CN107082907A - Preparation method of porous silk gum/polyvinyl alcohol gel and products thereof and application - Google Patents

Preparation method of porous silk gum/polyvinyl alcohol gel and products thereof and application Download PDF

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Publication number
CN107082907A
CN107082907A CN201710288164.2A CN201710288164A CN107082907A CN 107082907 A CN107082907 A CN 107082907A CN 201710288164 A CN201710288164 A CN 201710288164A CN 107082907 A CN107082907 A CN 107082907A
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China
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silk gum
polyvinyl alcohol
alcohol gel
porous silk
porous
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Inventor
何华伟
陶刚
王叶菁
蔡蕊
赵萍
夏庆友
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Southwest University
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Southwest University
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    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J9/00—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof
    • C08J9/28—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof by elimination of a liquid phase from a macromolecular composition or article, e.g. drying of coagulum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
    • 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
    • A61L26/0014—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials obtained by reactions only involving carbon-to-carbon unsaturated bonds
    • 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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    • 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
    • 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
    • A—HUMAN NECESSITIES
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    • 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
    • A61L26/0085—Porous materials, e.g. foams or sponges
    • C—CHEMISTRY; METALLURGY
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    • C08J3/00—Processes of treating or compounding macromolecular substances
    • C08J3/02—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques
    • C08J3/03—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media
    • C08J3/075—Macromolecular gels
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
    • C08L29/00—Compositions of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by an alcohol, ether, aldehydo, ketonic, acetal or ketal radical; Compositions of hydrolysed polymers of esters of unsaturated alcohols with saturated carboxylic acids; Compositions of derivatives of such polymers
    • C08L29/02—Homopolymers or copolymers of unsaturated alcohols
    • C08L29/04—Polyvinyl alcohol; Partially hydrolysed homopolymers or copolymers of esters of unsaturated alcohols with saturated carboxylic acids
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    • C08L89/00—Compositions of proteins; Compositions of derivatives thereof
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/60—Salicylic acid; Derivatives thereof
    • A61K31/612—Salicylic acid; Derivatives thereof having the hydroxy group in position 2 esterified, e.g. salicylsulfuric acid
    • A61K31/616—Salicylic acid; Derivatives thereof having the hydroxy group in position 2 esterified, e.g. salicylsulfuric acid by carboxylic acids, e.g. acetylsalicylic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/7036—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin having at least one amino group directly attached to the carbocyclic ring, e.g. streptomycin, gentamycin, amikacin, validamycin, fortimicins
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    • 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
    • A61L2300/21—Acids
    • 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
    • A61L2300/23—Carbohydrates
    • A61L2300/232—Monosaccharides, disaccharides, polysaccharides, lipopolysaccharides
    • 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
    • A61L2300/41—Anti-inflammatory agents, e.g. NSAIDs
    • C—CHEMISTRY; METALLURGY
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    • C08J2389/00—Characterised by the use of proteins; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2429/00—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by an alcohol, ether, aldehydo, ketonic, acetal, or ketal radical; Hydrolysed polymers of esters of unsaturated alcohols with saturated carboxylic acids; Derivatives of such polymer
    • C08J2429/02—Homopolymers or copolymers of unsaturated alcohols
    • C08J2429/04—Polyvinyl alcohol; Partially hydrolysed homopolymers or copolymers of esters of unsaturated alcohols with saturated carboxylic acids
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
    • C08L2203/00—Applications
    • C08L2203/02—Applications for biomedical use

Abstract

The present invention relates to a kind of preparation method of porous silk gum/polyvinyl alcohol gel and products thereof and application, preparation method is in addition mould after silk gum solution is mixed with poly-vinyl alcohol solution, in the case where temperature is less than 0 DEG C and higher than 0 DEG C, multigelation to aperture is formed after standing, then it is freeze-dried, obtain porous silk gum/polyvinyl alcohol gel, this method is resulted in the good porous silk gum/polyvinyl alcohol gel of preferable mechanical property and moisture pick-up properties, and do not cause the change of silk gum structure, there is good anti-microbial property after carrying medicament, no cytotoxicity, it can prevent bacterium from being infected to wound site, the method of the present invention is simple to operate, mild condition, environmental protection, porous silk gum/polyvinyl alcohol gel of the carrying medicament of preparation is expected to be used for the associated biomolecule medical material such as antiseptic dressing field.

