CN116173289A - Functional dressing containing recombinant human collagen and preparation method and application thereof - Google Patents
Functional dressing containing recombinant human collagen and preparation method and application thereof Download PDFInfo
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- CN116173289A CN116173289A CN202211708397.0A CN202211708397A CN116173289A CN 116173289 A CN116173289 A CN 116173289A CN 202211708397 A CN202211708397 A CN 202211708397A CN 116173289 A CN116173289 A CN 116173289A
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- recombinant human
- human collagen
- dressing
- functional dressing
- stirring
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Links
- 102000008186 Collagen Human genes 0.000 title claims abstract description 87
- 108010035532 Collagen Proteins 0.000 title claims abstract description 87
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- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 48
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 48
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims abstract description 46
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- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims abstract description 19
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims abstract description 19
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims abstract description 19
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- DQJCDTNMLBYVAY-ZXXIYAEKSA-N (2S,5R,10R,13R)-16-{[(2R,3S,4R,5R)-3-{[(2S,3R,4R,5S,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}-5-(ethylamino)-6-hydroxy-2-(hydroxymethyl)oxan-4-yl]oxy}-5-(4-aminobutyl)-10-carbamoyl-2,13-dimethyl-4,7,12,15-tetraoxo-3,6,11,14-tetraazaheptadecan-1-oic acid Chemical compound NCCCC[C@H](C(=O)N[C@@H](C)C(O)=O)NC(=O)CC[C@H](C(N)=O)NC(=O)[C@@H](C)NC(=O)C(C)O[C@@H]1[C@@H](NCC)C(O)O[C@H](CO)[C@H]1O[C@H]1[C@H](NC(C)=O)[C@@H](O)[C@H](O)[C@@H](CO)O1 DQJCDTNMLBYVAY-ZXXIYAEKSA-N 0.000 description 6
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- 229940010747 sodium hyaluronate Drugs 0.000 description 1
- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical compound [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 description 1
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- 239000000126 substance Substances 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
- A61L26/0028—Polypeptides; Proteins; Degradation products thereof
- A61L26/0033—Collagen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
- A61L26/0023—Polysaccharides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
- A61L26/0028—Polypeptides; Proteins; Degradation products thereof
- A61L26/0047—Specific proteins or polypeptides not covered by groups A61L26/0033 - A61L26/0042
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0061—Use of materials characterised by their function or physical properties
- A61L26/0066—Medicaments; Biocides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/252—Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
- A61L2300/602—Type of release, e.g. controlled, sustained, slow
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Materials For Medical Uses (AREA)
Abstract
The invention belongs to the field of medical dressings, and particularly relates to a functional dressing containing recombinant human collagen, and a preparation method and application thereof. The functional dressing is a liquid dressing, and comprises the following raw materials in percentage by weight: 0.05-0.2% of recombinant human collagen, 1-5% of glycerol, 0.01-0.05% of edetate disodium, 0.01-0.05% of phenoxyethanol and purified water to 100%. The functional dressing of the invention may further comprise the following raw materials: 0.05-0.2% of thioglycopeptide, 0.5-1% of oxidized sodium alginate and 0.05-0.2% of trehalose. The functional dressing containing the recombinant human collagen has longer action time and obviously improves the action effects of resisting bacterial infection and promoting wound healing. In another aspect, the invention also provides the use of the functional dressing in the preparation of a medical dressing for repairing a skin wound.
Description
Technical Field
The invention belongs to the field of medical dressings, and particularly relates to a functional dressing containing recombinant human collagen, and a preparation method and application thereof.
Background
The skin is a barrier between the human body and the external environment, protecting various tissues and organs in the body from invasion by external physical, mechanical, chemical and pathogenic microorganisms. When the skin is damaged, if the wound cannot heal in time, the surface of the wound needs to be covered with the dressing in time. In addition, with the development of society, people pursue beauty more and more, so medical cosmetology is also becoming a popular consumption hot spot. At present, the researches on laser, fruit acid skin replacement, microcrystal grinding, water light and micro needle surgery are continuously and widely applied, and at the same time, after certain wound surface surgery is carried out, the skin epidermis is often damaged to different degrees, the skin barrier function is affected, and skin symptoms such as skin erythema, edema, scar, pigmentation and the like are easy to appear. Therefore, how to effectively repair the wound surface after laser, fruit acid skin replacement, microcrystalline grinding, water light and microneedle surgery becomes an important research hot spot.
