CN117100905A - Dressing for promoting wound healing - Google Patents

Dressing for promoting wound healing Download PDF

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Publication number
CN117100905A
CN117100905A CN202311384551.8A CN202311384551A CN117100905A CN 117100905 A CN117100905 A CN 117100905A CN 202311384551 A CN202311384551 A CN 202311384551A CN 117100905 A CN117100905 A CN 117100905A
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CN
China
Prior art keywords
dressing
growth factor
oxygen
cell growth
hemoglobin
Prior art date
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Pending
Application number
CN202311384551.8A
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Chinese (zh)
Inventor
李文亮
张彦鹏
白珂
陈浩源
张天化
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Runfang Beijing Biotechnology Co ltd
Redpharm Beijing Biomedical Research Institute Co ltd
Original Assignee
Runfang Beijing Biotechnology Co ltd
Redpharm Beijing Biomedical Research Institute Co ltd
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Publication date
Application filed by Runfang Beijing Biotechnology Co ltd, Redpharm Beijing Biomedical Research Institute Co ltd filed Critical Runfang Beijing Biotechnology Co ltd
Priority to CN202311384551.8A priority Critical patent/CN117100905A/en
Publication of CN117100905A publication Critical patent/CN117100905A/en
Pending legal-status Critical Current

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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L26/00Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0009Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
    • A61L26/0028Polypeptides; Proteins; Degradation products thereof
    • A61L26/0047Specific proteins or polypeptides not covered by groups A61L26/0033 - A61L26/0042
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L26/00Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0061Use of materials characterised by their function or physical properties
    • A61L26/0066Medicaments; Biocides
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/795Porphyrin- or corrin-ring-containing peptides
    • C07K14/805Haemoglobins; Myoglobins
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L2300/00Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/20Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
    • A61L2300/252Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L2300/00Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/404Biocides, antimicrobial agents, antiseptic agents
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L2300/00Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/412Tissue-regenerating or healing or proliferative agents
    • A61L2300/414Growth factors

Abstract

The invention discloses a dressing for promoting wound healing, which comprises a cell growth factor and an oxygen carrier protein. In the dressing, the cell growth factors are combined with the receptors on the cell surfaces to activate signal paths in the cells, promote DNA synthesis and cell division and proliferation of the cells, and promote the growth and proliferation of the cells, while the oxygen-carrying proteins can provide effective oxygen supply for the growth and proliferation of the cells, and under the synergistic cooperation of the cell growth factors and the oxygen-carrying proteins, the healing of the wound surfaces difficult to heal can be effectively promoted. The cell growth factor is taken as a protein factor, the use of the cell growth factor is limited by the high price, and the addition amount of the cell growth factor can be greatly reduced through the cooperative coordination of the oxygen-carrying proteins, so that the aim of synergistically accelerating the healing of chronic wound surfaces is fulfilled while the minimum addition amount of the cell growth factor with effectiveness is greatly reduced. In combination with the test example part, the dressing provided by the invention can effectively promote the accelerated healing of diabetic foot ulcers (difficult-to-heal wound surfaces).

Description

Dressing for promoting wound healing
Technical Field
The invention relates to the technical field of dressing of medical equipment, in particular to a dressing for promoting wound healing.
Background
The wound surface difficult to heal is not uniformly defined at present, and the shape, size and depth of the wound surface cannot be uniformly defined due to various causes such as trauma, infection, tumor or nodule burst. It is generally considered that a difficult-to-heal wound is a chronic wound that is not healed or prone to healing after 4 weeks of standard treatment due to various etiologies. At present, the population of society tends to be aged, the base number of the population of hyperglycemia, hypertension and hyperlipidemia is larger and larger, and the number of related difficult-to-heal wound patients is increased year by year. Although the difficult-to-heal wound surface does not immediately threaten the life safety of the patient, the physical and mental health and the quality of life of the patient are greatly influenced, and the household economic burden is increased. Although a plurality of methods are clinically used for treating the difficult-to-heal wound surface, the traditional treatment method has very little effect on the complicated difficult-to-heal wound surface, and the search for a new effective method for promoting the healing of the difficult-to-heal wound surface is an important subject for common research and exploration in the multidisciplinary field.
