CN1887359A - Skin wound repairing agar/collagen dressing and its prepn and application - Google Patents
Skin wound repairing agar/collagen dressing and its prepn and application Download PDFInfo
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- CN1887359A CN1887359A CN 200610036652 CN200610036652A CN1887359A CN 1887359 A CN1887359 A CN 1887359A CN 200610036652 CN200610036652 CN 200610036652 CN 200610036652 A CN200610036652 A CN 200610036652A CN 1887359 A CN1887359 A CN 1887359A
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Abstract
The present invention provides one kind of skin wound repairing agar/collagen dressing and its preparation process and application. Agar and collagen or gelatin are mixed, and the mixture is freeze dried to obtain porous compound rack or compound film for promoting the repair of skin wound, and the rack may be also used in other soft tissue engineering. The porous compound rack and compound film of present invention has simple operation, low cost and wide use, and may be used in repairing burnt and ulcer skin as well as for cartilage, cornea, fat and soft tissue engineering.
Description
Technical field
The present invention relates to the biomedical material technology, be meant a kind of skin wound repairing agar/collagen dressing and its production and application especially.
Background technology
The dermatosis that large-area burns and ulcer etc. cause is common clinically disease, and skin reproduces or transplants is one of important measures of saving patient's life.Because for transplanting quantity not sufficient to solve the large tracts of land reparation, there are problems such as immunoreation in allosome or xenogenesis skin from the body micromicro, therefore all kinds of tissue engineering skins or the dressing of biomaterial class become the approach of dealing with problems.
The application of dressing for skin starts from B.C., and Plant fiber plants such as cotton fiber play the effect of good protection wound surface, promotion healing as the dressing of skin trauma to the wound of small size.Real both at home and abroad research started from the beginning of last century, and main path is that the xenogenesis skin substitutes, and has developed various biopolymer or the synthetic high polymer various dressing for skin for the basis afterwards, as chondroitin sulfate/collagen, and collagen/nylon, fibroin etc.The eighties in last century, the development speed of organization engineering skin is rapid, is the industrialization that " activity " skin of purpose has been realized limited scale to accelerate wound repair speed.The subject matter that restricts this industry development is selection cost height, and viable skin is not only with high costs, and repair mechanism is very not clear and definite.This patent uses inexpensive natural polymer-agar and derivant thereof, develops extensive wound reparation dressing for skin.
Utilize the design principle of organizational project, can obtain the film material and have three-dimensional porous interlayer structure, ensure cell proliferation, the needed nutrition supply of the normal healing of tissue, the disappearance and this film is degraded in organization healing, and the propagation and the differentiation of the monosaccharide pair cell of degraded have facilitation, make it really play the effect that guarantees the skin histology healing.Owing to significantly improve the surcharge of Sargassum polysaccharides-agar, the captivation that tangleweed is cultured further improves (agar extracts) from tangleweed, be expected to play a positive role in the restoration of the ecosystem of coastal waters simultaneously.Therefore because the prices of raw and semifnished materials of using are cheap, be expected to make product to be used widely as an important selection of skin trauma prosthesis, its economic benefit and social benefit are with highly significant.
