Composite fiber membrane, preparation method and application thereof in diabetic wound dressing

Abstract

The invention belongs to the field of biomedical engineering, discloses a kind of composite fiber membrane, preparation method and application in its diabetic wound dressing.The preparation method prepares MOFs by hot solvent method and grows and is coated in nanofiber by spermidine, thioneine, tetra-(4-carboxylphenyl) porphine and copper nitrate, obtains composite fiber membrane.The composite fiber membrane is applied to diabetic wound dressing, can release spermidine and thioneine by blood response, accelerates and improves the formation of healthy blood vessels, promotes diabetic wound healing while controlling oxidative stress and anti-inflammatory.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical engineering, and in particular relates to a composite fiber membrane, a preparation method and application of the composite fiber membrane in diabetic wound dressings. Background Art

[0002] Diabetes can cause atherosclerosis and microvascular basement membrane thickening, glycogen deposition, and fatty and hyaline degeneration. Endothelial cells in blood vessels serve as a natural barrier between blood and the vessel wall. High blood sugar levels damage the endothelial structure, disrupting the integrity of the vessel wall. Damage to the endothelium exposes subendothelial collagen. To stop bleeding, platelets become activated, aggregating at the site of injury and forming a thrombus. These platelets attract fat deposits to the subendothelium, causing local narrowing of the blood vessels. Higher levels of glucose in the blood lead to increased blood concentration, slowing blood flow and causing stasis, which increases blood viscosity and promotes hypercoagulability. Furthermore, endothelial cell damage facilitates cholesterol deposition under the endothelium, forming atherosclerotic plaques and promoting the occurrence and progression of cardiovascular events. Increased lipid levels, thicker blood, and narrowed blood vessels all contribute to increased lipid deposition on the endothelium. If not properly controlled, this can clog the blood vessels, leading to the development of plaques and ultimately atherosclerosis. Blood vessels located in the area of ​​plaques experience decreased elasticity and increased brittleness, increasing the risk of rupture and potentially causing bleeding disorders.

[0003] Due to the existence of vascular disease mechanisms, the new blood vessels at the wound healing site of diabetic wounds are often poorly generated at the beginning and have basic defects, causing inflammatory factors to penetrate outside the blood vessels. Long-term chronic inflammation can seriously cause the wound to fail to heal. Summary of the Invention

[0004] In view of this, the object of the present invention is to provide a diabetic wound dressing and a preparation method thereof, which can release spermidine and ergothioneine through blood response, accelerate and improve the formation of healthy blood vessels, and promote diabetic wound healing while controlling oxidative stress and anti-inflammation.

[0005] In order to solve the above technical problems, the present invention provides the following technical solutions

[0006] A first aspect of the present invention provides a method for preparing a composite fiber membrane, comprising the following steps:

[0007] S1. Preparation of nanofiber membrane: Polyurethane and polyvinylpyrrolidone (PVP) were dissolved in a mixed solvent of N,N-dimethylformamide (DMF) and tetrahydrofuran (THF), and copper salt and meso-tetrakis(4-carboxyphenyl)porphine were added. The mixture was thoroughly stirred and ultrasonicated to achieve uniform dispersion. The mixture was shaken at 80-90°C for 12-20 hours to obtain a spinning solution, which was then electrospun to produce a nanofiber membrane.

[0008] S2. Treatment of the reaction solution: Immerse the nanofiber membrane obtained in step S1 in a 50-80 w / v% ethanol solution containing spermidine and thioneine, stir gently, and slowly add 10-15 w / v% sodium hydroxide solution dropwise to a solution pH of 9.5-10.0, and then continue for 24-48 hours. Take out the membrane, drain, and wash to obtain a composite fiber membrane.

[0009] Preferably, in step S1, in the mixed solvent of N,N-dimethylformamide and tetrahydrofuran, the volume ratio of N,N-dimethylformamide to tetrahydrofuran is 1:1.

[0010] Preferably, in step S1, the polyurethane is Carbothane aromatic thermoplastic polyurethane, model number is one of AC-4075A-B20, AC-4085A-B20 and AC-4095A-B20, and the concentration used is 9-12 w / v%.

