CN105920659A - Bacteriostatic hydrogel dressing for wound repair and preparation method thereof - Google Patents
Bacteriostatic hydrogel dressing for wound repair and preparation method thereof Download PDFInfo
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- CN105920659A CN105920659A CN201610329358.8A CN201610329358A CN105920659A CN 105920659 A CN105920659 A CN 105920659A CN 201610329358 A CN201610329358 A CN 201610329358A CN 105920659 A CN105920659 A CN 105920659A
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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
- A61L26/0019—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
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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
- A61L26/0057—Ingredients of undetermined constitution or reaction products thereof
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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
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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
- A61L26/008—Hydrogels or hydrocolloids
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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
- A61L26/009—Materials resorbable by the body
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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
- 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
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Abstract
The invention provides a bacteriostatic hydrogel dressing material for wound repair and a preparation method thereof. The bacteriostatic hydrogel dressing material provided by the invention is solid-state fine powder consisting of epsilon-polylysine, gamma-polyglutamic acid, acellularized scaffold powder and cross-linking agents; and the bacteriostatic hydrogel dressing can be directly spread on the wound surface or can be pasted onto the wound surface after absorbing certain water to become gel. According to composition ingredients, the mass ratio of the epsilon-polylysine to the gamma-polyglutamic acid is (100:1) to (1:10); the acellularized scaffold powder is 0.01 to 20 percent of the total mass of the epsilon-polylysine and the gamma-polyglutamic acid; and the cross-linking agents account for 0.01 percent to 40 percent of the weight of the hydrogel dry powder dressing. The wound repair material has good hygroscopicity; the water absorption capability reaches 200 times of the self weight; the wound repair material also has good bacteriostatic capability, so that the additional addition of antibiotics or silver-containing bacteriostatic ingredients is not needed; and meanwhile, good mechanical performance is also realized. In addition, the bacteriostatic hydrogel dressing material has the advantages that the preparation method is simple; the repeatability is good; and the large-scale production can be easily realized.
Description
Technical field
The present invention relates to a kind of bacteriostatic water gel dressing for wound repair and preparation method, specifically by γ-many polyglutamics
The aquogel type wound repair material of acid rear preparation crosslinked with ε-poly-l-lysine complex and preparation method.
Background technology
The hemostasis being applied to clinic at present is mainly gelfoam and water miscible cellulose family with wound repair material.Water-setting
Glue is a kind of novel biomaterial, and it has important using value in terms of repair in trauma.Clinical practice hydrogel at present
Dressing is mainly composed of carboxymethyl cellulose and some other supplementary element, such as pectin, Xanthan gum, propyleneglycoles etc..
Such dressing is provided that the moist environment needed for wound healing, beneficially cell proliferation, epithelial migration and granulation growth, energy
Pain relieving, more will not damage wound during change dressings, and can fill the deep dead space of wound, the glue in wound self transudate
Former protein degradation enzyme decomposes downright bad material, to reach the effect of autolytic debridement.Such aerogel dressing is a large amount of at absorption wound surface
After exudate, the expansion of colloid and cause dressing and wound separation, make antibacterial amount reproduction, easily cause traumatic infection.
In processes of wound repair, traumatic infection and the complication that causes are the weights faced in the current clinical practice of Hydrogels dressing
Want problem.
ε-the poly-D-lysine of heterogeneous, by fermentable, is generally made up of 25-30 bad amino acid residue
Polycation polymerized peptides, such polyamino acid material has good water solublity, and side chain contains substantial amounts of amino, can be thin
Cellular surface negative charge effect, has superpower cell adhesion ability.ε-poly-D-lysine antimicrobial spectrum is wide, in acid and subacidity
Environment all there is certain fungistatic effect to gram positive bacteria, gram negative bacteria, yeast, mycete, especially to it
His natural antiseptic agent be difficult to suppression gram-negative escherichia coli, Salmonella fungistatic effect the best, be current sky
Preservative so has the microbiology class food preservative of superior antiseptic property and huge business potential.Chinese patent (patent
Application number: CN201410571048.8) disclose the system of a kind of epsilon-polylysine-vitamin e succinate amide complex
Standby and be used as the application in terms of food fresh keeping, improve the fat-soluble of epsilon-polylysine, improve the fresh-keeping effect to meat product.