The serious injury of local cells of the wound surface causes that the number of repair seed cells remained on the local surface of the wound is small, the function is dysfunctional or low, the local blood supply of the wound surface is poor, and sufficient nutrition is difficult to provide to meet the needs of regeneration and repair of local tissue cells, and along with the rapid development of gene therapy and stem cell engineering technology, the practice proves that the cell growth factors can be separated and are applied to an important tissue repair cell of which the reaction of the microenvironment of the wound surface participates in the healing of skin wound. The p value of the cell growth factor is 5.8, is consistent with the pH value of the complete skin of a human body, can effectively protect the slightly acidic environment of the skin and prevent bacteria from generating, but the pH value of the skin with wound surface can reach about 7.4, the skin can be yellow after long-term use, and the repair depth is limited. The wound surface difficult to heal is generally infected and has abscess, and in this aspect, the cell growth factors cannot function.
Oxygen therapy of wound surfaces has been studied for many years, and oxygen has the effects of sterilizing the wound surfaces, promoting cell repair, avoiding infection and the like. 2003. In the years, sen et al first proposed that the main cause of delayed wound healing is the occurrence of hypoxia in the wound and treatment measures such as wound oxygen therapy, and ensuring normal oxygenation of the wound is considered to be a key node for promoting wound closure. From the physiological condition, a large amount of oxygen is needed in the process of closing the wound surface, and the wound surface can cause a low-oxygen environment, so that the low-oxygen stimulates tissue regeneration. Clinically, hyperbaric oxygen is often adopted to treat the wound surface difficult to heal, and the oxygen content of the whole body tissue is increased to improve the local anoxic state and promote the wound surface to heal. The method is controversial, patients can cause oxygen poisoning, barotrauma, decompression sickness and other side effects, and hyperbaric oxygen also has a plurality of contraindications, so that patients with wound surfaces difficult to heal cannot benefit from oxygen therapy.
Disclosure of Invention
Based on this, there is a need to provide a dressing that promotes wound healing that can solve the above-mentioned problems.
A dressing for promoting wound healing comprising a cell growth factor and an oxygen carrier.
In one embodiment, the mass ratio of the cell growth factor to the oxygen carrier protein is 1-20: 30-130.
In one embodiment, the dressing is a liquid dressing;
the concentration of the cell growth factor is 10 mg/L-200 mg/L, and the concentration of the oxygen carrier protein is 300 mg/L-1300 mg/L.
In one embodiment, the cell growth factor is selected from at least one of an epidermal growth factor, a fibroblast growth factor, a keratinocyte growth factor, a vascular endothelial growth factor, a transforming growth factor, and a platelet-derived growth factor.
In one embodiment, the oxygen carrying protein is selected from at least one of hemoglobin, modified hemoglobin, myoglobin, and modified myoglobin.
In one embodiment, the modified hemoglobin is prepared by:
extracting red blood cells from whole blood of an animal;
fully mixing the red blood cells with a diluent to completely lyse the red blood cells, thereby obtaining a hemoglobin lysate;
percolating, purifying and degerming the hemoglobin lysate to obtain a hemoglobin solution;
mixing the hemoglobin solution with a protein modifier for reaction to obtain the modified hemoglobin;
alternatively, the modified myoglobin is prepared by the following operations:
extracting skeletal muscle from skeletal muscle to obtain skeletal muscle extract;
percolating, purifying and sterilizing the skeletal muscle extract to obtain myoglobin solution;
and mixing and reacting the myoglobin solution with a protein modifier to obtain the modified myoglobin.
In one embodiment, the protein modifier is selected from at least one of glutaraldehyde, glyoxal, bis-aspirin, polyethylene glycol, 1, 5-hexadiene, dextran, starch slurry, or serum albumin.
In one embodiment, the dressing further comprises an adjunct;
the auxiliary materials are selected from at least one of electrolyte, adhesive, antioxidant, analgesic, osmotic pressure regulator, stabilizer, bacteriostat, pH value regulator, flocculating agent and film forming agent.
In one embodiment, the content of the oxo-oxygen carrier protein is less than or equal to 60 percent.
In one embodiment, the content of the oxo-oxygen carrier protein is less than or equal to 10 percent.
The dressing for promoting wound healing comprises cell growth factors and oxygen-carrying proteins, wherein the cell growth factors are combined with receptors on the surfaces of cells to activate signal paths in the cells, promote DNA synthesis and cell division and proliferation of the cells, and can promote the growth and proliferation of the cells, and the oxygen-carrying proteins can provide effective oxygen supply for the growth and proliferation of the cells, so that the healing of difficult-to-heal wound surfaces can be effectively promoted under the synergistic cooperation of the cell growth factors and the oxygen-carrying proteins. The cell growth factor is taken as a protein factor, the use of the cell growth factor is limited by the high price, and the addition amount of the cell growth factor can be greatly reduced through the cooperative coordination of the oxygen-carrying proteins, so that the aim of synergistically accelerating the healing of chronic wound surfaces is fulfilled while the minimum addition amount of the cell growth factor with effectiveness is greatly reduced.