[0011] Preferably, in step S1, the PVP is one of PVP-K25, PVP-K30 and PVP-K40, and the concentration used is 3-5 w / v%.

[0012] Preferably, in step S1, the copper salt is one of copper nitrate, copper chloride and copper sulfate, and the addition amount is 0.15-0.20 w / v%.

[0013] Preferably, in step S1, in the mixed solution, the concentration of polyurethane is 9-12 w / v%, the concentration of polyvinyl pyrrolidone is 3-5 w / v%, the concentration of copper salt is 0.15-0.20 w / v%, and the concentration of meso-tetrakis(4-carboxyphenyl)porphine is 0.62-0.83 w / v%.

[0014] Preferably, in step S1, the electrospinning conditions are voltage 16-20 kV, ambient humidity 20%-30% RH, ambient temperature 60-80°C, spinning distance 15-20 cm, and spinning solution flow rate 0.7-1.5 ml / h.

[0015] Preferably, in step S2, in the ethanol solution containing spermidine and thioneine, the concentration of spermidine is 0.14-0.28w / v%, and the concentration of thioneine is 1.80-2.40w / v%.

[0016] Preferably, in step S2, the bath ratio of the nanofiber membrane is 40-60:1.

[0017] Preferably, in step S2, adjusting the pH value to 9.5-10.0 is specifically performed by slowly dripping a sodium hydroxide solution until the pH value is 9.5-10.0; the concentration of the sodium hydroxide solution is 10-15 w / v%, and the dripping rate is 0.8-1.2 v / v% / min.

[0018] Preferably, in step S2, the washing specifically comprises: rinsing the composite fiber membrane with physiological saline for 3-5 times.

[0019] The second aspect of the present invention further provides a composite fiber membrane prepared by the above preparation method.

[0020] The third aspect of the present invention further provides the use of the composite fiber membrane in diabetic wound dressing.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] 1) The present invention constructs a polyurethane fiber with a structure of tetrakis(4-carboxyphenyl)porphine and Cu 2+ Based on the metal organic framework structure, Cu 2+ With multiple coordination numbers, spermidine and ergothioneine are then bound to the main body's Cu through the lone pair electrons on the amino, sulfhydryl, and carbon-sulfur groups. 2+ This can form a coordination adsorption. 2+ If the system contains Fe 2+ , Fe 2+ Can be used with Cu 2+ Competitive coordination binds spermidine and ergothioneine, allowing spermidine and ergothioneine to be released.

[0023] 2) Ergothioneine has antioxidant properties, preventing spermidine from being rapidly consumed by polyamine oxidase at the wound site, thereby increasing its bioavailability. Spermidine can promote protein synthesis or inhibit protein degradation. Spermidine also has neuroprotective effects; stimulates muscle protein synthesis, promoting muscle growth and repair; improves vascular elasticity, lowers blood pressure and cholesterol levels; and regulates the immune system, enhancing immunity and improving disease resistance. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a scanning electron microscope photograph of the diabetic wound dressing prepared in Example 1;

[0025] Figure 2 This is the curve of the healing rate changing with time in the diabetic wound healing experiment;

[0026] Figure 3 This is the relative expression result of VEGF in callus tissue on the 4th day of the diabetic wound healing experiment. DETAILED DESCRIPTION

[0027] In order to further understand the present invention, preferred embodiments of the present invention are described below in conjunction with examples. However, it should be understood that these descriptions are only for further illustrating the features and advantages of the present invention, rather than for limiting the claims of the present invention.

[0028] Example 1

[0029] A method for preparing a composite fiber membrane (diabetic wound dressing) comprises the following steps:

[0030] 1. Preparation of nanofiber membrane: Carbothane aromatic thermoplastic polyurethane AC-4085A-B20 and PVP-K30 were dissolved in a mixed solvent of equal volumes of DMF and THF at a polyurethane concentration of 10 w / v% and a PVP concentration of 4 w / v%. 0.18 w / v% copper nitrate and 0.78 w / v% of tetrakis(4-carboxyphenyl)porphine were then added. The mixture was thoroughly stirred and ultrasonicated to achieve uniform dispersion. The mixture was placed at 85°C for 18 h of oscillation reaction. The solution was then electrospun under the following conditions: voltage 18 kV, ambient humidity 25% RH, ambient temperature 70°C, spinning distance 17 cm, and spinning solution flow rate 1.0 ml / h to produce a nanofiber membrane.