The existing medical dressing medical appliance products for repairing the wound surface have single formula, and most of the products comprise single sodium hyaluronate or collagen, medical preservative and water for injection. Although the existing medical dressing can play a certain auxiliary repairing role on skin, can reduce the evaporation of moisture on the surface of the skin and play a role in moisturizing the skin, the existing medical dressing has insufficient effect on the aspects of resisting bacteria and infection, promoting wound healing and the like, lacks nursing function on skin wound surface, and has shorter normal acting time, so that the problems of long wound surface repairing period, easiness in secondary infection of the wound surface and the like are generally caused.
In view of the problems of poor antibacterial and anti-infection effects, promotion of wound healing and the like and short common action time of the medical dressing medical instruments for repairing the wound surface, which are used in the clinical and medical beauty fields at present, the inventor firstly provides an improved functional dressing containing recombinant human collagen for repairing the wound surface of the skin.
Disclosure of Invention
In order to overcome the defects in the prior art and solve the technical problems in the prior art, the invention provides a functional dressing containing recombinant human collagen, and a preparation method and application thereof. The functional dressing containing the recombinant human collagen has longer action time and obviously improves the action effects of resisting bacterial infection and promoting wound healing.
Specifically, the invention is realized through the following technical schemes:
in a first aspect, the present invention provides a functional dressing comprising recombinant human collagen, the functional dressing being a liquid dressing comprising the following raw materials in weight percent: 0.05 to 0.2 percent of recombinant human collagen, 1 to 5 percent of glycerol, 0.01 to 0.05 percent of edetate disodium, 0.01 to 0.05 percent of phenoxyethanol and purified water to 100 percent, and the functional dressing has the functions of resisting bacteria and infection and promoting wound healing.
Alternatively, in the above functional dressing, the recombinant human collagen is type III recombinant human collagen having a weight average molecular weight of greater than 10 kilodaltons.
Alternatively, in the above functional dressing, the recombinant human collagen has a weight average molecular weight of 11 to 14 kilodaltons.
Alternatively, in the above functional dressing, the functional dressing further comprises the following raw materials in percentage by weight: 0.05-0.2% of thioglycopeptide, 0.5-1% of oxidized sodium alginate and 0.05-0.2% of trehalose.
Preferably, in the above functional dressing, the functional dressing comprises the following raw materials: 0.15% recombinant human collagen, 3% glycerol, 0.04% disodium edentate, 0.01% phenoxyethanol, 0.2% glycopeptide, 1% oxidized sodium alginate and 0.15% trehalose, and purified water to 100%.
More preferably, in the above functional dressing, the functional dressing is made of the following raw materials: 0.15% recombinant human collagen, 3% glycerol, 0.04% disodium edentate, 0.01% phenoxyethanol, 0.2% glycopeptide, 1% oxidized sodium alginate and 0.15% trehalose, and purified water to 100%.
In an alternative way, in the functional dressing, the molecular weight of the oxidized sodium alginate is about 1.4 kilodaltons, and the oxidized sodium alginate presents a reticular porous structure and embeds the recombinant human collagen therein, so that the action time of the recombinant human collagen is prolonged, and a slow release effect is achieved.
Alternatively, in the above functional dressing, the preparation method of the oxidized sodium alginate includes the following steps: preparing 4g of sodium alginate into 4% aqueous solution by mass fraction, adding 1.5g of sodium periodate, oxidizing for 8 hours in a dark place, adding 2mL of glycol to terminate the oxidation reaction for 30 minutes, adding 2.5g of NaCl, fully mixing, adding 70mL of ethanol to precipitate and separate out, steaming, dissolving the product with deionized water, intercepting oxidized sodium alginate with molecular weight of more than 1.4 kilodaltons with a dialysis bag, changing deionized water every 6 hours, dialyzing for 24 hours, precipitating out with ethanol, filtering, and freeze-drying to obtain the sodium alginate.