Abroad in the document, except common biomaterial or acellular dermal matrix as the dressing for skin, also carry out the marketization of " activity " dressing for skin.Before more than 20 year, people such as the Bell of MIT observe form corium cell---fibroblast can penetrate in the collagen gel, and it can be changed into fibrous substrate alive.Collagen is the main constituent of extracellular matrix, and this biology " glue " can make the fibroblast location.According to this discovery, in May, 1998, the Apligraf with epidermis and corium bilayer skin structure that is developed by Organogenesis company becomes first by the implanting device that contains people's living cells of drugs approved by FDA, and this technology also is able to commercialization finally.Apligraf is mainly used in burn, diabetic ulcer and dermopathic surgical intervention.After U.S. Advanced Tissue Sciences company (La Jolla, California) isolates fibroblast from the neonate foreskin, be planted in two weeks of amplification on the polymer support.Cell in the division growth, is gone back protein components such as the synthetically grown factor, collagen in support, form functional artificial corium TransCyte.This product is ratified through FDA, is used for the treatment of 2～3 degree burns.Advanced TissueSciences company has developed monolayer corium Dermagraf subsequently again.Dermagraft is a living tissue, is applicable to the situation that need induce new skin growth, and the treatment as serious burn and diabetics skin ulcer also has better curative effect to chronic skin ulcer.Because Dermagraft contains living cells, need and store-70 ℃ of frozen back transportations.Need thaw before using, and be cut into certain shape.The cell of Dermagraft recovery back 50% can survive.Diabetics to 50 people group carries out clinical trial, and wherein 50% ulcer patient has recovered health, and matched group (general treatement) cure rate has only 8%.The patient who cures with Dermagraft does not see recurrence through 4 months to 1 year tracking.These products all are based on supports such as collagen/chondroitin sulfate, collagen/nylon, collagen/silicone rubber at present, and the market of product about annual 100 people of case, is in the hard state of dimension of taking a step 1,000,000 dollars of U.S.'s less thaies.
Therefore, used material does not all adopt agar and derivant thereof in the existing skin repair technology, and in view of the one-tenth gelling properties of agar, some patents only relate to agar as cooling, tackify dressing or as work such as cell encapsulations.
Summary of the invention
Select problem at the material that prior art exists, primary and foremost purpose of the present invention is to provide a kind of skin wound repairing agar/collagen dressing (being porous compound support frame or laminated film), this reparation dressing has three-dimensional porous interlayer structure, ensure the needed nutrition supply of the normal healing of skin histology, catabolite has the effect that promotes cell proliferation and express substrate simultaneously, and this porous compound support frame (or laminated film) disappearance of degrading in organization healing makes it really play the effect that promotes that skin trauma is repaired.
Another object of the present invention is to provide the preparation method of above-mentioned skin wound repairing agar/collagen dressing (porous compound support frame or laminated film).
A further object of the present invention is to provide the application of above-mentioned skin wound repairing agar/collagen dressing (porous compound support frame or laminated film).
The present invention is with collagen (collagen or the modifier that comprise I and types such as II) or gelatin (solubility) blend postlyophilization obtains porous compound support frame or convection drying becomes thin film by agar (comprising agarose and derivant thereof), to obtain a kind of skin trauma being repaired favorable support or thin film, provide support for other soft tissue engineering simultaneously.
Purpose of the present invention is achieved through the following technical solutions: the preparation method of a kind of skin wound repairing agar/collagen dressing (porous compound support frame or laminated film) comprises the steps:
(1) agar is dissolved in 85～95 ℃ of hot water, mass concentration is about 1%; With collagen gelatin swells in or the acetum of water-soluble or 0.04～0.06M in, mass concentration is 0.5～0.6%;
(2) above-mentioned two kinds of solution are mixed in proportion and water bath condition under stir 3～20 minutes after, 55～70 ℃ of described bath temperatures are poured in the mould after the vacuum or the centrifugal degassing, described agar and collagen or gelatin mass ratio be between 10: 1～1: 10 the variation;
(3) mould is placed 50～55 ℃ of dryings promptly obtained laminated film in 48～72 hours;
Perhaps mould is placed rapidly-18 ℃ freezing 2～4 hours, lyophilization then, cryogenic temperature-10～-70 ℃, freezing 1～3 hour, vacuum was bled 17～24 hours for following 0 ℃, promptly obtained porous compound support frame.
In order to realize the present invention better, can select lightly crosslinkedly according to instructions for use, porous compound support frame or laminated film mass concentration are 0.5～2% glutaraldehyde cross-linking 0.5～6 hour; 50～55 ℃ of dryings in distilled water washing back 48～72 hours promptly obtain porous compound support frame or laminated film.