In combination with the test example part, it can be seen that the dressing for promoting wound healing provided by the invention can effectively promote the accelerated healing of diabetic foot ulcers (difficult-to-heal wounds).
In addition, the wound surface of the difficult-to-heal wound surface has complex cause, blood is often entrained and inflammatory reaction occurs, the inflammatory reaction also needs energy supply, and oxygen released by the oxygen carrier protein added into the dressing for promoting wound healing can participate in the inflammatory reaction, and the oxygen carrier protein and the cell growth factor can promote the wound surface to heal in an accelerating way.
In addition, in the wound healing process, the growth environment of the skin can be changed, so that bacterial infection, especially anaerobic bacteria are generated in a large amount, and the oxygen-carrying protein added into the dressing for promoting wound healing, unlike hyperbaric oxygen, has good inhibition and killing effects on anaerobic bacteria infection, and can promote wound healing.
Preferably, compared with erythrocytes carrying oxygen in the same way, the oxygen carrying protein is a nanoscale protein, has the advantages of no need of matching, wide working temperature range (2-40 ℃), no coagulation risk, stability of 36 months and the like, and has the oxygen carrying capacity 7-9 times that of erythrocytes.
Preferably, the dressing for promoting wound healing is a liquid dressing, and small-dose local perfusion can not bring perfusion damage to the wound surface, but can relieve ischemia caused by blood vessels of the wound surface or ischemia caused by vascular occlusion caused by inflammation.
The dressing for promoting wound healing can be packaged in spray bottles for use, has the characteristics of no irritation, extremely low toxicity, quick response and long-term storage, can promote the repair and regeneration of tissue cells, reduce the probability of infection, accelerate the healing speed of wound surfaces, and simultaneously relieve the feeling of itching and discomfort.
The dressing for promoting wound healing provided by the invention has the advantages of small volume, easiness in carrying and convenience in use, is suitable for various scenes, can be used for self-treatment outside a hospital, and is easily sprayed on a wound to promote wound healing.
Detailed Description
The following description of the embodiments of the present invention will clearly and fully describe the technical solutions of the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The invention discloses a dressing for promoting wound healing, which comprises a cell growth factor and an oxygen carrier protein.
The dressing for promoting wound healing comprises cell growth factors and oxygen-carrying proteins, wherein the cell growth factors are combined with receptors on the surfaces of cells to activate signal paths in the cells, promote DNA synthesis and cell division and proliferation of the cells, and can promote the growth and proliferation of the cells, and the oxygen-carrying proteins can provide effective oxygen supply for the growth and proliferation of the cells, so that the healing of difficult-to-heal wound surfaces can be effectively promoted under the synergistic cooperation of the cell growth factors and the oxygen-carrying proteins. The cell growth factor is taken as a protein factor, the use of the cell growth factor is limited by the high price, and the addition amount of the cell growth factor can be greatly reduced through the cooperative coordination of the oxygen-carrying proteins, so that the aim of synergistically accelerating the healing of chronic wound surfaces is fulfilled while the minimum addition amount of the cell growth factor with effectiveness is greatly reduced.
In combination with the test example part, it can be seen that the dressing for promoting wound healing provided by the invention can effectively promote the accelerated healing of diabetic foot ulcers (difficult-to-heal wounds).
In addition, the wound surface of the difficult-to-heal wound surface has complex cause, blood is often entrained and inflammatory reaction occurs, and the inflammatory reaction also needs energy supply, but the oxygen released by the oxygen carrier protein added into the dressing for promoting the wound healing can participate in the inflammatory reaction, so that the wound surface is promoted to heal quickly.
In addition, in the wound healing process, the growth environment of the skin can be changed, so that bacterial infection, especially anaerobic bacteria are generated in a large amount, and the oxygen-carrying protein added into the dressing for promoting wound healing has good curative effect on anaerobic bacteria infection, and can promote wound healing.
Preferably, in this embodiment, the mass ratio of the cell growth factor to the oxygen carrier protein is 1 to 20: 30-130.
