Wound dressing carrying antibacterial components

Abstract

The invention provides a wound dressing carrying antibacterial components, namely a three-layer non-woven fabric drug-loaded dressing with asymmetric wettability, and the wound dressing has the advantages of rapid hemostasis, high antibacterial property and oxidation resistance, and can effectively promote wound healing. The circium japonicum extract and the antibacterial peptide loaded on the dressing provided by the invention enable the non-woven fabric to have high antibacterial, antioxidant and hemostatic properties, and the preparation method of the non-woven fabric is green and environment-friendly, high in safety and easy for large-scale production.

Description

Technical Field

[0001] The invention belongs to the technical field of biological products, and in particular relates to a wound dressing carrying antibacterial components. Background Art

[0002] Acute skin injuries often occur in life, but improper care can further deteriorate the wound, such as ulceration or scar formation. In addition to the ability to quickly stop bleeding, an ideal wound dressing must also provide continuous care for the wound. The hemostatic effect of medical cotton gauze is not significant, and when used for wound care, the gauze will also excessively absorb wound exudate, causing local dehydration of the wound and hindering the growth and repair of epithelial cells; excessive hydration and airtightness of hydrogel dressings and closed hydrophobic film dressings can easily lead to wound maceration. Moist healing can provide a moderately moist microenvironment for the wound, promote wound healing, and prevent scab formation. Therefore, the moisture management performance of wound dressings is particularly important.

[0003] By creating a difference in hydrophilicity in the liquid transmission direction of the fabric, the fabric can have asymmetric wetting properties. The needle punching process is a commonly used nonwoven fabric manufacturing and laminating process, and is widely used in the large-scale preparation of medical and sanitary fabrics. The hook of the needle punching machine drives the fibers to rub against each other and entangle, and can composite multiple layers of fabrics with different hydrophilicity. Summary of the invention

[0004] The purpose of the present invention is to provide a wound dressing carrying antibacterial components, namely a three-layer non-woven drug-carrying dressing with asymmetric wettability, which has rapid hemostasis, high antibacterial and antioxidant properties and can effectively promote wound healing.

[0005] The wound dressing provided by the present invention comprises the following steps:

[0006] 1) After the chitosan fiber is cut into 50mm short fibers, it is placed in a carding machine and a web laying machine for pretreatment. The treated short fibers are punched by a needle punch to form a surface density of 50±5g / m 2 Chitosan non-woven fabric.

[0007] 2) After the viscose fiber is cut into 80mm staple fibers, it is placed in a carding machine and a web laying machine for pretreatment. The treated staple fibers are punched through a needle punch to form a surface density of 80±5g / m 2 Viscose nonwoven fabric;

[0008] 3) After the polyester fiber is cut into 60mm staple fibers, it is placed in a carding machine and a web laying machine for pretreatment. The treated staple fibers are punched by a needle punching machine to form a surface density of 60±5g / m 2 Polyester non-woven fabric;

[0009] 4) stacking the chitosan non-woven fabric, the viscose non-woven fabric and the polyester non-woven fabric in sequence, and forming a three-layer non-woven fabric by a needle punching machine;

[0010] 5) Dissolving the Cirsium arvense extract in deionized water at a mass fraction of 2%-10% to prepare an aqueous solution of the Cirsium arvense extract, and spraying the aqueous solution of the Cirsium arvense extract onto the surface of the chitosan layer of the three-layer non-woven fabric after magnetic stirring to prepare the wound dressing of the present invention.

[0011] Preferably, the surface density of the inner chitosan nonwoven fabric in step 1) is 50±5 g / m 2 , 60±5g / m 2 , 70±5g / m 2 At least one of .

[0012] Preferably, in step 4), the surface density ratio of the chitosan layer, the viscose layer and the polyester layer in the three-layer non-woven fabric is 1-5:1-8:1-6;

[0013] Furthermore, antimicrobial peptides are also added in step 5);

[0014] A specific amino acid sequence of the antimicrobial peptide is LESWYVMNPEF (SEQ ID NO: 1).

[0015] The burdock extract and antimicrobial peptide loaded on the dressing provided by the present invention enable the non-woven fabric to have high antibacterial, antioxidant and hemostatic properties. The preparation method of the non-woven fabric is green and environmentally friendly, highly safe and easy to scale up. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 : Scanning electron microscope images of the present invention, (a) is a cross-sectional view of a chitosan-viscose-polyester three-layer non-woven fabric, (b) is the chitosan surface of a common three-layer non-woven fabric, and (c) is the drug-loaded chitosan surface.

[0017] Figure 2 : Hydrophobic performance diagram of ordinary three-layer non-woven fabric and drug-loaded three-layer non-woven fabric.

[0018] Figure 3 : Antioxidant performance diagram of ordinary three-layer non-woven fabric and drug-loaded three-layer non-woven fabric.

[0019] Figure 4 : Diagram of the antibacterial performance of screened peptides against Escherichia coli.

[0020] Figure 5 : Antibacterial performance diagram of drug-loaded non-woven fabric and drug-loaded and polypeptide-loaded non-woven fabric.