In a second aspect, the present invention provides a method for preparing the functional dressing according to the first aspect, comprising the steps of:
step 1: injecting part of purified water into a first stirring container, then adding recombinant human collagen, if necessary, sequentially adding thioglycopeptide and trehalose, and stirring until all components are dissolved to obtain a first solution;
step 2: injecting the rest water into a second stirring container, adding oxidized sodium alginate if any, stirring until the oxidized sodium alginate is dissolved, sequentially adding glycerol, edetate disodium and phenoxyethanol, and stirring until all components are dissolved to obtain a second solution;
step 3: adding the first solution into the second solution while stirring, and continuously stirring until the mixture is uniformly mixed after the first solution is added to obtain a semi-finished functional dressing containing the recombinant human collagen;
step 4: and (3) placing the semi-finished functional dressing containing the recombinant human collagen for a period of time, filling the dressing into a sterile storage bottle through a filling machine, and finally performing electron beam sterilization and irradiation to obtain the functional dressing containing the recombinant human collagen.
Alternatively, in the above preparation method, the preparation method includes the steps of:
step 1: injecting part of purified water into a first stirring container, at the moment, the water temperature in the container is not higher than 35 ℃, then adding recombinant human collagen, if necessary, sequentially adding thioglycopeptide and trehalose, stirring at 20-80 rpm until each component is dissolved, and stirring for 10-30min to obtain a first solution;
step 2: injecting the rest water into a second stirring container, at the moment, if any, adding oxidized sodium alginate, stirring at 20-80 rpm for 10-30min until the water is not more than 35 ℃, then sequentially adding glycerol, disodium edentate and phenoxyethanol, stirring at 50-100 rpm until the components are dissolved, and stirring for 30-60min to obtain a second solution;
step 3: adding the first solution into the second solution under stirring, wherein the temperature in the container is not higher than 35 ℃, and continuously stirring at the rotation speed of 50rpm-100rpm until the mixture is uniformly mixed for 10-30min after the addition, so as to obtain a semi-finished functional dressing containing the recombinant human collagen;
step 4: placing the semi-finished functional dressing containing the recombinant human collagen at room temperature for 2 hours, filling the dressing into a sterile storage bottle through a filling machine, and finally performing electron beam sterilization irradiation for 1-10s to obtain the functional dressing containing the recombinant human collagen after sterilization.
In a third aspect, the invention provides the use of a functional dressing according to the first aspect or a functional dressing prepared by the preparation method according to the second aspect in the preparation of a medical dressing for repairing a skin wound.
Alternatively, in the above application, the medical dressing is used for laser, fruit acid skin replacement, microcrystalline grinding, water light needle or micro needle postoperative wound auxiliary repair, or for healing of acne damaged wound, for promoting repair of non-chronic wound and skin care.
Preferably, the application method of the medical dressing is to clean the skin of an affected part, directly spray or apply the medical dressing to the affected part, and uniformly apply the medical dressing, and the medical dressing is used every day or is in compliance with the doctor's advice.
It is understood that within the scope of the present invention, the above-described technical features of the present invention and technical features specifically described below (e.g., in the examples) may be combined with each other to constitute new or preferred technical solutions. Is limited to a space and will not be described in detail herein.
Compared with the prior art, the invention has the following beneficial effects:
(1) The invention provides a functional dressing containing recombinant human collagen, a preparation method and application thereof. The functional dressing containing the recombinant human collagen has longer action time and obviously improves the action effects of resisting bacterial infection and promoting wound healing.
(2) According to the invention, the viscosity of the dressing liquid can be prolonged by adding oxidized sodium alginate (with the molecular weight of about 14000 and no irritation), the residence time of the oxidized sodium alginate on the skin is prolonged, and the oxidized sodium alginate presents a reticular porous structure, so that the recombinant human collagen with a certain molecular weight range can be embedded in the oxidized sodium alginate, the action time of the recombinant human collagen is prolonged, and a slow release effect is achieved.