Description

Preparation method of porous silk gum/polyvinyl alcohol gel and products thereof and application
Technical field
The invention belongs to technical field of biological material, it is related to the preparation method of porous silk gum/polyvinyl alcohol gel, further relates to by this Product made from method and application.
Background technology
Infection is one of wound care area the most universal complication, and it is several that severe open trauma patient infects Rate is up to more than 10, how to be effectively reduced the risk that open wound infection occurs, has become wound repair neck Domain focus and difficulties urgently to be resolved hurrily.The application of novel antibacterial material is expected to solve this problem, uses suitable part Antiseptic dressing can fully reduce the infection of wound, while promoting wound healing comprehensively.
In recent years, developing rapidly with materialogy and organizational engineering, many Medical dressings arise at the historic moment.It is preferable Dressing should have absorbable wound fluid, keep the temperature and humidity, good gas permeability, antibacterial of wound facing face disappear Scorching the advantages of.The requirement that aerogel dressing is closer to preferable dressing has the advantages that high-selenium corn, moisturizing, easily changed.Because water Gel is to be crosslinked obtained one kind by covalent bond, ionic bond or hydrogen bond etc. by hydrophilic macromolecular compounds to contain a large amount of water Point, the swelling body with tridimensional network, possess certain compression strength, and the environment moistened can be provided for wound, with There is preferable compatibility in biological tissue.Exactly because so high water content, allows hydrogel arbitrarily moulding, is suitable for any The surface of a wound at position, necessary condition is provided for the healing of wound, and its biocompatibility is also better than fiber, film class dressing.Thus The biomedical engineering fields such as wound dressing are greatly paid close attention to, and have extremely good application prospect.
But the raw material of traditional macromolecule hydrogel is generally commercial synthesis, preparation cost is higher and preparation link is cumbersome multiple It is miscellaneous, the reasons such as environmental pollution easily are brought in preparation process, water absorbent gel is prepared increasingly by everybody with natural polymer Favor.Silk gum is wrapped around a kind of natural macromolecular viscous protein on fibroin fiber top layer, and protection is played to fibroin and is adhered Effect, be one of the two kinds of main albumen for constituting silk cocoon, account for the 25% of silk cocoon.In 18 kinds of amino acid of composition silk gum It is the necessary amino acid of human body to have 8 kinds, glycine, serine, alanine and tyrosine containing high content, has many pharmacology Effect, healthcare function and very high nutritive value.It is such as inoxidizability, anti-in addition, sericin has a good bioactivity Bacterium, anti-crosslinking, the propagation for promoting cell and the function such as differentiation, and sericin has biological degradability, hydrophily and gathers around There is abundant polar group, therefore receive more and more attention and apply to regenerative medicine field.But pure silk gum water-setting Colloidality matter is fragile, and easily turns into the hotbed of bacteria breed in a humid environment.Therefore research and development with good biocompatibility, Good mechanical property and the silk gum anti-biotic material of stability, give full play to the advantage of sericin, with important economy effect Benefit and social effect.
The content of the invention
In view of this, it is an object of the invention to provide a kind of preparation method of porous silk gum/polyvinyl alcohol gel；This hair The bright second purpose is to provide the product as made from the above method；The third object of the present invention is to provide using described porous Silk gum/polyvinyl alcohol gel prepares porous silk gum/polyvinyl alcohol gel of carrying medicament；The fourth object of the present invention is to provide The preparation method of porous silk gum/polyvinyl alcohol gel of carrying medicament；The fifth object of the present invention is to provide the load medicine Application of the porous silk gum/polyvinyl alcohol gel of thing in antiseptic dressing is prepared.
To reach above-mentioned purpose, the present invention provides following technical scheme：
1st, the preparation method of porous silk gum/polyvinyl alcohol gel, comprises the following steps：Silk gum solution and polyvinyl alcohol is molten Added after liquid mixing in mould, multigelation to aperture is formed at less than 0 DEG C and higher than 0 DEG C after standing, is then freezed Dry, obtain porous silk gum/polyvinyl alcohol gel.
Simple to operate, mild condition of the invention, environmental protection, preparing porous silk gum/polyvinyl alcohol gel has good power Learn performance, moisture pick-up properties.
In the present invention, the mass fraction of the silk gum solution is 1~5%；The mass fraction of the poly-vinyl alcohol solution is 1 ~5%.