At present, yet there are no ε-poly-D-lysine is used for the antibacterial aspect of wound surface document report.
γ-polyglutamic acid (γ-PGA) is that one is synthesized by multiple bacillus and is secreted into extracellular water soluble polymer amount
Amino acid polymer, is polymerized by gamma-glutamyl key by D-Glu and Pidolidone.γ-polyglutamic acid is hydrophilic
Polymer network, there is high-hydroscopicity, higher mechanical strength, controlled degradation and biocompatibility, be a kind of system
Standby wound dressing and the ideal material of tissue engineering bracket.But because of the water solublity that it is strong, it is difficult to form gel, such material
Separately as Wound dressing, it is difficult to be formed stable gel state after absorbing wound fluid, thus impact is stopped blooding and repairs
Effect.Wang Jing etc. report with γ-polyglutamic acid as raw material, and allyl bromide, bromoallylene and bromination of n-butane make esterifying agent, prepare ester
Changing γ-polyglutamic acid, continue to be esterified γ-polyglutamic acid as monomer, azodiisobutyronitrile makees initiator, methacrylic acid
Hydroxyl ethyl ester makees cross-linking agent, and preparation γ-polyglutamic acid hydrogel (China's Tissue Engineering Study and clinical rehabilitation, 2008 years the
Volume 12 the 1st phase).This gel shows obvious absorbability, such as γ-polyglutamic acid hydrogel that esterification degree is 40%
Water absorption rate can reach 44 times of own wt, but the method prepares the synthesis technique that hydrogel needs are complicated, water-setting colloidality
Matter poor reproducibility.Chinese patent (Publication No. CN105268015A) discloses a kind of anti-bacterial hydrogel composite,
It is prepared by mixing into hydrogel by chitosan and gamma-polyglutamic acid-with certain mass proportioning, after by polyvinylpyrrolidone
(PVP) nanometer silver loaded is dispersed in above-mentioned hydrogel and is prepared from.This patent use nanometer silver as antimicrobial component,
Though anti-microbial property is relatively strong, but has serious zest, poor biocompatibility to wound surface, it is impossible to degraded, environment is caused
Pollute.
Acellularized valve is derived from the extracellular matrix after organism normal structure, de-cell, and it is mainly by structural protein
Form with functional protein, the multiple somatomedin the most also comprised, in tissue repair and reconstruction, have important facilitation.
The biologic bracket material of Acellularized valve composition is widely used in the regenerative medicine neck of surgical reconstruction and tissue and organ
Territory such as cardiac valve, blood vessel, skin, nerve, tendon and small intestinal submucosa etc..
The present invention will have the cation ε-poly-D-lysine of broad-spectrum antibacterial effect, remove cell in conjunction with can promote wound healing
Support powder, is first combined through electrostatic with anion γ-polyglutamic acid of strong water absorbing capacity and prepares complex, continue with low concentration
Cross-linking agents complex, preparation has the hydrogel of certain mechanical performance, by washing, is dried, pulverizes and be prepared into
To having the medical aquogel powder dressing of bacteriostasis, can directly be sprinkling upon wound surface or absorb certain water in advance and become gel
Stick on wound surface, wound healing.
Summary of the invention
It is desirable to provide one is without outer added with antibiotic or silver class antipathogenic composition, can be used for wound repair self has
The aerogel dressing of bacteriostasis.This dressing can directly be sprinkling upon wound surface or absorb certain water in advance and become gel to stick on wound
Face.The present invention will have the cation ε-poly-D-lysine of broad-spectrum antibacterial effect, remove cell in conjunction with can promote wound healing
Support composition, is first combined through electrostatic with anion γ-polyglutamic acid of strong water absorbing capacity and prepares complex, continue with low concentration
Cross-linking agents complex, preparation has the hydrogel of certain mechanical performance, by washing, is dried, pulverizes and be prepared into
To the medical aquogel powder dressing with bacteriostasis.