(3) The thioglycopeptide is added, is nontoxic and nonirritating, has the effects of treating mucosal injury, resisting bacteria and viruses and resisting inflammation, and has a synergistic effect with the recombinant collagen in repairing skin wound injury.
(4) The added trehalose has the effects of moisturizing and regulating viscosity, has better skin experience effect than the pure use of glycerol as a moisturizing agent for a subject, and reduces the sticky and greasy feeling of the liquid dressing when the liquid dressing is smeared on the skin.
Detailed Description
The invention will be further illustrated with reference to specific examples. It should be understood that the detailed description and specific examples are intended for purposes of illustration only and are not intended to limit the scope of the invention.
The specific techniques or conditions are not identified in the examples and are described in the literature in this field or are carried out in accordance with the product specifications. The reagents or equipment used were conventional products available for purchase through regular channels, with no manufacturer noted.
The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the examples described below, unless otherwise specified, are all commercially available products.
Preparation examples:
example 1
A functional dressing comprising recombinant human collagen, the functional dressing being a liquid dressing, the functional dressing being made from the following raw materials in weight percent: 0.15% recombinant human collagen, 3% glycerol, 0.04% disodium edentate, 0.01% phenoxyethanol, 0.2% glycopeptide, 1% oxidized sodium alginate and 0.15% trehalose, and purified water to 100%.
The recombinant human collagen is type III recombinant human collagen with weight average molecular weight of 11 ten thousand to 14 ten thousand daltons. The molecular weight of the oxidized sodium alginate is about 1.4 ten thousand daltons, and the preparation method of the oxidized sodium alginate comprises the following steps: preparing 4g of sodium alginate into 4% aqueous solution by mass fraction, adding 1.5g of sodium periodate, oxidizing for 8 hours in a dark place, adding 2mL of glycol to terminate the oxidation reaction for 30 minutes, adding 2.5g of NaCl, fully mixing, adding 70mL of ethanol to precipitate and separate out, steaming, dissolving the product with deionized water, intercepting oxidized sodium alginate with molecular weight of more than 1.4 kilodaltons with a dialysis bag, changing deionized water every 6 hours, dialyzing for 24 hours, precipitating out with ethanol, filtering, and freeze-drying to obtain the sodium alginate.
The preparation method of the functional dressing containing the recombinant human collagen comprises the following steps:
step 1: injecting part of purified water into a first stirring container, at the moment, the water temperature in the container is not higher than 35 ℃, then adding recombinant human collagen, if necessary, sequentially adding thioglycopeptide and trehalose, stirring at the rotating speed of 50rpm until all components are dissolved, and stirring for 20min to obtain a first solution;
step 2: injecting the rest water into a second stirring container, at the moment, if any, adding oxidized sodium alginate when the water temperature in the container is not higher than 35 ℃, stirring at the rotation speed of 50rpm until the sodium alginate is dissolved, stirring for 20min, sequentially adding glycerol, disodium edentate and phenoxyethanol, stirring at the rotation speed of 80rpm until each component is dissolved, and stirring for 40min to obtain a second solution;
step 3: adding the first solution into the second solution under stirring, wherein the temperature in the container is not more than 35 ℃, and continuously stirring at the rotation speed of 80rpm until the mixture is uniformly mixed for 20min after the addition, so as to obtain a semi-finished functional dressing containing the recombinant human collagen;
step 4: placing the semi-finished functional dressing containing the recombinant human collagen at room temperature for 2 hours, filling the dressing into a sterile storage bottle through a filling machine, and finally sterilizing the dressing by electron beam sterilization irradiation for 5 seconds to obtain the functional dressing containing the recombinant human collagen.
Comparative example 1
A functional dressing comprising recombinant human collagen, the functional dressing being a liquid dressing, the functional dressing being made from the following raw materials in weight percent: 0.35% recombinant human collagen, 3% glycerol, 0.04% disodium edentate, 0.01% phenoxyethanol, 1% oxidized sodium alginate, and 0.15% trehalose, and purified water to 100%.