[0031] 2. Reaction solution treatment: The nanofiber membrane prepared in step 1 was immersed in a 65w / v% ethanol solution containing 0.20w / v% spermidine and 2.00w / v% thioneine, with a bath ratio of 50:1, and gently stirred. A 12w / v% sodium hydroxide solution was added dropwise at a rate of 1v / v% / min until the solution pH value was 9.8. The reaction mixture was then continued for 36h, the membrane was taken out, and the composite fiber membrane was obtained by draining.

[0032] 3. Washing: Rinse the composite fiber membrane prepared in step 2 with normal saline for 4 times to obtain a diabetic wound dressing ( Figure 1 ).

[0033] Example 2

[0034] A method for preparing a composite fiber membrane (diabetic wound dressing) comprises the following steps:

[0035] 1. Preparation of nanofiber membrane: Carbothane aromatic thermoplastic polyurethane AC-4075A-B20 and PVP-K25 were dissolved in a mixed solvent of equal volumes of DMF and THF at a polyurethane concentration of 9 w / v% and a PVP concentration of 3 w / v%. 0.15 w / v% copper chloride and 0.62 w / v% of tetrakis(4-carboxyphenyl)porphine were then added. The mixture was thoroughly stirred and ultrasonicated to achieve uniform dispersion. The mixture was placed at 80°C for 20 h of oscillation reaction. The solution was then electrospun under the following conditions: a voltage of 16 kV, an ambient humidity of 20% RH, an ambient temperature of 60°C, a spinning distance of 15 cm, and a spinning solution flow rate of 0.7 ml / h to produce a nanofiber membrane.

[0036] 2. Reaction solution treatment: The nanofiber membrane prepared in step 1 was immersed in a 50w / v% ethanol solution containing 0.14w / v% spermidine and 1.80w / v% thioneine, with a bath ratio of 40:1, and gently stirred. A 10w / v% sodium hydroxide solution was added dropwise at a rate of 0.8v / v% / min until the solution pH value was 9.5. The reaction mixture was then continued for 24h, the membrane was taken out, and the composite fiber membrane was obtained by draining.

[0037] 3. Washing: Rinse the composite fiber membrane prepared in step 2 with physiological saline three times to obtain a diabetic wound dressing.

[0038] Example 3

[0039] A method for preparing a composite fiber membrane (diabetic wound dressing) comprises the following steps:

[0040] 1. Preparation of nanofiber membrane: Carbothane aromatic thermoplastic polyurethane AC-4095A-B20 and PVP-K40 were dissolved in a mixed solvent of equal volumes of DMF and THF at a polyurethane concentration of 12 w / v% and a PVP concentration of 5 w / v%. 0.20 w / v% copper sulfate and 0.83 w / v% of tetrakis(4-carboxyphenyl)porphine were then added. The mixture was thoroughly stirred and ultrasonicated to achieve uniform dispersion. The mixture was placed at 90°C for 12 h of oscillation reaction. The solution was then electrospun under the following conditions: a voltage of 20 kV, an ambient humidity of 30% RH, an ambient temperature of 80°C, a spinning distance of 20 cm, and a spinning solution flow rate of 1.5 ml / h to produce a nanofiber membrane.

[0041] 2. Reaction solution treatment: The nanofiber membrane prepared in step 1 is immersed in 80w / v% ethanol solution containing 0.28w / v% spermidine and 2.40w / v% thioneine, with a bath ratio of 60:1, and gently stirred. 15w / v% sodium hydroxide solution is added dropwise at a rate of 1.2v / v% / min until the solution pH value is 10.0, and then continued for 48h, the membrane is taken out, and drained to obtain a composite fiber membrane.

[0042] 3. Washing: Rinse the composite fiber membrane prepared in step 2 with physiological saline for 5 times to obtain a diabetic wound dressing.