The cell growth factor is used as a protein factor for promoting tissue healing, the cost is high, thousands of yuan is needed for a few micrograms, the cost of the oxygen-carrying protein is low, and the synergistic therapeutic effect of the cell growth factor and the oxygen-carrying protein can be realized on the premise of controlling the cost by controlling the proportion of the cell growth factor and the oxygen-carrying protein.
Specifically, in the present embodiment, the dressing for promoting wound healing is a liquid dressing.
The dressing for promoting wound healing is a liquid dressing, and small-dose local perfusion can not bring perfusion damage to the wound surface, but can relieve ischemia caused by wound blood vessels or ischemia caused by vascular occlusion caused by inflammation.
The dressing for promoting wound healing can be packaged in spray bottles for use, has the characteristics of no irritation, extremely low toxicity, quick response and long-term storage, can promote the repair and regeneration of tissue cells, reduce the probability of infection, accelerate the healing speed of wound surfaces, and simultaneously relieve the feeling of itching and discomfort.
The dressing for promoting wound healing provided by the invention has the advantages of small volume, easiness in carrying and convenience in use, is suitable for various scenes, can be used for self-treatment outside a hospital, and is easily sprayed on a wound to promote wound healing.
Preferably, in this embodiment, the dressing is a liquid dressing, and the pH of the dressing is 6.8 to 7.9.
Preferably, in this embodiment, the dressing is a liquid dressing, which is administered in the form of a spray.
In this embodiment, the dressing is a liquid dressing, and the solvent of the liquid dressing is water or glycerin.
It should be noted that the water mentioned in the present invention generally refers to water for injection, sterilized water for injection or water for injection for cell culture; the glycerin mentioned in the present invention generally refers to glycerin for injection.
In this embodiment, the dressing is a liquid dressing, which can be metered through the liquid dressing and dispensed into vials after being sterilized and filtered and/or terminally sterilized to form a sterile solution.
In the invention, the concentration of the cell growth factor comprehensively considers the cost and the concentration of the oxygen carrier protein, and the concentration of the oxygen carrier protein is determined according to the acting and oxygen carrying amount of the oxygen carrier protein.
Specifically, in the liquid dressing of the embodiment, the concentration of the cell growth factor is 10mg/L to 200mg/L, and the concentration of the oxygen carrier protein is 300mg/L to 1300mg/L.
Preferably, in the present embodiment, the cell growth factor is at least one selected from the group consisting of an epidermal growth factor, a fibroblast growth factor, a keratinocyte growth factor, a vascular endothelial growth factor, a transforming growth factor and a platelet-derived growth factor.
Compared with erythrocytes carrying oxygen in the same way, the oxygen carrying protein is a nanoscale protein, has the advantages of no need of matching, wide working temperature range (2-40 ℃), no coagulation risk, stability of 36 months and the like, and has the oxygen carrying capacity 7-9 times that of erythrocytes.
Preferably, in the present embodiment, the oxygen carrying protein is selected from at least one of hemoglobin, modified hemoglobin, myoglobin, and modified myoglobin.
Specifically, the modifier is at least one selected from glutaraldehyde, glyoxal, bis-aspirin, polyethylene glycol, 1, 5-hexadiene, dextran, starch and serum albumin.
The principle of modification of aldehydes and oxygen-carrying proteins is that aldehyde groups react with hanging amino groups on a protein main chain through Schiff base, and the principle of modification of the di-aspirin, polyethylene glycol, 1, 5-hexadiene, glucan, starch and the oxygen-carrying proteins is that carboxyl groups, hydroxyl groups or olefinic bonds carried by the substances are preactivated to react with the hanging amino groups on the protein main chain through substitution, and serum proteins can react with the amino groups hanging on the protein main chain through preactivation, wherein the activated products contain carboxyl groups. The modified oxygen-carrying protein has greatly reduced immunogenicity, high molecular weight, no permeation through kidney semipermeable membrane and small nephrotoxic effect.
Specifically, the modified hemoglobin can be prepared by the following operations: extracting red blood cells from whole blood of an animal; fully mixing red blood cells with the diluent to completely crack the red blood cells to obtain hemoglobin lysate; percolating, purifying and degerming the hemoglobin lysate to obtain a hemoglobin solution; mixing the hemoglobin solution with a protein modifier for reaction to obtain modified hemoglobin.
Specifically, the modified myoglobin can be prepared by the following operations: extracting skeletal muscle from skeletal muscle to obtain skeletal muscle extract; percolating, purifying and sterilizing the skeletal muscle extract to obtain myoglobin solution; and mixing the myoglobin solution with a protein modifier for reaction to obtain the modified myoglobin.