It is preferred that, the volume ratio of the silk gum solution and poly-vinyl alcohol solution is 1：1~4：1.
It is furthermore preferred that the multigelation be at -20 DEG C and 18~25 DEG C alternately；Or in -80 DEG C and 18~25 At DEG C alternately；Or at -196 DEG C and 18~25 DEG C alternately.Different temperature obtains different aperture size and hole The gel of rate, therefore controllable pore size and porosity can be obtained by regulating and controlling temperature.
The present invention prepares silk gum powder by HTHP and freeze-drying, is specially：Silk cocoon is cut into fractionlet, in 120 DEG C, handle under the conditions of 0.1Mpa, then with filtered through gauze, remove insoluble substance, freeze-drying obtains silk gum powder；So Molten obtained silk gum solution is weighed afterwards.
2nd, porous silk gum/polyvinyl alcohol gel as made from the preparation method.
3rd, porous silk gum/polyvinyl alcohol gel of carrying medicament is prepared using the porous silk gum/polyvinyl alcohol gel.
In the present invention, the medicine is gentamicin and aspirin, and gentamicin and Aspirin concentrations are respectively 0.3 ~0.5mg/ml and 2~5mg/ml.
4th, the preparation method of porous silk gum/polyvinyl alcohol gel of the carrying medicament, by the porous silk gum/polyethylene Alcogel is infiltrated in the solution containing medicine, obtains porous silk gum/polyvinyl alcohol gel of carrying medicament.
5th, application of the porous silk gum/polyvinyl alcohol gel of the carrying medicament in antiseptic dressing is prepared.
The present invention prepares silk gum powder by HTHP and freeze-drying, sericin solution is blended with PVA molten Liquid prepares silk gum/polyvinyl alcohol gel with preferable mechanical property by the method for multigelation.After freeze-drying Silk gum/polyvinyl alcohol gel there is controllable pore size and porosity.Medicine (gentamicin on being loaded on porous gel And aspirin)
Make it have effect of good anti-microbial property and anti-inflammatory analgesic.Simple to operate, mild condition of the invention, green ring Protect, silk gum/polyvinyl alcohol gel of the medicine loading of preparation has the anti-inflammatory effect of good mechanical property, moisture pick-up properties and antibacterial It is expected to be used for the associated biomolecule medical material such as antiseptic dressing field.
The beneficial effects of the present invention are：The present invention is by by sericin solution, by repeatedly after being blended with PVA solution The method of freeze thawing prepares silk gum/polyvinyl alcohol gel with preferable mechanical property.Silk gum after freeze-drying/poly- Ethene alcogel has controllable pore size and porosity.Medicine (gentamicin and A Si on being loaded on porous gel Woods) make it have effect of good anti-microbial property and anti-inflammatory analgesic.SEM test results show silk gum/polyvinyl alcohol gel tool There is the small gap of controllable macropore；FT-IR, XRD, TGA and fluorometric investigation show that silk gum and PVA are well mixed and do not cause silk gum knot The change of structure；Contact angle test shows that silk gum/polyvinyl alcohol gel has good moisture pick-up properties；Antibacterial test shows to load medicine Silk gum/PVA of thing has good anti-microbial property；Cytotoxicity experiment result shows silk gum/polyvinyl alcohol gel and loading Silk gum/polyvinyl alcohol gel of medicine does not have cytotoxicity；Simulation wound infection model measurement shows silk gum that medicine loads/poly- Ethene alcogel can protect wound to prevent bacterium from being infected to wound site.Simple to operate, mild condition of the invention, green ring Protect, silk gum/polyvinyl alcohol gel that the medicine of preparation is loaded is expected to be used for the associated biomolecule medical material such as antiseptic dressing field.