[0043] Comparative Example 1 (without Cu-MOFs)

[0044] A method for preparing a dressing comprises the following steps:

[0045] 1. Nanofiber membrane preparation: Carbothane aromatic thermoplastic polyurethane AC-4085A-B20 and PVP-K30 were dissolved in a mixed solvent of equal volumes of DMF and THF at a polyurethane concentration of 10 w / v% and a PVP concentration of 4 w / v%. The mixture was thoroughly stirred and ultrasonically dispersed, and the mixture was oscillated at 85°C for 18 h. The solution was then electrospun under the following conditions: a voltage of 18 kV, an ambient humidity of 25% RH, an ambient temperature of 70°C, a spinning distance of 17 cm, and a spinning solution flow rate of 1.0 ml / h to produce a nanofiber membrane.

[0046] 2. Reaction solution treatment: The nanofiber membrane prepared in step 1 was immersed in a 65w / v% ethanol solution containing 0.20w / v% spermidine and 2.00w / v% thioneine, with a bath ratio of 50:1, and gently stirred. A 12w / v% sodium hydroxide solution was added dropwise at a rate of 1v / v% / min until the solution pH value was 9.8. The reaction mixture was then continued for 36h, the membrane was taken out, and the composite fiber membrane was obtained by draining.

[0047] 3. Washing: Rinse the composite fiber membrane prepared in step 2 with physiological saline for 4 times to obtain a dressing.

[0048] Comparative Example 2 (without thioneine)

[0049] A method for preparing a dressing comprises the following steps:

[0050] 1. Preparation of nanofiber membrane: Carbothane aromatic thermoplastic polyurethane AC-4085A-B20 and PVP-K30 were dissolved in a mixed solvent of equal volumes of DMF and THF at a polyurethane concentration of 10 w / v% and a PVP concentration of 4 w / v%. 0.18 w / v% copper nitrate and 0.78 w / v% of tetrakis(4-carboxyphenyl)porphine were then added. The mixture was thoroughly stirred and ultrasonicated to achieve uniform dispersion. The mixture was placed at 85°C for 18 h of oscillation reaction. The solution was then electrospun under the following conditions: voltage 18 kV, ambient humidity 25% RH, ambient temperature 70°C, spinning distance 17 cm, and spinning solution flow rate 1.0 ml / h to produce a nanofiber membrane.

[0051] 2. Treatment of reaction solution: Immerse the nanofiber membrane prepared in step 1 in a 65w / v% ethanol solution containing 0.20w / v% spermidine with a bath ratio of 50:1, stir gently, and add 12w / v% sodium hydroxide solution at a rate of 1v / v% / min until the solution pH is 9.8. Continue for 36 hours, remove the membrane, and drain to obtain a composite fiber membrane.

[0052] 3. Washing: Rinse the composite fiber membrane prepared in step 2 with physiological saline for 4 times to obtain a dressing.

[0053] Diabetes wound healing experiment

[0054] Eight-week-old male SD rats were selected and fed with a high-fat, high-sugar diet for 4 weeks. Type 2 diabetes was induced by a single intraperitoneal injection of 35 mg / kg of streptozotocin. The rats were randomly divided into six groups according to body weight using a random number table method, with five rats in each group. After anesthesia, hair was removed from both sides of the back. After disinfection with iodine, a circular wound with a diameter of approximately 1 cm was excised with a punch. The wounds were treated with the dressings prepared in Examples 1-3 and Comparative Examples 1-2, as well as commercially available dressings (WenJian Medical alginate dressing and Coloplast Kanghuier 3903 wound hydrogel dressing), respectively. The wounds were then wrapped and fixed with medical gauze. The wound area was recorded and the healing rate (wound area as a percentage of the original wound area) was calculated. The results are shown in Table 1. Figure 2 The same method was used to establish animal models. The animals were treated with the dressings prepared in Examples 1-3 and Comparative Examples 1-2, as well as commercially available dressings (WenJian Medical alginate dressing and Coloplast Kanghuier 3903 wound hydrogel dressing) for 4 days before being killed. The wound tissues were taken and the expression of VEGF was determined by Western blot (with β-actin as an internal reference). The statistical results are shown in Figure 3 .