The present invention can be realized by following technical proposal:
The bacteriostatic water gel dressing of a kind of wound repair is by ε-poly-D-lysine, γ-polyglutamic acid, Acellularized valve powder
End and the solid fines of cross-linking agent composition.
The bacteriostatic water gel solids fine powder dressing of above-mentioned wound repair, it can cross the 40 medicinal sieves of mesh.
Above-mentioned ε-poly-D-lysine is 100: 1~1: 10 with the ratio of the quality of γ-polyglutamic acid, and the ratio of preferred mass is
100: 1~1: 1;Acellularized valve powder accounts for the 0.01~20% of the ε-poly-D-lysine gross mass with γ-polyglutamic acid, preferably
0.1~15%;Cross-linking agent accounts for the 0.01%~30% of hydrogel dry powder dressing weight, preferably 0.01%~20%.
Above-mentioned ε-poly-D-lysine is selected from molecular weight 1000~6000Da ε-poly-D-lysine, and preferred molecular weight exists
ε-the poly-D-lysine of 2000~5000Da;γ-polyglutamic acid is selected from molecular weight at 500,000~1,200,000 Da γ-many polyglutamics
Acid, preferred molecular weight is at 700,000~1,000,000 Da γ-polyglutamic acid;Acellularized valve powder derives from the viscera tissue of pig;
Cross-linking agent is selected from paraformaldehyde, glutaraldehyde, 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (EDC), N, N-
One or more in DIC (DIC).
The preparation method of the bacteriostatic water gel dressing of above-mentioned wound repair, it is characterised in that: this preparation method include with
Lower step:
(1) weigh ε-poly-D-lysine to be dissolved in respectively in deionized water with certain concentration with γ-polyglutamic acid, preparation
Two phase liquid, equal-volume mixing, stir after, Acellularized valve powder is directly dispersing in pluralgel, stirring
Uniformly prepare complex gel;
(2) taking appropriate cross-linking agent to add in complex gel prepared by (1), reaction 3h is stirred at room temperature, preparation is stable
Cross-linked gel;
(3) cross-linked gel prepared by (2) is dispersed in the dehydrated alcohol of appropriate volume, stirring dehydration 30min, takes out
Filter, wash, dry, prepare gel lump, be ground into fine powder, prepare aerogel dressing powder.
The concentration that in above-mentioned steps (1), ε-poly-D-lysine is dissolved in deionized water is 0.1~50% (w/v), preferably 5~50%
(w/v);It is 0.01~50% (w/v) that γ-polyglutamic acid is dissolved in concentration in deionized water, preferably 1~40% (w/v).
Amount is system 0.01~15% (w/v) of addition cross-linking agent in above-mentioned steps (2), preferably 0.05~10% (w/v),
Being dissolved in a small amount of deionized water by cross-linking agent and add system, thus cross-linking agent mixes evenly in system, reacts more fast
Efficiently, prepared gel structure is tight, mechanical strength is more excellent for speed.
In above-mentioned steps (3), addition dehydrated alcohol volume is 5~100 times of gel rubber system volume, for cost-effective, excellent
Select 5~60 times.
The present invention compares similar aerogel dressing and there is following advantage: 1) dressing itself is containing natural antipathogenic composition ε-many
Polylysine, it is not necessary to add antibiotic or silver class antipathogenic composition, effectively reduce the stimulation to wound surface;2) dressing is by sky
So polyamino acid class material forms with Acellularized valve, has good biocompatibility, biodegradability, and degraded is produced
Thing has Nutrition to wound surface;3) comparing the dressing of Clinical practice, dressing materials prepared by the present invention has super-strong moisture absorbing
Ability.