The recombinant human collagen and oxidized sodium alginate used in comparative example 1 were prepared in the same manner as in example 1, and comparative example 1 was prepared in the same manner as in example 1.
Comparative example 2
A functional dressing comprising recombinant human collagen, the functional dressing being a liquid dressing, the functional dressing being made from the following raw materials in weight percent: 0.15% recombinant human collagen, 3% glycerol, 0.04% disodium edentate, 0.01% phenoxyethanol, 0.2% glycopeptide, 1% oxidized sodium alginate and 0.15% trehalose, and purified water to 100%.
The recombinant human collagen is type III recombinant human collagen with weight average molecular weight less than 5 ten thousand daltons. The oxidized sodium alginate used in comparative example 2 was the same as in example 1, and the preparation method of comparative example 2 was the same as in example 1.
Comparative example 3
A functional dressing comprising recombinant human collagen, the functional dressing being a liquid dressing, the functional dressing being made from the following raw materials in weight percent: 0.15% recombinant human collagen, 3% glycerol, 0.04% disodium edentate, 0.01% phenoxyethanol, 0.2% glycopeptide, 1% sodium alginate and 0.15% trehalose, and purified water to 100%.
The recombinant human collagen used in comparative example 3 was prepared in the same manner as in example 1, and the preparation method of comparative example 3 was the same as in example 1.
Comparative example 4
A functional dressing comprising recombinant human collagen, the functional dressing being a liquid dressing, the functional dressing being made from the following raw materials in weight percent: 0.3% recombinant human collagen, 3% glycerol, 0.04% disodium edentate, 0.01% phenoxyethanol, 0.2% glycopeptide and 1% oxidized sodium alginate, and purified water to 100%.
The recombinant human collagen and oxidized sodium alginate used in comparative example 4 were prepared in the same manner as in example 1, and comparative example 4 was prepared in the same manner as in example 1.
Effect examples:
the functional dressings prepared in example 1 and comparative examples 1-4 and containing recombinant human collagen were subjected to conventional animal skin irritation test, skin sensitization test and in vitro cytotoxicity test, respectively. The results show that the functional dressing containing recombinant human collagen is safe and nontoxic.
Effect example 1: physical properties of the functional dressing comprising recombinant human collagen according to the invention
Testing
pH value:
measured using a pH meter.
2. Viscosity:
the rotational viscometer measurement was carried out with a rotor number four at 3rpm.
The experimental results are shown in table 1 below.
Table 1: physical property test of functional dressing comprising recombinant human collagen according to the invention
| Group of | pH value of | Viscosity Pa.s |
| Example 1 | 6.48 | 139.27 |
| Comparative example 1 | 7.07 | 133.18 |
| Comparative example 2 | 6.42 | 121.52 |
| Comparative example 3 | 7.02 | 114.46 |
| Comparative example 4 | 6.55 | 119.28 |
The normal pH value of the skin surface is 4.5-6.5, and the wound healing is promoted under the weak acid condition, and from the results in table 1, it is known that example 1 and comparative examples 2 and 4 have weak acidity, and the pH of example 1 may more meet the requirement of the skin surface pH. In addition, the liquid dressing applied to the skin should have a suitable viscosity to ensure a relatively long duration of action, as can be seen from the results in table 1, the viscosity of example 1 is greater, which is advantageous for the functional dressing of the present invention. In addition, the experimental results of comparative examples 1-4 suggest that the molecular weight of recombinant human collagen, whether oxidized sodium alginate is used or not, and whether trehalose is used, have a large relationship with the viscosity of the functional dressing of the present invention.
Effect example 2: antibacterial experiments of functional dressing containing recombinant human collagen
After E.coli and Staphylococcus aureus are activated, strains are inoculated on an inclined plane, the inclined plane is cultured for 12 hours at 37 ℃, bacterial liquid on the inclined plane is washed by sterile water, OD value is measured to ensure that the bacterial amount of each inoculation is equivalent, bacterial liquid with equivalent bacterial amount is coated on a culture medium flat plate, filter paper sheets with the diameter of 0.6cm are placed in the flat plate by forceps, the liquid dressings of the example 1 and the comparative examples 1-4 are taken into small cylindrical gel with the same thickness by oxford cups and placed on the filter paper sheets, 3 parallel gradients are made for each gradient, the culture is carried out in a constant temperature incubator at 37 ℃ for 12 hours, and the average diameter of a bacteriostasis zone is measured.