Wherein the extraction of skeletal muscle extract from skeletal muscle can be performed by techniques conventional in the art.
More preferably, the modifying agent is at least one selected from glutaraldehyde, glyoxal and dextran in the modified hemoglobin and the modified myoglobin.
It should be noted that hemoglobin and myoglobin can be extracted from natural tissues or synthesized artificially.
Specifically, hemoglobin can be prepared by the following operations: separating red blood cells from animal blood; mixing red blood cells with the diluent to crack the red blood cells to obtain a cracking solution; purifying and degerming the lysate to obtain the hemoglobin solution.
The diluent may be purified water, physiological saline or water for injection.
Preferably, the weight ratio of red blood cells to diluent may be 1:1.
specifically, the operation of removing viruses by filtration after purifying the lysate may be: the lysate is purified by diafiltration or chromatography, followed by filtration to remove bacteria.
Preferably, the content of the oxo-oxygen carrier protein in the oxygen carrier protein is less than or equal to 60 percent.
More preferably, the content of the oxo-oxygen carrier protein in the oxygen carrier protein is less than or equal to 10 percent.
In particular, the content of the oxidized oxygen carrier protein in the oxygen carrier protein solution can be made to be less than 60% (more preferably less than 10%) by introducing an inert gas into the oxygen carrier protein solution, wherein the inert gas is carbon dioxide, carbon monoxide or nitrogen.
The animal blood may be mammalian blood, preferably human blood, pig blood, bovine blood or horse blood.
Preferably, the dressing of the present embodiment further comprises an auxiliary material.
The auxiliary material accounts for 5-35% of the dressing by weight.
Specifically, the auxiliary materials are at least one selected from electrolytes, binders, antioxidants, analgesics, osmotic pressure regulators, stabilizers, bacteriostats, pH regulators, flocculants and film forming agents.
Preferably, the electrolyte is an inorganic salt. Specifically, the electrolyte contains at least one of sodium ion, potassium ion, magnesium ion, calcium ion, chloride ion, lactate ion, and bicarbonate ion.
Preferably, the binder is at least one selected from hydroxypropyl methylcellulose, polyethylene glycol, ethylcellulose, polyvinyl alcohol, starch slurry, chitosan, glycerol, glucose and gelatin slurry.
More preferably, the binder is selected from at least one of polyethylene glycol, starch slurry, chitosan, glycerol or glucose.
Preferably, the antioxidant is selected from at least one of sodium sulfite, sodium bisulfite, sodium metabisulfite, sodium thiosulfate, methionine, thioglycerol, cysteine, sodium borohydride and vitamin C.
More preferably, the antioxidant is selected from at least one of sodium thiosulfate, methionine, cysteine and vitamin C.
Preferably, the analgesic is selected from at least one of ibuprofen, indomethacin, tramadol, codeine, lidocaine, tramadol, fentanyl and sodium diclofenac.
More preferably, the analgesic is at least one selected from the group consisting of ibuprofen, indomethacin, codeine, lidocaine, fentanyl and sodium diclofenac.
Preferably, the osmolality adjusting agent is selected from at least one of sodium chloride, boric acid, glucose, borax, phosphate and citrate.
More preferably, the osmolality adjusting agent is selected from at least one of sodium chloride and glucose.
Preferably, the stabilizer is selected from at least one of polyethylene glycol, 2-hydroxypropyl-beta-cyclodextrin, glycerol, albumin, tryptophan, histidine, serine, threonine, glutamic acid and cysteine.
More preferably, the stabilizer is selected from at least one of polyethylene glycol, 2-hydroxypropyl-beta-cyclodextrin, glycerol, albumin and histidine.
Preferably, the bacteriostatic agent is at least one selected from benzalkonium chloride, benzalkonium bromide, benzyl alcohol, benzoic acid, lactic acid, chlorobutanol, hydroxymethyl ester, hydroxyethyl ester, methylparaben, chlorhexidine, ethylparaben, propylparaben, glycerin, peppermint oil and sorbic acid.
More preferably, the bacteriostatic agent is at least one selected from benzoic acid, benzyl alcohol, lactic acid, glycerin and peppermint oil.
Preferably, the pH adjuster is selected from sodium citrate, sodium carbonate, sodium bicarbonate, sodium hydroxide, sodium acetate or aqueous ammonia, preferably at least one of sodium carbonate, sodium bicarbonate and sodium hydroxide.