Accompanying drawing explanation
Accompanying drawing 1 is the gel dressing powder SEM figure organizing 1 preparation in embodiment 4
Accompanying drawing 2 is the gel dressing powder SEM figure organizing 4 preparations in embodiment 4
Accompanying drawing 3 is the gel dressing powder SEM figure organizing 10 preparations in embodiment 4
Accompanying drawing 4 is the gel dressing powder SEM figure organizing 17 preparations in embodiment 4
Accompanying drawing 5 embodiment 4 is organized 1, organizes 4, group 10 and group 17 Young's modulus measurement results
Specific embodiment
Embodiment 1 water absorption rate measures
By prepared hydrogel pressed powder, weigh to obtain W0Value is placed in the weighing botle filling deionized water, the most swelling,
Blot surface moisture with filter paper, weigh to obtain WnValue.Sample water absorption rate calculates according to following equation: water absorption rate (%)=100 ×
(Wn-W0)/W0
Embodiment 2 bacteriostatic experiment
(1) preparation for examination bacteria suspension takes E.coli K88, strains of streptococcus is inoculated in LB culture medium,
Cultivate 24h for 37 DEG C, choose single bacterium in 3ml LB fluid medium with inoculating loop, 37 DEG C, 200r/min concussion cultivate system
Standby bacteria suspension.
(2) take 100 μ L bacteria suspensions and be spread evenly across LB/SS flat board, after placing about 30min, a diameter of 7mm
Wound dressing be placed on flat board, the drug susceptability test paper after antibacterials are soaked simultaneously is placed on flat board as positive control, steams
Reagent paper after distilled water is soaked is put as negative control, cultivates 24h, measures the diameter of each inhibition zone with slide gauge, often for 37 DEG C
Kind sample is repeated 3 times and takes its meansigma methods, observes the size of inhibition zone.This experiment uses 0.2g/L lincomycin and 0.5g/L
Piperacillin as positive control.
The preparation of embodiment 3 Acellularized valve powder
Weigh pig heart 100g, be cut into the fragment of 1-3cm, to the container filling 300mL 1% sodium lauryl sulphate
Shake (SDS) de-cell to process, processing procedure is changed fresh SDS treatment fluid often, until heart fragment becomes half
Transparent network structure, incline SDS treatment fluid, continues to remove SDS 5-8 time with 300mL distilled water flushing, prepares wet
Profit support, can be prepared Acellularized valve powder by following treatments, and (1) is freeze-dried, pulverization process prepares support
Powder;Or (2) moistening support is first dispersed in 0.1M hydrochloric acid, the pepsin digestion adding weight support frame 10% processes
48h, sodium hydroxide regulation pH to 7.4, obtained by freeze drying support powder.
The preparation of embodiment 4 bacteriostatic water gel dressing
According to table 1 prescription, weigh ε-poly-D-lysine and put in two 100ml beakers respectively with γ-polyglutamic acid, point
Jia Ru not be stirred at room temperature and make it be completely dissolved by 10ml distilled water, prepare two-phase polymer solution;By ε-Poly-L-Lysine Solution
It is slowly added to the γ being stirred continuously-polyglutamic acid solution, prepares complex gel;According to table 1 prescription, weigh carbon
Diimine class cross-linking agent is dissolved in 5mL distilled water or directly with pressed powder or be heated to be steam, is slowly added to or is passed through
State in complex gel, reaction 3h is stirred at room temperature, prepares stable cross-linked gel;By gel dispersion after above-mentioned crosslinking such as
In the dehydrated alcohol of volume shown in table 1, stirring dehydration 30min, sucking filtration, dehydrated alcohol 30ml wash three times, 80 DEG C
Dry, prepare gel lump, pulverize, cross the 40 medicinal sieves of mesh, prepare aerogel dressing powder.Prepared hydrogel
Dressing is according to embodiment 1, and embodiment 2 carries out water absorption rate experiment and bacteriostatic experiment, and result is summed up and table 1.
Embodiment 5 bacteriostatic water gel dressing surface morphologic observation
Taking bacteriostatic water gel solid state prepared by different prescription and be dried fine materials, be fixed on sample stage, surface metal spraying is placed on to be swept
Retouch electricity Microscopic observation.Hydrogel internal structure is carried out electron-microscope scanning result show: the ε-poly-D-lysine/γ-poly of different proportion
Glutamic acid, after the cross-linking agents of variable concentrations, the hydrogel material of formation has the structure of three-dimensional, porous, and accompanying drawing 1 is
Organizing the gel dressing SEM figure of 1 preparation in embodiment 3, accompanying drawing 2 is the gel dressing SEM figure organizing 4 preparations in embodiment 3,
Accompanying drawing 3 is the gel dressing SEM figure organizing 10 preparations in embodiment 3, and accompanying drawing 4 is that the gel organizing 17 preparations in embodiment 3 applies
Material SEM figure.