The mixed proportion of described agar and collagen or gelatin and state (porous compound support frame or laminated film) are determined according to purposes, preferably: when porous compound support frame is used for skin trauma reparation dressing, can select about 3～4 millimeters of thickness for use, agar and collagen or gelatin mass ratio are 1: 1～1: 3, and be aided with lightly crosslinkedly, simultaneously such support can be used for soft tissue engineering scaffold; When laminated film is used for skin trauma reparation dressing, about 2～3 millimeters of thickness, agar and collagen or gelatin mass ratio are 1: 2～1: 5.
Described lyophilization condition optimization is :-35～-45 ℃ were descended freezing 1～3 hour, and vacuum was bled 17～24 hours for following 0 ℃.About 2～the 3mm of described THIN COMPOSITE thickness; The thick about 3～4mm of described porous compound support frame.
Use oxirane disinfection after resulting porous compound support frame or the laminated film cutting, get the finished product envelope and preserve; If need porous compound support frame or laminated film area big, can adopt glutaraldehyde stifling processing sterilization half an hour standby.
A kind of skin wound repairing agar/collagen dressing (porous compound support frame or laminated film) is exactly to be prepared from by above-mentioned preparation method.
The application of above-mentioned skin wound repairing agar/collagen dressing (porous compound support frame or laminated film) in skin trauma reparation dressing or soft tissue engineering scaffold material etc.
The present invention compared with prior art has following advantage and beneficial effect:
(1) porous compound support frame or the laminated film that obtain of the present invention can make the nutrition supply of healing district tissue unaffected, keeps the normal healing mechanism of tissue, and the propagation of catabolite pair cell and matrix group harness have facilitation, accelerates the generation of organizing.
(2) cost of the present invention is low, has improved the surcharge of agar and derivant thereof simultaneously, helps the plantation (agar extracts from tangleweed) of tangleweed, helps the rich restoration of the ecosystem of supporting in offshore sea waters.
(3) the present invention is easy to operate, and this porous compound support frame or laminated film intensity height are approaching with the skin tensile strength, pliability is good, can carry out sew application, can select the porous compound support frame or the laminated film of proper strength and degradation time according to the difference of instructions for use.
(4) agar/collagen porous compound support frame of the present invention or laminated film are of many uses, can be used as burn, the reparation of the big large tracts of land skin trauma of ulcer, and are used for soft tissue engineering scaffold materials such as cartilage, cornea, fat.
The specific embodiment
The present invention is described in further detail below in conjunction with embodiment, but embodiments of the present invention are not limited thereto.
Preparation of embodiment 1 porous compound support frame and dressing for skin
(1) agar is dissolved in 85～95 ℃ of hot water, and mass concentration percentage ratio is 1%, and collagen (or gelatin) is dissolved in the acetum (or swelling in water) of 0.04～0.06M, and mass concentration percentage ratio is 0.5%～0.6%.
(2) agar after step (1) processing and collagen (or gelatin) solution are pressed mass ratio mixing in 1: 2, stir after 10 minutes (70 ℃ of bath temperatures) under the water bath condition, (500～1000 rev/mins of the centrifugal degassings, 2～5 minutes), (making mixed liquor is 5～6mm) at the mould camber, and mould was put into refrigerator (18 ℃) freezing 2～4 hours to pour die for molding then into.
(3) mould is put into freezer dryer, descended freezing 1～3 hour, bled 17～24 hours at 0 ℃ then at-35～-45 ℃, be lyophilized into porous compound support frame, thick about 3～4mm, glutaraldehyde (mass concentration is 0.5%) crosslinked half an hour, oxirane disinfection is standby.
Preparation porous compound support frame diameter 1.5 centimetres of disk shapes, thick 3～4 millimeters, glutaraldehyde (mass concentration is 0.5%) crosslinked half an hour, oxirane disinfection is standby.With rabbit skin wound surface is model, and it is damaged to make 1.5 cm diameters, and sutured is implanted 2～3 all backs skin repair and finished scaffold degradation.