Brief description of the drawings
In order that the purpose of the present invention, technical scheme and beneficial effect are clearer, the present invention provides drawings described below and carried out Explanation：
Fig. 1 detects the hole situation for silk gum/polyvinyl alcohol gel that the different proportion prepared under different freezing conditions is put： (A) SEM observes the hole of the silk gum/polyvinyl alcohol gel prepared under different freezing conditions；(B) prepared under different freezing conditions The pore size statistics of different proportion silk gum/polyvinyl alcohol gel；(C) silk gum/polyvinyl alcohol prepared under -80 DEG C of freezing conditions The porosity of gel (4/1-4/2-4/4)；
The sign of Fig. 2 silk gum/polyvinyl alcohol gel；(A) FT-IR detections show that silk gum is well mixed with PVA；(B) mechanics Performance detection adds the mechanical property that PVA significantly improves silk gum；(C) XRD detections, which add PVA, does not influence the crystalline structure of silk gum； (D) TGA detections show that PVA can improve stability；
Fig. 3 silk gum/polyvinyl alcohol gel water imbibition test；(A) silk gum/PVA contact angles represent quick absorption；(B) silk gum/ The PVA swelling equilibrium times show that silk gum is the main matter quickly absorbed water；(C) silk gum/PVA water absorption rates show in acid or alkali environment Can largely it absorb water；
Fig. 4 silk gum/polyvinyl alcohol gel fluoroscopic examination shows that silk gum forms mutually perforation gel with PVA；
The silk gum that Fig. 5 medicines are loaded/polyvinyl alcohol gel insoluble drug release detection：(A) standard curve of gentamicin, (B) The standard curve of aspirin, the release profiles of (C) gentamicin, the release profiles of (D) aspirin；
The Bactericidal curves test for silk gum/polyvinyl alcohol gel that Fig. 6 medicines are loaded and Bactericidal test：(A) (C) suppresses big Enterobacteria, (B) (D) suppress staphylococcus aureus；
Silk gum/polyvinyl alcohol gel cytotoxicity test that Fig. 7 medicines are loaded：(A) cell viability of NIH3T3 cells is surveyed Examination, the cell viability test of (B) HEK293 cells, the life or death cell dyeing detection of (C) NIH3T3 cells, (D) HEK293 cells Life or death cell dyeing detection；
The wound infection model inspection for silk gum/polyvinyl alcohol gel that Fig. 8 medicines are loaded：(A) wound infection model, (B) are The activity statistics of cell and bacterium, (C) are cell and bacterium in HEK293 cell wound models in NIH3T3 cell wound models Activity statistics, (D) be that life or death cell dyeing is detected in NIH3T3 cell wound models, (E) is NIH3T3 cell wound models Middle bacterium coated plate detection vigor, (F) be that life or death cell dyeing is detected, (G) is HEK293 cells in HEK293 cell wound models Bacterium coated plate detects vigor in wound model.
Embodiment
Below in conjunction with accompanying drawing, the preferred embodiments of the present invention are described in detail.
The preparation of embodiment 1, silk gum-polyvinyl alcohol gel
Scheme 1：
The preparation method of porous silk gum/polyvinyl alcohol gel, comprises the following steps：
(1) extraction of sericin：Silk cocoon is cut into fractionlet, pressure cooker high temperature high pressure (120 DEG C, 0.1Mpa) is placed in Silk gum solution is prepared, by the silk gum solution filtered through gauze of preparation, removes comprising the insoluble substance including fibroin, obtains silk gum Solution, obtained silk gum solution is freeze-dried, silk gum powder is obtained；
(2) preparation of porous silk gum/polyvinyl alcohol gel：The silk that mass fraction is 2% is made in silk gum powder weight after molten Sol solution, the PVA solution for being then 2% with mass fraction is 4 by volume：1 mixing after add mould in, in room temperature (18~ 25 DEG C) place 10min after be put into multigelation 3-4 time in -20 DEG C of refrigerators, be then freeze-dried, obtained porous silk gum/gather Ethene alcogel.
In the same manner, difference is that multigelation is carried out under the conditions of -80 DEG C and 196 DEG C respectively.
Scheme 2：
The preparation method of porous silk gum/polyvinyl alcohol gel, comprises the following steps：
(1) extraction of sericin：Silk cocoon is cut into fractionlet, pressure cooker high temperature high pressure (120 DEG C, 0.1Mpa) is placed in Silk gum solution is prepared, by the silk gum solution filtered through gauze of preparation, removes comprising the insoluble substance including fibroin, obtains silk gum Solution, obtained silk gum solution is freeze-dried, silk gum powder is obtained；
(2) preparation of porous silk gum/polyvinyl alcohol gel：The silk that mass fraction is 1% is made in silk gum powder weight after molten Sol solution, the PVA solution for being then 1% with mass fraction is 4 by volume：2 mixing after add mould in, in room temperature (18~ 25 DEG C) place 10min after be put into multigelation in -80 DEG C of refrigerators, be then freeze-dried, obtained porous silk gum/polyethylene Alcogel.