[0021] Figure 6 : Cytocompatibility diagram of drug-loaded nonwoven fabric and drug-loaded and polypeptide-loaded nonwoven fabric.

[0022] Figure 7 : Diagram of the hemostatic effect of drug-loaded non-woven fabric and drug-loaded and polypeptide-loaded non-woven fabric. DETAILED DESCRIPTION

[0023] The present invention is described in detail below in conjunction with specific embodiments and drawings.

[0024] Example 1: Preparation of drug-loaded three-layer non-woven fabric

[0025] 1. Preparation of single-layer non-woven fabric: After the chitosan fiber is cut into 60mm short fibers, it is placed in a carding machine and a web laying machine for pretreatment. The treated short fibers are punched by a needle punching machine to form a surface density of 50g / m 2 Chitosan nonwoven fabric with a surface density of 80g / m 2 Viscose nonwoven fabric with surface density 60g / m 2 Polyester non-woven fabric.

[0026] 2. The three single-layer non-woven fabrics are compounded by needle punching into a three-layer composite non-woven fabric.

[0027] 3. Prepare the water solution of Cirsium arvense extract: weigh 1g Cirsium arvense extract and dissolve it in 9g deionized water to prepare a 10% water solution of Cirsium arvense extract by mass.

[0028] 4. Loading of Cirsium arvense extract: Use SP202 ultrasonic sprayer to atomize Cirsium arvense extract solution and spray it on the chitosan layer of three-layer composite non-woven fabric, with a spraying speed of 0.5 ml / min, a spraying time of 50 min, and a spraying area of ​​10×10 cm 2 , carrier gas pressure 0.1 MPa, nozzle model FSW6002. The nonwoven fabric was dried in a drying oven at 40°C for 1 h.

[0029] Example 2: Determination of the properties of the drug-loaded three-layer non-woven fabric

[0030] 1. Morphological observation under electron microscope

[0031] Scanning electron microscopy was used to analyze the surface morphology of the common three-layer nonwoven fabric, the drug-loaded three-layer nonwoven fabric, and the cross-section of the drug-loaded three-layer nonwoven fabric. Figure 1 (a) is a cross-sectional view of a three-layer nonwoven fabric, (b) is a chitosan surface view of a three-layer nonwoven fabric, and (c) is a chitosan surface view of a three-layer nonwoven fabric loaded with Cirsium japonicum extract. The results show that the needle punching process successfully constructed a three-layer structure of the nonwoven fabric and successfully loaded Cirsium japonicum extract on chitosan fibers.

[0032] 2. Hydrophobicity

[0033] Take 3ml of blue ink and drop it on the three-layer composite non-woven fabric, and use the ultra-depth microscope to observe the surface and cross-section of the sample after the ink is dropped. Figure 2(a) is a picture of the chitosan layer after ink dropping, (b) is a picture of the polyester layer after ink dropping, and (a) is a cross-sectional picture of the three-layer non-woven fabric after ink dropping. The structure shows that only the middle viscose layer of the three-layer non-woven fabric absorbs moisture, and the chitosan layer in contact with the wound and the outermost polyester layer both maintain a good hydrophobic effect, which is beneficial to maintain moderate moisture on the wound surface and prevent external pollution from invading the wound.

[0034] 3. Antioxidant

[0035] The antioxidant activity of the prepared samples was evaluated by scavenging the stable chromogenic peroxidase substrate (ABTS) free radical. The results were analyzed by UV spectrophotometer. Figure 3 , Figure 3 This is the antioxidant efficiency diagram of ordinary three-layer non-woven fabric and drug-loaded three-layer non-woven fabric. The results show that the non-woven fabric loaded with thistle extract has excellent antioxidant effect.

[0036] Example 3: Preparation of polypeptides with antibacterial function

[0037] Take fresh and clean clam meat, mash it and grind it into paste, put the clam meat paste into a beaker and inject distilled water, add composite protease, use sodium hydroxide solution to adjust the solution pH to 8.0, and hydrolyze for 2 hours. After the reaction is completed, heat the enzymatic solution in a boiling water bath at 100°C for 10 minutes to inactivate it. After cooling, centrifuge the enzymatic solution at 8000r / min for 15 minutes, take the supernatant, dialyze it in a 500Da dialysis bag for 72 hours, and concentrate it under reduced pressure to make a dry powder.