Table 2: comparison of antimicrobial Properties of functional dressings comprising recombinant human collagen according to the invention
| Group of | Coli bacterium | Staphylococcus aureus |
| Example 1 | 1.3cm | 1.2cm |
| Comparative example 1 | 0.6cm | 0.6cm |
| Comparative example 2 | 0.9cm | 1.0cm |
| Comparative example 3 | 1.2cm | 1.1cm |
| Comparative example 4 | 1.2cm | 1.2cm |
The results in table 2 show that example 1 shows a stronger effect against escherichia coli and staphylococcus aureus than comparative examples 1 and 2, which suggests that both the thioglycopeptides contained in the functional dressing comprising recombinant human collagen of the present invention and the recombinant human collagen of a specific molecular weight range may have an affinity with the antibacterial effect of the functional dressing, in particular that the contribution of the thioglycopeptides to the antibacterial effect is greater. From the results of comparative examples 3 and 4, it can be seen that whether oxidized sodium alginate is used and whether trehalose is added has little effect on the antimicrobial properties of the functional dressing of the present invention.
Effect example 3: the invention relates to a wound repair experiment of a functional dressing containing recombinant human collagen
1. The experimental object:
randomly selecting 120 patients with the face just after the microneedle operation, 30 men and 90 women with the age of 18-55 years, randomly dividing the patients into 6 groups, wherein the basic data of each patient has no obvious difference, and p is more than 0.05, and comparing the basic data of each patient;
2. the treatment method comprises the following steps:
after daily skin lesion site cleansing, the skin wound dressing of each experimental group (example 1, comparative examples 1 to 4, and positive control commercially available commercial product "human recombinant collagen gel dressing") was applied 1 time at night during days 1 to 10, 1 time at night during days 10 to 20, and 20 minutes each time at intervals;
3. treatment criteria:
3.1 skin symptom score: erythema, edema, exudation, stinging, pigmentation were 0-3 minutes in terms of none, light, medium and heavy, respectively; the specific criteria are as follows:
erythema: red spot color is 3 minutes; obvious erythema is counted for 2 minutes; 1 minute of red spot lightening meter; no color change was counted for 0 point.
Edema: the extensive area is subjected to dermal edema infiltration gauge for 3 minutes; well-defined dermal oedema occurred at multiple points, 2 points; local area dermal edema is not easily divided by tactometer 1; no edema was counted as 0 point.
Oozing: exudation was evident, 3 minutes in the scale; exudation is more obvious, and the range is smaller by 2 minutes; after careful observation, a punctiform exudation meter was seen for 1 minute; no exudation was measured for 0 min.
Stinging: the pain is continuously felt, and 3 minutes can not be counted; continuous or intermittent tingling sensation can be counted for 2 minutes; occasionally, a slight stinging score of 1; the pain sensation without thorns is counted for 0 minutes.
Pigmentation: pigmentation was evident, with a range of 3 minutes; pigmentation was more pronounced, the range was less 2 minutes; a slight pigmentation was seen 1 minute after careful observation; no pigmentation was counted at 0 point.
Efficacy evaluation criteria: the traditional Chinese medicine is divided into four criteria of cure, obvious effect, effective and ineffective.
Efficacy index = [ (pre-treatment integral-post-treatment integral)/(pre-treatment integral ] ×100%).
And (3) healing: the curative effect index is more than or equal to 95%; the effect is shown: the curative effect index is 70% -94%; the method is effective: 30% -69% of curative effect index; invalidation: the curative effect index is less than 30 percent. Total effective rate = (number of recovery cases + number of significant cases + number of effective cases)/(total cases x 100%).