Preparation of embodiment 2 composite membranes and dressing for skin
(1) agar is dissolved in 85～95 ℃ of hot water, and mass concentration percentage ratio is 1%, and gelatin (or collagen) is dissolved in the acetum (or swelling in water) of 0.04～0.06M, and mass concentration percentage ratio is 0.5%～0.6%.
(2) agar after step (1) processing and gelatin (or collagen) solution are pressed mass ratio mixing in 1: 4, stir after 3 minutes (70 ℃ of bath temperatures) under the water bath condition, (500～1000 rev/mins of the centrifugal degassings, 2～5 minutes), (making mixed liquor is 3～4mm) at the mould camber to pour die for molding then into.
(3) put into blast drier, drying is 48～72 hours under 50～55 ℃, obtains composite membrane, the about 2mm of thickness.Oxirane disinfection is standby.
Preparation composite film material diameter 1.5 centimetres of disk shapes, thick 2～3 millimeters, glutaraldehyde (mass concentration is 0.5%) crosslinked half an hour, oxirane disinfection is standby.With rabbit skin wound surface is model, and it is damaged to make 1.5 cm diameters, and sutured is implanted 3 all backs skin repair and finished scaffold degradation.
Embodiment 3 subcutaneus adipose tissue engineering racks
(1) agar is dissolved in 85～95 ℃ of hot water, and mass concentration percentage ratio is 1%, and collagen (or gelatin) swells in the water (or be dissolved in 0.04～0.06M acetum), and mass concentration percentage ratio is 0.5%～0.6%.
(2) agar after step (1) processing and collagen (or gelatin) solution are pressed mass ratio mixing in 2: 1, stir after 20 minutes (60 ℃ of bath temperatures) under the water bath condition, (500～1000 rev/mins of the centrifugal degassings, 2～5 minutes), (making mixed liquor is 5～6mm) at the mould camber, puts into refrigerator (18 ℃) freezing 2～4 hours to pour die for molding then into.
(3) put into freezer dryer, descended freezing 2 hours, bled 20 hours at 0 ℃ then, be lyophilized into porous compound support frame, thick about 3～4mm at-40 ℃.
(4) be to fumigate on 2% glutaraldehyde water solution to carry out crosslinking Treatment half an hour with porous compound support frame in mass concentration.
0.8 centimetre of the porous compound support frame diameter of preparation, thick 3～4 millimeters, oxirane disinfection is standby.The subcutaneous adult fat stem cell 8,000,000 that increases is planted in support, air-dry 3 hour cells add conventional culture fluid (DMEM+10%FBS+1% antibiotics) (replenishing 10ng/ml bFGF) after attaching, fat-like soft tissue under the 3 week back cutifications, scaffold degradation.
Embodiment 4 skin tissue engineering scaffolds
(1) agar is dissolved in 85～95 ℃ of hot water, and mass concentration percentage ratio is 1%, and collagen (or gelatin) swells in the water (or be dissolved in 0.04～0.06M acetum), and mass concentration percentage ratio is 0.5%～0.6%.
(2) agar after step (1) processing and collagen (or gelatin) solution are pressed mass ratio mixing in 1: 10, stir after 20 minutes (60 ℃ of bath temperatures) under the water bath condition, (500～1000 rev/mins of the vacuum (vacuum 100Pa is following) or the centrifugal degassings, 2～5 minutes), (making mixed liquor is 5～6mm) at the mould camber, puts into refrigerator (18 ℃) freezing 2～4 hours to pour die for molding then into.
(3) put into freezer dryer, descended freezing 2 hours, bled 17 hours at 0 ℃ then, be lyophilized into porous compound support frame, thick about 3～4mm at-10 ℃.