[0055] Depend on Figure 2-Figure 3 It is known that the dressing prepared by the embodiment of the present invention has the effect of improving VEGF expression. VEGF, vascular endothelial growth factor, also known as vascular permeability factor, is a highly specific pro-vascular endothelial cell growth factor, which can reflect angiogenesis. The diabetic wound angiogenesis of the dressing processed by the embodiment is good, and abundant healthy blood vessels are conducive to the rapid healing of wounds. And in the comparative example 1 dressing, there is no Cu-MOFs formation, and the main structure with metal ions as the center is lacking in the fiber, and it is impossible to effectively coordinate and adsorb spermidine and thioneine, so the spermidine and thioneine contained in the dressing are less, more without blood responsiveness, and it is impossible to play the effect of promoting angiogenesis and wound healing. In the comparative example 2 dressing, there is no thioneine, when spermidine enters into tissue fluid, it can be metabolized by polyamine oxidase faster, and effective drug concentration cannot be maintained, then the formation of blood vessel and health status can be affected by high blood sugar, which is unfavorable for wound healing. In addition, two commercially available wound dressings are not taken measures for the new growth of healthy blood vessels, so wound healing effect is also not good.

[0056] The present invention provides a composite fiber membrane, a preparation method, and a method for its application in a diabetic wound dressing. Numerous methods and approaches exist for implementing this technical solution. The foregoing merely represents a preferred embodiment of the present invention. It should be noted that those skilled in the art may make various improvements and modifications without departing from the principles of the present invention, and such improvements and modifications are considered within the scope of the present invention. Components not specified in this embodiment may be implemented using existing technologies.

Claims

1. A method for preparing a composite fiber membrane, characterized in that: The steps include: S1. Preparation of nanofiber membrane: Polyurethane and polyvinylpyrrolidone were dissolved in a mixed solvent of N,N-dimethylformamide and tetrahydrofuran, and copper salt and meso-tetrakis(4-carboxyphenyl)porphine were added. The mixture was thoroughly stirred and ultrasonically dispersed to obtain a mixed solution. The mixed solution was oscillated at 80-90°C for 12-20 hours to obtain a spinning solution. The spinning solution was electrospun to produce a nanofiber membrane. S2. Treatment of the reaction solution: immersing the nanofiber membrane in an ethanol solution containing spermidine and thioneine, adjusting the pH value to 9.5-10.0, leaving it for 24-48 hours, then taking it out, draining it, and washing it to obtain a composite fiber membrane.

2. The preparation method according to claim 1, characterized in that In step S1, in the mixed solvent of N,N-dimethylformamide and tetrahydrofuran, the volume ratio of N,N-dimethylformamide to tetrahydrofuran is 1:

1.

3. The preparation method according to claim 1, characterized in that In step S1, the copper salt is one of copper nitrate, copper chloride and copper sulfate.

4. The preparation method according to claim 1, characterized in that In step S1, in the mixed solution, the concentration of polyurethane is 9-12 w / v%, the concentration of polyvinyl pyrrolidone is 3-5 w / v%, the concentration of copper salt is 0.15-0.20 w / v%, and the concentration of meso-tetrakis(4-carboxyphenyl)porphine is 0.62-0.83 w / v%.

5. The preparation method according to claim 1, characterized in that In step S1, the electrospinning conditions are as follows: voltage 16-20 kV, ambient humidity 20%-30% RH, ambient temperature 60-80° C., spinning distance 15-20 cm, and spinning solution flow rate 0.7-1.5 ml / h.

6. The preparation method according to claim 1, characterized in that In step S2, in the ethanol solution containing spermidine and thioneine, the concentration of spermidine is 0.14-0.28w / v%, and the concentration of thioneine is 1.80-2.40w / v%.

7. The preparation method according to claim 1, characterized in that In step S2, the bath ratio of the nanofiber membrane is 40-60:

1.

8. The preparation method according to claim 1, characterized in that In step S2, adjusting the pH value to 9.5-10.0 is specifically performed by slowly dripping a sodium hydroxide solution until the pH value is 9.5-10.0; the concentration of the sodium hydroxide solution is 10-15 w / v%, and the dripping rate is 0.8-1.2 v / v% / min.

9. A composite fiber membrane prepared according to the preparation method according to any one of claims 1 to 8.

10. Use of the composite fiber membrane according to claim 9 in diabetic wound dressing.

Images