Embodiment 6 bacteriostatic water gel dressing mechanics properties testing
Take bacteriostatic water gel solid state prepared by different prescription and be dried fine materials, PBS soaks 24h, the thickness of measuring samples,
Sample is placed on the platform of biomaterial dynamic test machine, is compressed test according to the speed of 1mm/min.By answering
Force-strain curve calculates the fracture strength of sample, extension at break and Young's modulus (n=3).Embodiment 3 is organized 1, organizes 4, organize 10
With organize 17 Young's modulus measurement results such as accompanying drawing 5, be all higher than 2kPa, show that there is good mechanical strength.
Claims (8)
1. the bacteriostatic water gel dressing of a wound repair and preparation method, it is characterised in that: by γ-polyglutamic acid, ε-
Poly-D-lysine, Acellularized valve powder and the solid fines of cross-linking agent composition.
The bacteriostatic water gel dressing of wound repair the most according to claim 1, it is characterised in that: described solid fines
The 40 medicinal sieves of mesh can be crossed.
3. according to the bacteriostatic water gel dressing of the wound repair described in claim 1 or claim 2, it is characterised in that: institute
ε-the poly-D-lysine stated is 100: 1~1: 10 with the ratio of the quality of γ-polyglutamic acid, and Acellularized valve powder accounts for ε-poly and relies ammonia
The 0.01-20% of the gross mass of acid and γ-polyglutamic acid, cross-linking agent accounts for the 0.01%~40% of hydrogel dry powder dressing weight.
4. according to the bacteriostatic water gel dressing of the wound repair described in claim 1 or claim 2 or claim 3, its
It is characterised by: described ε-poly-D-lysine is selected from molecular weight 1000~6000Da ε-poly-D-lysine, and γ-polyglutamic acid is selected from
Molecular weight is at 500,000~1,200,000 Da γ-polyglutamic acid, and Acellularized valve powder derives from the viscera tissue of pig;Cross-linking agent is selected from
Paraformaldehyde, glutaraldehyde, 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride, N, N-DIC.
5., according to the preparation method of the bacteriostatic water gel dressing of the wound repair described in claim 1 or claim 2, it is special
Levy and be: this preparation method comprises the following steps:
(1) weigh ε-poly-D-lysine to be dissolved in respectively in deionized water with certain concentration with γ-polyglutamic acid, prepare biphase
Solution, equal-volume mixing, stir after, Acellularized valve powder is directly dispersing in pluralgel, stir preparation
Complex gel;
(2) take appropriate cross-linking agent to add in complex gel prepared by (1), reaction 3h is stirred at room temperature, prepares stable friendship
Connection gel;
(3) cross-linked gel prepared by (2) is dispersed in the dehydrated alcohol of appropriate volume, stirring dehydration 30min, sucking filtration,
Washing, drying, prepare gel lump, be ground into fine powder, prepare aerogel dressing powder.
The preparation method of the bacteriostatic water gel dressing of wound repair the most according to claim 5, it is characterised in that: described
The concentration that in step (1), ε-poly-D-lysine is dissolved in deionized water is 0.1~50% (w/v), γ-polyglutamic acid be dissolved in from
In sub-water, concentration is 0.01~50% (w/v).
The preparation method of the bacteriostatic water gel dressing of wound repair the most according to claim 5, it is characterised in that: described
Adding amount is system 0.01~15% (w/v) of cross-linking agent in step (2), the mode of addition has: 1. pressed powder directly adds
Enter, the most first cross-linking agent is dissolved in a small amount of deionized water and adds system, 3. cross-linking agent is flashed to steam and be passed through system.
The preparation method of the bacteriostatic water gel dressing of wound repair the most according to claim 5, it is characterised in that: described
In step (3), addition dehydrated alcohol volume is 5~100 times of gel rubber system volume.
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