(4) be to fumigate on 2% glutaraldehyde water solution to carry out crosslinking Treatment half an hour with porous compound support frame in mass concentration.
0.8 centimetre of the porous compound support frame diameter of preparation, thick 3～4 millimeters, oxirane disinfection is standby.Amplification epidermis fibroblast 8,000,000 is planted in support, air-dry 3 hour cells add conventional culture fluid (DMEM+10%FBS+1% antibiotics) (replenishing 10ng/ml bFGF) after attaching, 2 week the back become epiderm-like soft tissues, scaffold degradation.
Embodiment 5 anti-skin infection dressing
(1) agar is dissolved in 85～95 ℃ of hot water, and mass concentration percentage ratio is 1%, and collagen (or gelatin) swells in the water (or be dissolved in 0.04～0.06M acetum), and mass concentration percentage ratio is 0.5%～0.6%.
(2) agar after step (1) processing and collagen (or gelatin) solution are pressed mass ratio mixing in 10: 1, stir after 20 minutes (55 ℃ of bath temperatures) under the water bath condition, (500～1000 rev/mins of the centrifugal degassings, 2～5 minutes), (making mixed liquor is 3～4mm) at the mould camber, puts into refrigerator (18 ℃) freezing 2～4 hours to pour die for molding then into.
(3) put into freezer dryer, descended freezing 2 hours, bled 24 hours at 0 ℃ then, be lyophilized into porous compound support frame, thick about 2～3mm at-70 ℃.
(4) be to fumigate on 1% glutaraldehyde water solution to carry out crosslinking Treatment half an hour with porous compound support frame in mass concentration.Oxirane disinfection is standby.
The material of above-mentioned preparation is sticked in drug of topical application skin burn place, take material skin after one or two week off and recover normal, this material rises every the bacterium effect.
Claims (8)
1, a kind of preparation method of skin wound repairing agar/collagen dressing comprises the steps:
(1) agar is dissolved in 85～95 ℃ of hot water, mass concentration is 1%; Collagen or gelatin swell in or the acetum of water-soluble or 0.04～0.06M in, mass concentration is 0.5～0.6%;
(2) above-mentioned two kinds of solution are mixed in proportion and water bath condition under stir 3～20 minutes after, described bath temperature 55～70C pours in the mould after the vacuum or the centrifugal degassing, described agar and collagen or gelatin mass ratio are 10: 1～1: 10;
(3) mould is placed 50～55 ℃ of dryings promptly obtained laminated film in 48～72 hours;
Perhaps mould is placed-18 ℃ freezing 2～4 hours, lyophilization then, cryogenic temperature-10～-70 ℃, vacuum was bled 17～24 hours for following 0 ℃, promptly obtained porous compound support frame.
2, the preparation method of a kind of skin wound repairing agar/collagen dressing according to claim 1 is characterized in that, described porous compound support frame or laminated film mass concentration are 0.5～2% glutaraldehyde cross-linking 0.5～6 hour; 50～55 ℃ of dryings in distilled water washing back 48～72 hours obtain porous compound support frame or laminated film.
3, the preparation method of a kind of skin wound repairing agar/collagen dressing according to claim 1 is characterized in that, during the preparation porous compound support frame, agar and collagen or gelatin mass ratio are 1: 1～1: 3; During the preparation laminated film, agar and collagen or gelatin mass ratio are 1: 2～1: 5.
4, the preparation method of a kind of skin wound repairing agar/collagen dressing according to claim 1 is characterized in that, described lyophilization condition is :-35～-45 ℃ were descended freezing 1～3 hour, and vacuum was bled 17～24 hours for following 0 ℃.
5, the preparation method of a kind of skin wound repairing agar/collagen dressing according to claim 1 is characterized in that, described THIN COMPOSITE thickness 2～3mm; Thick 3～the 4mm of described porous compound support frame.