More preferably, the pH adjustor is selected from at least one of sodium carbonate, sodium bicarbonate and sodium hydroxide.
Preferably, the flocculant is selected from at least one of polymeric ferric chloride, liquid polymeric ferric sulfate, ferric chloride, chitosan, starch, cellulose, lignin, tannin, polyacrylamide and sodium polyacrylate.
More preferably, the flocculant is selected from at least one of ferric chloride, chitosan, starch, cellulose, polyacrylamide and sodium polyacrylate.
Preferably, the film forming agent is at least one selected from povidone K30, sodium carboxymethyl starch, polyvinyl alcohol, gum arabic powder and hydroxypropyl cellulose.
More preferably, the film forming agent is at least one selected from povidone K30, sodium carboxymethyl starch and polyvinyl alcohol.
The following are specific examples.
The basic fibroblast growth factor used in the examples is a commercially available product, grade pharmaceutical grade.
Example 1
1) Preparation of bovine hemoglobin
A healthy and qualified source of Ruxi yellow cattle (Qingdao Ruwei cattle industry Co., ltd.) was selected, and whole blood of the cattle was collected from the carotid artery puncture side with a sterile blood taking needle. 1000mL of the collected whole bovine blood was transferred to a 3L flexible bag, and a sodium citrate solution pH7.4 was added to make the mass concentration of sodium citrate 2.63wt%.
The whole blood of cattle after anticoagulation of sodium citrate passes through a hollow fiber membrane of 100kDa at a flow rate of 500mL/min, the diafiltration liquid is discarded, the separated red blood cells are collected, the filtered liquid passes through a filtering membrane of 0.4 mu m, the filtered liquid passes through the hollow fiber membrane of 0.2 mu m at a flow rate of 1L/min, the diafiltration product is collected, and the equal volume of water for injection is added under stirring to obtain the bovine hemoglobin.
Bovine hemoglobin was pumped into the 30kDa hollow fiber membrane at a rate of 200mL/min and the permeate was discarded until the bovine hemoglobin concentration in the permeate was less than 0.5mg/mL.
2) Purifying: the bovine hemoglobin solution after the above diafiltration was filtered through a 10kD membrane and then passed through an anion chromatography column system using 3g/L Tris buffer, the eluate was collected and deoxygenated continuously through a 0.22 μm filter until the dissolved oxygen number was below 0.05mg/mL. Obtaining the purified bovine hemoglobin.
3) Polymerization: under the condition of introducing nitrogen, adding 20 g/L100 mL of purified bovine hemoglobin into a polymerization kettle, controlling the pressure in the kettle to be within 0.3Mpa, adding 40mL of glutaraldehyde solution with the pressure of 3g/L, heating to 42 ℃ for reaction for 30min, regulating the pH to 8-10 by using borate buffer solution, adding 300mL of sodium borohydride solution for reaction for 30min, and removing small molecules from the product in the kettle through a 30kD filter membrane. Obtaining glutaraldehyde polymeric bovine hemoglobin, and then filtering and sterilizing the bovine hemoglobin by a 0.22 mu m filter membrane.
4) Preparation of liquid dressing
11.3g of glutaraldehyde polymeric bovine hemoglobin is weighed and placed in a sterile aluminum-plastic composite bag to be dissolved in 100mL of water for injection to obtain glutaraldehyde polymeric bovine hemoglobin solution, and 10mg of basic fibroblast growth factor is weighed and dissolved in 100mL of water for injection. The two were mixed, then 2.5g of sodium chloride, 180mg of cysteine, 40mg of sodium hydroxide were added respectively, and dissolved with stirring, and then 700mL of water for injection was added. And (3) sterilizing, filtering and deoxidizing the feed liquid in the aluminum-plastic composite bag through a 0.22 mu m filter membrane at the speed of 1L/min, and then subpackaging the feed liquid in aluminum metal spray bottles to obtain the liquid dressing.
Comparative example 1
1) Preparation of bovine hemoglobin
A healthy and qualified source of Ruxi yellow cattle (Qingdao Ruwei cattle industry Co., ltd.) was selected, and whole blood of the cattle was collected from the carotid artery puncture side with a sterile blood taking needle. 1000mL of the collected whole bovine blood was transferred to a 3L flexible bag, and a sodium citrate solution pH7.4 was added to make the mass concentration of sodium citrate 2.63%.