[0038] The crude clam protease product was purified using a DEAE-Sepharose Fast Flow ion chromatography column. The crude polypeptide was dissolved in Tris-HCl buffer, centrifuged at 3000rmp for 5min, 10ml of the supernatant was added to the DEAE-Sepharose Fast Flow column, and then rinsed with Tris-HCl buffer, Tris-HCl buffer containing 0.1NaCl and 0.4mol solution. The peristaltic pump was adjusted to adjust the flow rate of the rinse liquid to 1.0m / min, and the rinse liquid was collected. A Sephadex G-25 gel chromatography column was used for secondary purification. The polypeptide purification component was dissolved in Tris-HCl buffer. After sufficient dissolution, it was centrifuged at 3000rmp for 5min, 2ml of the supernatant was taken, added to the Sephadex G-25 gel column, and rinsed with Tris-HCl buffer containing 0.1mol, NaCl solution. The peristaltic pump flow rate was adjusted to 1.0ml / min, and the rinse liquid was collected. The polypeptide sequence structure was then detected by UHPLC. The obtained polypeptide sequences were input into the BIOPEP-UWM bioactive peptide database for activity prediction, and the most active peptides were screened out, with the sequences of LESWYVMNPEF (polypeptide a), HGFEDDASPR (polypeptide b), HLNGAIKRPDS (polypeptide c) and HDSRPPDVKL (polypeptide d). They were sent to Shanghai Sangon Biotech Co., Ltd. for synthesis, and the antibacterial properties of the four polypeptides were tested using Escherichia coli. Figure 4 It can be seen that polypeptides c and d have no antibacterial activity against Escherichia coli, and polypeptide a has the strongest antibacterial activity. Therefore, polypeptide a is selected as the antimicrobial peptide to be added to the drug-loaded non-woven fabric, and its sequence is LESWYVMNPEF (SEQ ID NO: 1).

[0039] Example 4: Determination of the properties of drug-loaded and drug-loaded polypeptide-loaded three-layer non-woven fabrics

[0040] 1. Antibacterial

[0041] The drug-loaded nonwoven fabric and the drug-loaded and polypeptide-loaded nonwoven fabric were tested for Escherichia coli and Staphylococcus aureus using the dynamic contact method, and the antibacterial effect was analyzed by counting the number of colonies. Figure 5 The results showed that the non-woven fabric loaded with thistle extract had a certain antibacterial effect, and the antibacterial effect of the non-woven fabric was significantly enhanced after the addition of antimicrobial peptide (SEQ ID NO: 1).

[0042] 2. Biocompatibility

[0043] The biocompatibility of drug-loaded nonwoven fabrics and drug-loaded and polypeptide-loaded nonwoven fabrics was analyzed by MTT method. The results are shown in Figure 6 , the results showed that the extract of Cirsium japonicum and the antimicrobial peptide (SEQ ID NO: 1) did not affect the inherent excellent biocompatibility of the three-layer nonwoven fabric.

[0044] 3. Coagulation time

[0045] The hemostatic effects of drug-loaded nonwoven fabrics and drug-loaded polypeptide-loaded nonwoven fabrics were analyzed by coagulation time test. The results are shown in Figure 7 , with ordinary medical cotton yarn (Gauze) as the control, the results showed that the combined use of thistle extract and antimicrobial peptide (SEQ ID NO: 1) greatly improved the hemostatic effect of non-woven fabrics.

Claims

1. A wound dressing, It is characterized in that The preparation method of the wound dressing comprises the following steps: 1) After the chitosan fiber is cut into short fibers, the short fibers are sequentially placed in a carding machine and a web laying machine for pretreatment, and the treated short fibers are made into chitosan non-woven fabrics by a needle punching machine; 2) After the viscose fiber is cut into staple fibers, it is sequentially placed in a carding machine and a web laying machine for pretreatment, and the treated staple fibers are made into viscose nonwoven fabrics through a needle punching machine; 3) After the polyester fiber is cut into staple fibers, it is sequentially placed in a carding machine and a web laying machine for pretreatment; the treated staple fibers are made into polyester non-woven fabrics through a needle punching machine; 4) stacking the chitosan non-woven fabric, the viscose non-woven fabric and the polyester non-woven fabric in sequence, and forming a three-layer non-woven fabric by a needle punching machine; 5) dissolving the Cirsium arvense extract in deionized water to prepare an aqueous Cirsium arvense extract solution, and spraying the aqueous Cirsium arvense extract solution onto the surface of the chitosan layer of the three-layer non-woven fabric after magnetic stirring to prepare a wound dressing.

2. The wound dressing according to claim 1, It is characterized in that In the above 1), the chitosan fiber is cut into short fibers of 50 mm and made into a short fiber with a surface density of 50 ± 5 g / m 2 , 60±5g / m 2 or 70±5g / m 2 .

3. The wound dressing according to claim 1, It is characterized in that In the above 2), the viscose fiber is cut into 80 mm short fibers, and the surface density of the viscose non-woven fabric is 80 ± 5 g / m 2 .

4. The wound dressing according to claim 1, It is characterized in that 3) The polyester fiber is cut into 60 mm short fibers to obtain a surface density of 60 ± 5 g / m 2 Polyester non-woven fabric.

5. The wound dressing according to claim 1, It is characterized in that The surface density ratio of the chitosan layer, the viscose layer and the polyester layer in the three-layer non-woven fabric of step 4) is 1-5:1-8:1-6.

6. The wound dressing according to claim 1, It is characterized in that In the step 5), the cirsium japonicum extract is dissolved in deionized water at a mass fraction of 2-10%.

7. The wound dressing according to claim 1, It is characterized in that Antimicrobial peptides are also added to the aqueous solution of the burdock extract in step 5).

8. The wound dressing according to claim 1, It is characterized in that The amino acid sequence of the antimicrobial peptide is SEQ ID NO: 1.

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