3.2 time for wound healing: the average wound healing time (days) of patients in each experimental group and control group was counted separately
3.3 skin physiological function index detection: the non-invasive skin physiological function tester is adopted to detect the water content of the stratum corneum, the percutaneous water loss (TEWL) and the skin grease content of patients in each experimental group, and the testing method comprises the following steps: and (3) measuring under the conditions of constant temperature and constant humidity, wherein the room temperature is 23-26 ℃, the relative humidity is 40% -60%, respectively detecting non-skin damage parts of cheeks of a tested person, cleaning skin to be tested before detection, and entering a test environment for 30min in advance for testing.
The statistical test method comprises the following steps: the results were analyzed using SPSS 23.0 statistical software and the data are expressed as mean.+ -. Standard deviation. The comparison between groups uses a one-factor analysis of variance, and p <0.05 indicates that the difference is statistically significant.
4. Treatment results:
4.1 skin symptoms and average wound healing time results are shown in table 3 below:
table 3: skin symptoms and average wound healing time
The results in table 3 show that the subjects using example 1 showed a significant improvement in skin symptom score and average wound healing time compared to comparative examples 1-4 and positive control, with a total effective rate of 100% and an average wound healing time shortened to 3 days.
From the experimental results of comparative examples 1 to 4, it can be seen that whether the functional dressing comprising recombinant human collagen of the present invention contains the thioglycopeptide has a very important effect on skin symptoms and average wound healing time, and when the functional dressing does not contain the thioglycopeptide, the total effective rate is only 40%, and the average wound healing time is as long as 11 days. In addition, the recombinant human collagen with a specific molecular weight range and whether oxidized sodium alginate is used have a relatively obvious effect on the functional dressing comprising the recombinant human collagen, but the effect of trehalose on the functional dressing comprising the recombinant human collagen is relatively less.
4.2 at the end of treatment, the results of percutaneous water loss (TEWL), stratum corneum moisture content and skin lipid content are shown in table 4 below:
table 4: percutaneous Water loss (TEWL), stratum corneum moisture content and skin oil content
Note that: * Comparison with positive control group p <0.05, comparison with positive control group p <0.01.
The results in table 4 show that subjects using example 1 showed significant improvement in skin barrier function compared to comparative examples 1-4 and positive controls after 20 days of treatment with the skin wound repair liquid dressing of the present invention. The results of the example 1 subjects were significantly better than the positive control (p < 0.01) for both the percutaneous water loss, the stratum corneum water content and the skin oil content.
From the experimental results of comparative examples 1 to 4, it can be seen that whether the functional dressing of the present invention comprising recombinant human collagen contains a thioglycopeptide has a very important effect on skin water loss, stratum corneum water content and skin oil content, and the above index is significantly deteriorated when the thioglycopeptide is not contained (see the result of comparative example 1 for details). In addition, the recombinant human collagen with a specific molecular weight range and whether oxidized sodium alginate is used or not have a more remarkable influence on the indexes of skin water loss, stratum corneum water content and skin fat content of the functional dressing containing the recombinant human collagen, but the influence of trehalose on the effect of the functional dressing containing the recombinant human collagen is relatively small.
It will be apparent to those skilled in the art that various modifications and variations can be made to the present invention without departing from the spirit or scope of the invention. Thus, it is intended that the present invention also include such modifications and alterations insofar as they come within the scope of the appended claims or the equivalents thereof.
Claims (10)
1. A functional dressing comprising recombinant human collagen, characterized in that: the functional dressing is a liquid dressing, and comprises the following raw materials in percentage by weight: 0.05 to 0.2 percent of recombinant human collagen, 1 to 5 percent of glycerol, 0.01 to 0.05 percent of edetate disodium, 0.01 to 0.05 percent of phenoxyethanol and purified water to 100 percent, and the functional dressing has the functions of resisting bacteria and infection and promoting wound healing.
2. The functional dressing of claim 1, wherein: the recombinant human collagen is type III recombinant human collagen with weight average molecular weight of more than 10 ten thousand daltons.
3. The functional dressing of claim 2, wherein: the recombinant human collagen has a weight average molecular weight of 11 to 14 kilodaltons.