The whole blood of cattle after anticoagulation of sodium citrate passes through a hollow fiber membrane of 100kDa at a flow rate of 500mL/min, the diafiltration liquid is discarded, the separated red blood cells are collected, the filtered liquid passes through a filtering membrane of 0.4 mu m, the filtered liquid passes through the hollow fiber membrane of 0.2 mu m at a flow rate of 1L/min, the diafiltration product is collected, and the equal volume of water for injection is added under stirring to obtain the bovine hemoglobin.
Bovine hemoglobin was pumped into the 30kDa hollow fiber membrane at a rate of 200mL/min and the permeate was discarded until the bovine hemoglobin concentration in the permeate was less than 0.5mg/mL.
2) Purifying: the bovine hemoglobin solution after the above diafiltration was filtered through a 10kD membrane and then passed through an anion chromatography column system using 3g/L Tris buffer, the eluate was collected and deoxygenated continuously through a 0.22 μm filter until the dissolved oxygen number was below 0.05mg/mL. Obtaining the purified bovine hemoglobin.
3) Polymerization: under the condition of introducing nitrogen, adding 20 g/L100 mL of purified bovine hemoglobin into a polymerization kettle, controlling the pressure in the kettle to be within 0.3Mpa, adding 40mL of glutaraldehyde solution with the pressure of 3g/L, heating to 42 ℃ for reaction for 30min, regulating the pH to 8-10 by using borate buffer solution, adding 300mL of sodium borohydride solution for reaction for 30min, and removing small molecules from the product in the kettle through a 30kD filter membrane. Obtaining glutaraldehyde polymeric bovine hemoglobin, and then filtering and sterilizing the bovine hemoglobin by a 0.22 mu m filter membrane.
4) Preparation of liquid dressing
Weighing 11g of glutaraldehyde polymeric bovine hemoglobin, placing into a sterile aluminum-plastic composite bag, dissolving in 100mL of water for injection to obtain glutaraldehyde polymeric bovine hemoglobin solution, respectively weighing 2.5g of sodium chloride, 180mg of cysteine and 40mg of sodium hydroxide, adding into the glutaraldehyde polymeric bovine hemoglobin solution, stirring for dissolution, and then adding 700mL of water for injection. And (3) sterilizing, filtering and deoxidizing the feed liquid in the aluminum-plastic composite bag through a 0.22 mu m filter membrane at the speed of 1L/min, and then subpackaging the feed liquid in aluminum metal spray bottles to obtain the liquid dressing.
EXAMPLE 2 treatment of diabetic refractory skin ulcers rats
Test animals: 16 male rats with SD diabetes (. Gtoreq.8 weeks).
1. Experimental method
1.1 Grouping
The 16 experimental rats are randomly divided into a test group and a control group according to the body weight, wherein the dressing of the test group contains 8 cell growth factors.
1.2 Modeling
Diabetic foot ulcers are a serious complication of diabetes mellitus, and are a chronic wound that is difficult to heal due to foot infection, ulcers or deep tissue destruction caused by nerve abnormalities at the distal end of the lower limb, vascular lesions. Modeling here was aimed at creating a chronic wound model of diabetic foot ulcers with rats.
SD diabetic rats were all ear-marked.
Rats were induced with high fat diet for 6 weeks and developed insulin resistance. Then, 2% citric acid solution of streptozotocin is injected into the abdominal cavity, the injection dosage is 15-30 mg/kg, the complete excision is performed on the back skin of the rat, and the diabetic foot ulcer/wound surface model is induced to be manufactured.
1.3 Treatment of
Each group of experimental rats was treated following the protocol shown in table 1.
Table 1 test group
1.3 Evaluation of
The size of the wound at the site of the back resection was visually observed, and the size of the ulcer site was measured.
2. Results
The specific test results are shown in table 2 below.
Table 2 test results
As can be seen from Table 2, both the control and test groups can achieve promotion of healing of the skin of the ulcers in rats, but the test group promoted healing of the skin of the ulcers in rats significantly faster than the control group.
This shows that the basic fibroblast growth factor has obvious improvement effect on skin ulcer, but the liquid dressing of the basic fibroblast growth factor and glutaraldehyde polymeric bovine hemoglobin can more effectively accelerate the healing of skin ulcer under the synergistic cooperation of the basic fibroblast growth factor and glutaraldehyde polymeric bovine hemoglobin.
The above examples illustrate only a few embodiments of the invention, which are described in detail and are not to be construed as limiting the scope of the claims. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.

