the flow rate of the liquid in the double-row spinning is 0.09 µL / min, and the spinning time is 12 h to obtain polyelectrolyte composite nanofibers. Comparative Example 1

[0052] At a temperature of 90°C, polyvinyl alcohol was dissolved in deionized water to obtain a solution with a concentration of 7 wt%, and electrospinning was performed. The electrospinning temperature was 37°C, the humidity was 20-30%, the voltage was 21 kV, the liquid flow rate was 0.09 µL / min, and the spinning time was 12 h to obtain a nanofabric containing only polyvinyl alcohol. Comparative Example 2

[0053] Polyvinyl alcohol was dissolved in deionized water at 90°C to obtain a solution with a concentration of 7 wt%, and modified alginate was dissolved at room temperature to obtain a solution with a concentration of 2 wt%. The two were then electrospun at a spinning temperature of 37°C, a humidity of 20-30%, a voltage of 21 kV, a liquid flow rate of 0.09 µL / min, and a spinning time of 12 h to obtain nanofabrics containing polyvinyl alcohol and modified alginate. Comparative Example 3

[0054] Polyvinyl alcohol was dissolved in deionized water at a temperature of 90°C to obtain a solution with a concentration of 7 wt%. Quaternized chitosan was dissolved in an acetic acid aqueous solution at room temperature to obtain a solution with a concentration of 2 wt%. The two were electrospun at a spinning temperature of 37°C, a humidity of 20-30%, a voltage of 21 kV, a liquid flow rate of 0.09 µL / min, and a spinning time of 12 h to obtain a nanofabric containing polyvinyl alcohol and quaternized chitosan. Example 5

[0055] This embodiment relates to a hemostatic effect test of a polyelectrolyte composite nanofiber membrane hemostatic auxiliary material prepared by a coaxial double-row electrospinning process, comprising the following steps:

[0056] 1. Anesthetize rats by intraperitoneal injection of 1 mL of 10 wt% chloral hydrate.

[0057] 2. Create a 1 cm × 0.05 cm injury model on the liver;

[0058] 3. Immediately cover the wound with polyelectrolyte composite nanofiber membrane hemostatic supplements (prepared in Example 4, Comparative Example 1, Comparative Example 2, and Comparative Example 3) of known area, medical gauze, and combat gauze to perform non-pressable model hemostasis for a short period of time;

[0059] 4. After 180 seconds, weigh the patient and calculate the amount of bleeding;

[0060] 5. The hemostatic effects of the control group gauze, combat gauze, polyvinyl alcohol nanofabric used in comparative example 1, quaternized chitosan nanofabric used in comparative example 2, modified alginate nanofabric used in comparative example 3 and polyelectrolyte composite nanofabric used in example 3 on rat liver were as follows: Figure 5 As shown. Compared with the control gauze, combat gauze, polyvinyl alcohol nanofabric used in comparative example 1, the polyvinyl alcohol and modified alginate nanofabric used in comparative example 2, and the nanofabric containing polyvinyl alcohol and quaternized chitosan used in comparative example 3, the hemostatic effect of the polyelectrolyte composite nanofabric used in Example 3 is greatly improved. This can be attributed to the electrostatic interaction between the two polyelectrolyte nanofibers when they encounter blood, thereby forming a dense network structure. At the same time, it has adhesion properties to tissues, can timely physically block the wound, enrich coagulation factors, and accelerate the hemostasis process, thus demonstrating its excellent hemostatic ability on non-pressable wounds.

[0061] The above describes the specific embodiments of the present invention. It should be understood that the present invention is not limited to the above specific embodiments, and those skilled in the art may make various variations or modifications within the scope of the claims, which do not affect the essence of the present invention.

Claims

1. A method for preparing a coaxial double-row polyelectrolyte composite nanofiber hemostatic auxiliary material, comprising the following steps: S1. dissolving polyvinyl alcohol in deionized water to obtain solution a; S2, dissolving the polyelectrolyte cation in an aqueous acetic acid solution to obtain solution b; S3, dissolving the polyelectrolyte anion in the aqueous solution to obtain solution c; S4, using the solution a obtained in step S1 as the core layer of the nanofiber, and the solution b obtained in step S2 and the solution c obtained in step S3 as the shell layer of the nanofiber, and performing electrospinning to obtain polyelectrolyte composite nanofibers; The polyelectrolyte anion is one of carboxymethyl chitosan, alginate, hyaluronic acid, and carboxymethyl cellulose, and the alginate is modified alginate; the polyelectrolyte cation is one of chitosan quaternary ammonium salt and chitosan, and the chitosan quaternary ammonium salt is quaternized chitosan; The added amounts of polyvinyl alcohol, quaternized chitosan, modified alginate, and acetic acid are 5-10%, 0.5-2.5%, 0.5-3%, and 50-90%, respectively; the added amounts are calculated as a percentage by mass of the total mixture of polyvinyl alcohol, quaternized chitosan, modified alginate, and acetic acid; The spinning temperature of electrospinning was 37°C, the humidity was 20-30%, the distance between the plates was 20 cm, the voltage was 21 kV, the flow rate of the liquid in the double-row spinning was 0.09 µL / min, and the spinning time was 12 h.

2. The method for preparing the coaxial double-row polyelectrolyte composite nanofiber hemostatic auxiliary material according to claim 1, characterized in that: The dissolution time of solution b and solution c is 8 h.

3. The method for preparing the coaxial double-row polyelectrolyte composite nanofiber hemostatic auxiliary material according to claim 1, characterized in that: The polyvinyl alcohol solution was dissolved in deionized water at 90°C, and the reaction time was 3 h.

4. A coaxial double-row polyelectrolyte composite nanofiber hemostatic adjuvant, which is obtained according to the preparation method according to any one of claims 1 to 3.

Images