4. A functional dressing according to any one of claims 1 to 3, wherein: the functional dressing further comprises the following raw materials in percentage by weight: 0.05-0.2% of thioglycopeptide, 0.5-1% of oxidized sodium alginate and 0.05-0.2% of trehalose.
5. The functional dressing of claim 4, wherein: the molecular weight of the oxidized sodium alginate is about 1.4 ten thousand daltons, the oxidized sodium alginate presents a reticular porous structure and embeds the recombinant human collagen therein, so that the action time of the recombinant human collagen is prolonged, and the slow release effect is achieved.
6. The functional dressing of claim 4 or claim 5, wherein: the preparation method of the oxidized sodium alginate comprises the following steps: preparing 4g of sodium alginate into 4% aqueous solution by mass fraction, adding 1.5g of sodium periodate, oxidizing for 8 hours in a dark place, adding 2mL of glycol to terminate the oxidation reaction for 30 minutes, adding 2.5g of NaCl, fully mixing, adding 70mL of ethanol to precipitate and separate out, steaming, dissolving the product with deionized water, intercepting oxidized sodium alginate with molecular weight of more than 1.4 kilodaltons with a dialysis bag, changing deionized water every 6 hours, dialyzing for 24 hours, precipitating out with ethanol, filtering, and freeze-drying to obtain the sodium alginate.
7. A method of preparing a functional dressing according to any one of claims 1 to 6, characterized in that: the method comprises the following steps:
step 1: injecting part of purified water into a first stirring container, then adding recombinant human collagen, if necessary, sequentially adding thioglycopeptide and trehalose, and stirring until all components are dissolved to obtain a first solution;
step 2: injecting the rest water into a second stirring container, adding oxidized sodium alginate if any, stirring until the oxidized sodium alginate is dissolved, sequentially adding glycerol, edetate disodium and phenoxyethanol, and stirring until all components are dissolved to obtain a second solution;
step 3: adding the first solution into the second solution while stirring, and continuously stirring until the mixture is uniformly mixed after the first solution is added to obtain a semi-finished functional dressing containing the recombinant human collagen;
step 4: and (3) placing the semi-finished functional dressing containing the recombinant human collagen for a period of time, filling the dressing into a sterile storage bottle through a filling machine, and finally performing electron beam sterilization and irradiation to obtain the functional dressing containing the recombinant human collagen.
8. The method of manufacturing according to claim 7, wherein: the method comprises the following steps:
step 1: injecting part of purified water into a first stirring container, at the moment, the water temperature in the container is not higher than 35 ℃, then adding recombinant human collagen, if necessary, sequentially adding thioglycopeptide and trehalose, stirring at 20-80 rpm until each component is dissolved, and stirring for 10-30min to obtain a first solution;
step 2: injecting the rest water into a second stirring container, at the moment, if any, adding oxidized sodium alginate, stirring at 20-80 rpm for 10-30min until the water is not more than 35 ℃, then sequentially adding glycerol, disodium edentate and phenoxyethanol, stirring at 50-100 rpm until the components are dissolved, and stirring for 30-60min to obtain a second solution;
step 3: adding the first solution into the second solution under stirring, wherein the temperature in the container is not higher than 35 ℃, and continuously stirring at the rotation speed of 50rpm-100rpm until the mixture is uniformly mixed for 10-30min after the addition, so as to obtain a semi-finished functional dressing containing the recombinant human collagen;
step 4: placing the semi-finished functional dressing containing the recombinant human collagen at room temperature for 2 hours, filling the dressing into a sterile storage bottle through a filling machine, and finally performing electron beam sterilization irradiation for 1-10s to obtain the functional dressing containing the recombinant human collagen after sterilization.
9. Use of the functional dressing according to any one of claims 1 to 6 or the functional dressing prepared by the preparation method according to claim 7 or claim 8 for preparing a medical dressing for repairing skin wound.
10. Use according to claim 9, characterized in that: the medical dressing is used for auxiliary repair of wound surfaces after laser, fruit acid skin replacement, microcrystalline grinding, water light needle or microneedle operation or healing of damaged wound surfaces of acne, and is used for promoting repair of non-chronic wound surfaces and nursing of skin.
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