6, the preparation method of a kind of skin wound repairing agar/collagen dressing according to claim 1 is characterized in that, uses oxirane disinfection after porous compound support frame or the laminated film cutting, or standby with glutaraldehyde stifling processing sterilization half an hour.
7, a kind of skin wound repairing agar/collagen dressing is prepared from by the described preparation method of claim 1.
8, the application of the described skin wound repairing agar/collagen dressing of claim 7 in skin trauma reparation dressing or soft tissue engineering scaffold material.
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| CNB2006100366526A CN100402097C (en) | 2006-07-24 | 2006-07-24 | Skin wound repairing agar/collagen dressing and its prepn and application |
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| CNB2006100366526A CN100402097C (en) | 2006-07-24 | 2006-07-24 | Skin wound repairing agar/collagen dressing and its prepn and application |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103386145A (en) * | 2013-07-31 | 2013-11-13 | 暨南大学 | Wound healing dressing containing carrageenan, and preparation method and application of wound healing dressing |
| CN103948957A (en) * | 2014-05-14 | 2014-07-30 | 广州创尔生物技术股份有限公司 | Hydrophilic and hydrophobic agarose collagen dressing as well as preparation method and application thereof |
| CN105233337A (en) * | 2015-09-30 | 2016-01-13 | 苏州蔻美新材料有限公司 | Cartilage tissue repair membrane and preparation method thereof |
| CN108114306A (en) * | 2018-02-07 | 2018-06-05 | 苏州元禾医疗器械有限公司 | A kind of preparation method of wound dressing |
| CN110218342A (en) * | 2019-05-13 | 2019-09-10 | 暨南大学 | Agarose-gelatin graft temperature-sensitive hydrogel and the preparation method and application thereof |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN2304399Y (en) * | 1997-01-16 | 1999-01-20 | 重庆西南医院 | Slow degradated collagen membrane net for medical uses |
| CN1234429C (en) * | 2002-05-09 | 2006-01-04 | 中国人民解放军军事医学科学院野战输血研究所 | Artificial skin, preparing method and application thereof |
| JP4189487B2 (en) * | 2003-10-22 | 2008-12-03 | 独立行政法人産業技術総合研究所 | Composition for preparing biological models of skin, mucous membranes, etc. |
| US9526746B2 (en) * | 2004-02-13 | 2016-12-27 | Smith & Nephew, Inc. | Wound healing composition |
| CN100339135C (en) * | 2005-05-13 | 2007-09-26 | 褚省吾 | Antibiotic aerogel dressing of hollow acetate fiber and its prepn |
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- 2006-07-24 CN CNB2006100366526A patent/CN100402097C/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103386145A (en) * | 2013-07-31 | 2013-11-13 | 暨南大学 | Wound healing dressing containing carrageenan, and preparation method and application of wound healing dressing |
| CN103386145B (en) * | 2013-07-31 | 2015-03-04 | 暨南大学 | Wound healing dressing containing carrageenan, and preparation method and application of wound healing dressing |
| CN103948957A (en) * | 2014-05-14 | 2014-07-30 | 广州创尔生物技术股份有限公司 | Hydrophilic and hydrophobic agarose collagen dressing as well as preparation method and application thereof |
| CN103948957B (en) * | 2014-05-14 | 2015-12-09 | 广州创尔生物技术股份有限公司 | A kind of hydrophilic and hydrophobic agarose collagen dressing and its preparation method and application |
| CN105233337A (en) * | 2015-09-30 | 2016-01-13 | 苏州蔻美新材料有限公司 | Cartilage tissue repair membrane and preparation method thereof |
| CN108114306A (en) * | 2018-02-07 | 2018-06-05 | 苏州元禾医疗器械有限公司 | A kind of preparation method of wound dressing |
| CN110218342A (en) * | 2019-05-13 | 2019-09-10 | 暨南大学 | Agarose-gelatin graft temperature-sensitive hydrogel and the preparation method and application thereof |
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| CN100402097C (en) | 2008-07-16 |
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