Claims (10)

1. A dressing for promoting wound healing comprising a cell growth factor and an oxygen carrier.
2. The dressing of claim 1, wherein the mass ratio of the cell growth factor to the oxygen carrier protein is 1-20: 30-130.
3. The dressing of claim 2, wherein the dressing is a liquid dressing;
the concentration of the cell growth factor is 10 mg/L-200 mg/L, and the concentration of the oxygen carrier protein is 300 mg/L-1300 mg/L.
4. The dressing according to any one of claims 1 to 3, wherein the cell growth factor is at least one selected from the group consisting of an epidermal growth factor, a fibroblast growth factor, a keratinocyte growth factor, a vascular endothelial growth factor, a transforming growth factor and a platelet-derived growth factor.
5. The dressing of claim 4 wherein the oxygen carrying protein is selected from at least one of hemoglobin, modified hemoglobin, myoglobin, and modified myoglobin.
6. The dressing of claim 5, wherein the modified hemoglobin is prepared by:
extracting red blood cells from whole blood of an animal;
fully mixing the red blood cells with a diluent to completely lyse the red blood cells, thereby obtaining a hemoglobin lysate;
percolating, purifying and degerming the hemoglobin lysate to obtain a hemoglobin solution;
mixing the hemoglobin solution with a protein modifier for reaction to obtain the modified hemoglobin;
alternatively, the modified myoglobin is prepared by the following operations:
extracting skeletal muscle from skeletal muscle to obtain skeletal muscle extract;
percolating, purifying and sterilizing the skeletal muscle extract to obtain myoglobin solution;
and mixing and reacting the myoglobin solution with a protein modifier to obtain the modified myoglobin.
7. The dressing of claim 6, wherein the protein modifier is selected from at least one of glutaraldehyde, glyoxal, bis-aspirin, polyethylene glycol, 1, 5-hexadiene, dextran, starch slurry, or serum albumin.
8. The dressing of claim 4 wherein the dressing further comprises an adjunct;
the auxiliary materials are selected from at least one of electrolyte, adhesive, antioxidant, analgesic, osmotic pressure regulator, stabilizer, bacteriostat, pH value regulator, flocculating agent and film forming agent.
9. The dressing of claim 5, wherein the oxygen carrier protein comprises less than or equal to 60% of the oxygen carrier protein.
10. The dressing of claim 9, wherein the content of the oxygenated oxygen carrier in the oxygen carrier is less than or equal to 10%.
CN202311384551.8A 2023-10-25 2023-10-25 Dressing for promoting wound healing Pending CN117100905A (en)

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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1499978A (en) * 2001-02-28 2004-05-26 ض� Manufacture of hemoglobin-based oxygen carrier
CN101678079A (en) * 2006-11-28 2010-03-24 韩兀制药株式会社 Modified erythropoietin polypeptides and uses thereof for treatment
US20100278887A1 (en) * 2007-06-18 2010-11-04 Centre National De La Recherche Scientifique Cnrs Use of a Haemoglobin for the Preparation of Dressings and Resulting Dressings
CN108354951A (en) * 2017-11-21 2018-08-03 北京颐方生物科技有限公司 A kind of oxygen-enriched gelling agent, preparation method and application promoting Wound healing
CN108752468A (en) * 2011-07-23 2018-11-06 萨斯托米德股份有限公司 Wound spray
CN110840655A (en) * 2019-08-05 2020-02-28 广东泓志生物科技有限公司 Bioactive dressing based on autologous concentrated growth factors and preparation method thereof

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1499978A (en) * 2001-02-28 2004-05-26 ض� Manufacture of hemoglobin-based oxygen carrier
CN101678079A (en) * 2006-11-28 2010-03-24 韩兀制药株式会社 Modified erythropoietin polypeptides and uses thereof for treatment
US20100278887A1 (en) * 2007-06-18 2010-11-04 Centre National De La Recherche Scientifique Cnrs Use of a Haemoglobin for the Preparation of Dressings and Resulting Dressings
CN108752468A (en) * 2011-07-23 2018-11-06 萨斯托米德股份有限公司 Wound spray
CN108354951A (en) * 2017-11-21 2018-08-03 北京颐方生物科技有限公司 A kind of oxygen-enriched gelling agent, preparation method and application promoting Wound healing
CN110840655A (en) * 2019-08-05 2020-02-28 广东泓志生物科技有限公司 Bioactive dressing based on autologous concentrated growth factors and preparation method thereof

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