In the same manner, difference is that the volume ratio of silk gum solution and PVA solution is respectively 4:1 and 4:4.
Scheme 3：
The preparation method of porous silk gum/polyvinyl alcohol gel, comprises the following steps：
(1) extraction of sericin：Silk cocoon is cut into fractionlet, pressure cooker high temperature high pressure (120 DEG C, 0.1Mpa) is placed in Silk gum solution is prepared, by the silk gum solution filtered through gauze of preparation, removes comprising the insoluble substance including fibroin, obtains silk gum Solution, obtained silk gum solution is freeze-dried, silk gum powder is obtained；
(2) preparation of porous silk gum/polyvinyl alcohol gel：The silk that mass fraction is 5% is made in silk gum powder weight after molten Sol solution, the PVA solution for being then 5% with mass fraction is 4 by volume：2 mixing after add mould in, in room temperature (18~ 25 DEG C) place 10min after be put into multigelation in -80 DEG C of refrigerators, be then freeze-dried, obtained porous silk gum/polyethylene Alcogel.
The hole situation of porous silk gum/polyvinyl alcohol gel made from different freezing conditions is detected, as a result as shown in Figure 1. It is different freezing bars that A, which observes B in the hole of the silk gum/polyvinyl alcohol gel prepared under different freezing conditions, Fig. 1 for SEM, in Fig. 1 The pore size statistics of the porous silk gum/polyvinyl alcohol gel prepared under part, as a result shows, obtained many under different freezing conditions The aperture of hole silk gum/polyvinyl alcohol gel is different, and temperature is lower, and aperture is smaller.C is molten for silk gum under -80 DEG C of freezing conditions in Fig. 1 The hole deviation rate for porous silk gum/polyvinyl alcohol gel that liquid is prepared with PVA solution different proportion, as a result shows, silk gum solution and PVA Solution has higher hole deviation rate under different proportion.
Fig. 2 is the sign of porous silk gum/polyvinyl alcohol gel；A is that FT-IR detects surface characteristics；B is mechanics properties testing As a result；C is that XRD detects crystalline structure；D is that TGA detects stability.As a result show, silk gum is well mixed with PVA, add after PVA The mechanical property of silk gum can be significantly improved, and does not influence the crystalline structure of silk gum, stability is improved.
Fig. 3 is porous silk gum/polyvinyl alcohol gel water imbibition test；A is porous silk gum/PVA contact angle results, shows many Hole silk gum/PVA quickly absorbs water, and B is porous silk gum/PVA swelling equilibrium times, and it is the main thing quickly absorbed water to show silk gum Matter；C is silk gum/PVA water absorption rates, shows largely absorb water in acid or alkali environment.
Fig. 4 is porous silk gum/polyvinyl alcohol gel fluoroscopic examination result.As a result show that silk gum forms mutually perforation with PVA solidifying Glue.
Embodiment 2, porous silk gum/polyvinyl alcohol gel carrying medicament
Scheme 1：
The method of porous silk gum-polyvinyl alcohol gel carrying medicament, comprises the following steps：
Porous silk gum/polyvinyl alcohol gel made from the scheme 1 of embodiment 1 be infiltrated on gentamicin concentration for 0.05mg/mL, Aspirin concentrations for 0.2mg/mL mixed solution in, obtain porous silk gum/polyvinyl alcohol gel of carrying medicament.
Scheme 2：
The method of porous silk gum-polyvinyl alcohol gel carrying medicament, comprises the following steps：
Porous silk gum/polyvinyl alcohol gel made from the scheme 1 of embodiment 1 be infiltrated on gentamicin concentration for 0.03mg/mL, Aspirin concentrations for 0.2mg/mL mixed solution in, obtain porous silk gum/polyvinyl alcohol gel of carrying medicament.
Scheme 3：
The method of porous silk gum-polyvinyl alcohol gel carrying medicament, comprises the following steps：
Porous silk gum/polyvinyl alcohol gel made from the scheme 1 of embodiment 1 be infiltrated on gentamicin concentration for 0.03mg/mL, Aspirin concentrations for 0.5mg/mL mixed solution in, obtain porous silk gum/polyvinyl alcohol gel of carrying medicament.
Fig. 5 is the testing result of porous silk gum/polyvinyl alcohol gel of carrying medicament；A is the standard curve of gentamicin, B is the standard curve of aspirin, and C is the release profiles of gentamicin, and D is the release profiles of aspirin.
Embodiment 3, porous silk gum/polyvinyl alcohol gel antibacterial test experience of carrying medicament
1. antibacterial growth curve experiment
The present invention by the bacterium of growth naturally and the porous silk gum/polyvinyl alcohol gel of addition, carrying medicament it is porous The growth curve of bacteria of silk gum/polyvinyl alcohol gel is contrasted, so that it is determined that silk gum/polyvinyl alcohol gel that medicine is loaded Antibacterial effect, specific method is：
(1) single bacterium colony of Escherichia coli and staphylococcus aureus is taken to be inoculated in the 100mL LB Liquid Cultures of sterilizing respectively It is that 220rpm, temperature are to cultivate 12 hours under the conditions of 37 DEG C in rotating speed in base (pH 7.4)；
(2) Escherichia coli and the μ L of staphylococcus aureus bacteria suspension 100 for taking step (1) activation respectively are added to 8mL LB In culture medium, every kind of bacterium solution prepares 5 groups, and one of which is blank group, and remaining 4 groups are separately added into porous silk gum/polyvinyl alcohol and coagulate Porous silk gum/polyvinyl alcohol gel (silk gum of glue, carrying medicament：Polyvinyl alcohol gel=4：1), the porous yarn of carrying medicament Glue/polyvinyl alcohol gel (silk gum：Polyvinyl alcohol gel=4：2), porous silk gum/polyvinyl alcohol gel (silk gum of carrying medicament： Polyvinyl alcohol gel=4：4) in rotating speed it is then, that 220rpm, temperature are culture under the conditions of 37 DEG C, and is taken in 0h, 12h, 24h Bacteria suspension 0.5mL, detects its absorbance value at 600nm, and Escherichia coli are drawn respectively and golden yellow according to the absorption value measured The staphylococcic growing state of color, as a result as shown in A in Fig. 6 and B.Analyze porous silk gum/polyvinyl alcohol gel, carrying medicament Influence of the porous silk gum/polyvinyl alcohol gel to bacterial growth, as a result shows, adds porous silk gum/polyethylene of drug loading After alcogel, the growth of Escherichia coli and staphylococcus aureus receives obvious suppression, shows load prepared by the present invention Porous silk gum/polyvinyl alcohol gel of medicine has excellent anti-microbial property.
2. Bactericidal test
For the antibacterial action for the porous silk gum/polyvinyl alcohol gel for further determining that carrying medicament, the side of inhibition zone is utilized Method tests fungistatic effect of the porous silk gum/polyvinyl alcohol gel to Escherichia coli and staphylococcus aureus of carrying medicament. Specific method is as follows：
(1) single bacterium colony of Escherichia coli and staphylococcus aureus is taken to be inoculated in the 100mL LB Liquid Cultures of sterilizing respectively It is that 220rpm, temperature are to cultivate 10 hours under the conditions of 37 DEG C in rotating speed in base (pH 7.4)；
(2) bacteria suspension for activating step (1), which dilutes, takes 200-300 μ L to add LB solid culture primary surfaces after 100 times, and Cultivated 1-2 hours in rotating speed is 220rpm shaking table, dilution is evenly distributed on agar media surface；
(3) a diameter of 0.3cm porous silk gum/polyvinyl alcohol gel, porous silk gum/polyvinyl alcohol of carrying medicament is taken to coagulate Glue (silk gum：Polyvinyl alcohol gel=4：1), porous silk gum/polyvinyl alcohol gel (silk gum of carrying medicament：Polyvinyl alcohol gel =4：2), porous silk gum/polyvinyl alcohol gel (silk gum of carrying medicament：Polyvinyl alcohol gel=4：4), it is laid in dilution point The uniform LB media surfaces of cloth；Then 12h is cultivated under the conditions of 37 DEG C, as a result as shown in C in Fig. 6 and D.As a result show, In the LB culture mediums for being placed with porous silk gum/polyvinyl alcohol gel, the culture medium and the golden yellow grape of inoculation of Escherichia coli are inoculated with Obvious inhibition zone is not formed on the culture medium of coccus, shows the growth of porous silk gum/polyvinyl alcohol gel to bacterium in itself There is no inhibitory action, and in the LB culture mediums of porous silk gum/polyvinyl alcohol gel containing carrying medicament, all observe and form Obvious inhibition zone, shows that silk gum/PVA film that medicine is loaded substantially can even be killed carefully to the growth inhibition effect of bacterium Bacterium.
The cytotoxicity experiment of embodiment 4, porous silk gum/polyvinyl alcohol gel of carrying medicament
In order to determine porous silk gum/polyvinyl alcohol gel biocompatibility of carrying medicament, cytotoxicity experiment has been carried out, Selected cell is MEC (NIH3T3) and human embryonic kidney cell (HEK293).Specific method is：
1st, NIH/3T3 and HEK293 cell culture is trained in the DMEM containing 10% (v/v) hyclone (FBS, Gibco) Support in base, containing 5%CO2, cultivate in 37 DEG C of incubators；
2nd, collect grow to NIH/3T3 the and HEK293 cells of 96% degree of converging tiled by 500/100uL/ holes it is thin to 48 holes In born of the same parents' culture plate, containing 5%CO2, 12h is cultivated in 37 DEG C of incubators；
3rd, porous silk gum/polyvinyl alcohol gel of 0.3cm porous silk gum/polyvinyl alcohol gel, carrying medicament is added Into 48 orifice plates, containing 5%CO2, continue in 37 DEG C of incubators to cultivate 12h, 24h and 36h；
4th, CCK-8 reagents are added according to the dosage in 20 μ L/ holes into 48 orifice plates, containing 5%CO2, 37 DEG C of CMC models 1.5h, determines the cell light absorption value in hole under 450nm wavelength.As a result as shown in A in Fig. 7 and B, porous silk gum/polyvinyl alcohol Gel cell growth does not influence, and porous silk gum/polyvinyl alcohol gel cell growth of carrying medicament does not also influence, table Bright no cytotoxicity.
5th, while carrying out life or death cell dyeing detection to cell.As a result as shown in C in Fig. 7 and D, porous silk gum/ Cell after PVA processing is compared with the cell after porous silk gum/polyvinyl alcohol gel processing of carrying medicament with control group The state of living cells, does not observe the presence of dead cell substantially, shows no cytotoxicity.
Embodiment 5, wound infection model experiment
In order to determine porous silk gum/polyvinyl alcohol gel practice of carrying medicament, wound infection model experiment is carried out, Selected MEC (NIH/3T3) and human embryonic kidney cell (HEK293) are used as infection cell.Specific method is：
1. by NIH/3T3 and HEK293 cell culture in 24 orifice plates containing 10% (v/v) hyclone (FBS, Gibco in DMEM culture mediums), 5%CO2, culture 24h treats that it grows up to cell monolayer in 37 DEG C of incubators；
2. suctioning out the culture medium in 24 orifice plates, and washed three times with PBS, add fresh nonreactive culture medium, individual layer now Cell simulates trauma skin.10 μ L staphylococcus aureuses are added into every hole cell, the bacterium simulated infection wound of addition Bacterium.Last porous silk gum/polyvinyl alcohol gel simulation wound dressing (Fig. 8, A) that carrying medicament is inserted on culture medium；
3. wound infection model is containing 5%CO2, to draw the detection of 10 μ L culture mediums coated plates thin by culture 12h in 37 DEG C of incubators The vigor of bacterium, CCK-8 reagents detect the activity of cell, while dead cell stain detects the survival condition of cell, as a result as in Fig. 8 Shown in B and C；As a result show add dressing experimental group Survival probability of bacteria is extremely low compared with the control and cell survival rate is high.
4. life or death cell dyeing testing result is as shown in D in Fig. 8 and F.As a result show, add the cell of the experimental group of dressing Substantially survive, seldom see dead cell, and experimental group does not observe living cells substantially.In coated plate testing result such as Fig. 8 E and Shown in G, as a result show in control group there is substantial amounts of bacterium, and do not observe bacterium in experimental group substantially then.Experimental result table Porous silk gum/polyvinyl alcohol gel of bright carrying medicament can protect cells from bacterium infect and bacterium can be killed and Cell growth does not influence.
Finally illustrate, preferred embodiment above is merely illustrative of the technical solution of the present invention and unrestricted, although logical Cross above preferred embodiment the present invention is described in detail, it is to be understood by those skilled in the art that can be Various changes are made to it in form and in details, without departing from claims of the present invention limited range.

Claims (10)

1. the preparation method of porous silk gum/polyvinyl alcohol gel, it is characterised in that comprise the following steps：By silk gum solution with gathering Added after glycohol solution mixing in mould, multigelation to aperture is formed at less than 0 DEG C and higher than 0 DEG C after standing, then It is freeze-dried, obtains porous silk gum/polyvinyl alcohol gel.
2. the preparation method of porous silk gum/polyvinyl alcohol gel according to claim 1, it is characterised in that：The silk gum is molten The mass fraction of liquid is 1~5%；The mass fraction of the poly-vinyl alcohol solution is 1~5%.
3. the preparation method of porous silk gum/polyvinyl alcohol gel according to claim 1, it is characterised in that：The silk gum is molten The volume ratio of liquid and poly-vinyl alcohol solution is 1：1~4：1.
4. the preparation method of porous silk gum/polyvinyl alcohol gel according to claim 1, it is characterised in that：It is described to freeze repeatedly Melt at -20 DEG C and 18~25 DEG C alternately；Or at -80 DEG C and 18~25 DEG C alternately；Or in -196 DEG C and 18 At~25 DEG C alternately.
5. the preparation method of porous silk gum/polyvinyl alcohol gel according to claim 1, it is characterised in that：The silk gum is molten Liquid is prepared by following methods：Silk cocoon is cut into fractionlet, handles, then with filtered through gauze, removes under the conditions of 120 DEG C, 0.1Mpa Insoluble substance is removed, is freeze-dried, silk gum powder, the molten obtained silk gum solution of weight is obtained.
6. porous silk gum/polyvinyl alcohol gel as made from any one of Claims 1 to 5 preparation method.
7. the porous silk gum/polyvinyl alcohol for preparing carrying medicament using porous silk gum/polyvinyl alcohol gel described in claim 6 coagulates Glue.
8. porous silk gum/polyvinyl alcohol gel of carrying medicament according to claim 7, it is characterised in that：The medicine is Gentamicin and aspirin.
9. the preparation method of porous silk gum/polyvinyl alcohol gel of carrying medicament described in claim 7 or 8, it is characterised in that：Will Porous silk gum/the polyvinyl alcohol gel is infiltrated in the solution containing medicine, obtains porous silk gum/polyvinyl alcohol of carrying medicament Gel.
10. application of the porous silk gum/polyvinyl alcohol gel of carrying medicament described in claim 7 or 8 in antiseptic dressing is prepared.
CN201710288164.2A 2017-04-27 2017-04-27 Preparation method of porous silk gum/polyvinyl alcohol gel and products thereof and application Pending CN107082907A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109054266A (en) * 2018-08-14 2018-12-21 河南工程学院 A kind of silk gum composite membrane and preparation method thereof
CN109395154A (en) * 2018-11-06 2019-03-01 东北林业大学 A kind of high porosity carries the preparation method of medicine wound dressing
CN114668883A (en) * 2022-04-21 2022-06-28 安徽大学 Preparation method of curcumin-loaded sodium alginate/polyvinyl alcohol/silk fibroin composite scaffold

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101028536A (en) * 2007-03-02 2007-09-05 上海交通大学 Production of polyvinyl alcohol/sericin blended gel thin film
CN105903057A (en) * 2016-04-14 2016-08-31 西南大学 Preparation method of nano-silver hybrid sericin porous gel antibacterial material, product and application thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101028536A (en) * 2007-03-02 2007-09-05 上海交通大学 Production of polyvinyl alcohol/sericin blended gel thin film
CN105903057A (en) * 2016-04-14 2016-08-31 西南大学 Preparation method of nano-silver hybrid sericin porous gel antibacterial material, product and application thereof

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109054266A (en) * 2018-08-14 2018-12-21 河南工程学院 A kind of silk gum composite membrane and preparation method thereof
CN109395154A (en) * 2018-11-06 2019-03-01 东北林业大学 A kind of high porosity carries the preparation method of medicine wound dressing
CN114668883A (en) * 2022-04-21 2022-06-28 安徽大学 Preparation method of curcumin-loaded sodium alginate/polyvinyl alcohol/silk fibroin composite scaffold
CN114668883B (en) * 2022-04-21 2023-02-28 安徽大学 Preparation method of curcumin-loaded sodium alginate/polyvinyl alcohol/silk fibroin composite scaffold

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Application publication date: 20170822