CN1285675C - Composite collagen-base rack material and its pepn and use - Google Patents

Composite collagen-base rack material and its pepn and use Download PDF

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Publication number
CN1285675C
CN1285675C CN 03139063 CN03139063A CN1285675C CN 1285675 C CN1285675 C CN 1285675C CN 03139063 CN03139063 CN 03139063 CN 03139063 A CN03139063 A CN 03139063A CN 1285675 C CN1285675 C CN 1285675C
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collagen
base compound
support frame
skin
water
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CN 03139063
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CN1526764A (en
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曹成波
吴克安
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Shandong University
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Shandong University
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Abstract

The present invention relates to collagen base compound support material, a preparing method thereof and an application thereof. The material comprises collagen and water soluble polysaccharide, wherein the water soluble polysaccharide is at least one kind of chitosan, salt of seaweed polysaccharide, proteoglycan, cellulose, carrageenin, acacia and starch, and the collagen has the three-dimensional net structure of natural fiber. The method for preparing the support material comprises: mammal skin is used as raw material, a splitting machine is used for removing subcutaneous tissues, and then, the mammal skin is soaked in water, degreased, soaked in lime, delimed, split, softened, compounded, irradiated by cobalt 60 and sterilized. Functional material can be added into the collagen base compound support material to prepare biological dressings, manual skin, manual genuine skin and manual compound skin, and the collagen base compound support material can also be matched with ointment. The support material has the advantages of favorable permeability, favorable water and vapor permeability and favorable absorptive quality.

Description

A kind of collagen base compound support frame material and its production and use
Technical field
The present invention relates to chemical, medical science, materialogy, bioengineering field, particularly a kind of collagen base compound support frame material and its production and use.
Background technology
Skin tissue engineering can be divided into synthetic timbering material, biologic bracket material and this three major types of both compound support frame materials with timbering material.Synthetic materials tensile strength height, degradation rate and microstructure are controlled in building-up process easily, but its histocompatibility, cell affinity, water-absorbent, surface of a wound tackiness equal difference need be improved its performance with suitable chemistry and biological method.Poor as urethane timbering material water-absorbent, make the surface of a wound easily produce hydrops, bacterium easily breeds, and does not possess the growth of the cell of adjusting, promotes the function of wound healing.The biomaterial good biocompatibility is easy to cell attachment and growth, but degradation rate is fast, microstructure is wayward etc.Though can well promote the surface of a wound to recover as the collagen gel timbering material, poor mechanical property, very crisp when dry, easily broken behind the absorption wound exudate, and under the effect of collagenase, be easy to degraded, can not well play a protective role.The copolymer composite collagen gel of glycollide and rac-Lactide is as the artificial skin timbering material, though improved tensile strength, the small molecules after the polymer degradation forms the tart environment in the part, be unfavorable for the growth of cell.
The patent No. is US 5282, the double-layered artificial skin that 859 United States Patent (USP) is introduced, epidermal area is highly purified micropore collagen gel film, skin corium has fibroblastic porous collagen sponge for cultivating, glutaraldehyde cross-linking improves its tensile strength, but its tensile strength does not reach requirements for clinical application, and degradation rate is too fast, the glutaraldehyde pair cell is toxic.The patent No. is CN 1387,923 Chinese patent discloses a kind of heterogeneous cell-free hypodermal framework and preparation method thereof, this carriage support is handled the sucking pig skin with trypsinase and glutaraldehyde and is got, its tensile strength height, degradation rate are controlled, histocompatibility is good, but poor permeability, the surface of a wound easily produce hydrops, pneumatosis, and vascularization is slow.The patent No. is CN 1343,523 Chinese patent discloses a kind of micropore allosome or xenogenesis acellular dermis surrogate that can be used as wound repair, adopt the method for machinery or laser on acellular dermis, to stamp intensive perfoliate hole regularly, though this can solve the shortcoming of the poor permeability of acellular dermis surrogate, it is not thorough that but the non-fibre composition in the dermal tissue is removed, and its surface bioactive and absorptivity are poor.
Summary of the invention
The object of the invention is to overcome above-mentioned the deficiencies in the prior art, and a kind of natural fiber three-dimensional net structure collagen base compound support frame material with good penetration, absorptivity and surface bioactive and its production and use is provided.
The present invention realizes in the following manner:
A kind of collagen base compound support frame material, comprise collagen, it is characterized in that it also comprises water-soluble polysaccharide, water-soluble polysaccharide is at least a in the salt, protein-polysaccharide, Mierocrystalline cellulose, carrageenin, gum arabic, starch of chitin, Sargassum polysaccharides, and collagen has the natural fiber three-dimensional net structure.
Above-mentioned collagen base compound support frame material, the weight ratio that it is characterized in that water-soluble polysaccharide and collagen is 0.01-0.2: 1.
The preparation method of above-mentioned collagen base compound support frame material is: selecting skin of mammal is raw material, remove subcutis with splitting machine, immersion then, degreasing, liming, deliming, cut open layer, it is characterized in that it comprises that also (1) is softening: liquor ratio 1.0~1.5,35~40 ℃ of temperature, interfibrillar substance is fully removed in the pancreatin of adding raw material tare weight amount 0.1~0.3% and 0.1~0.3% neutral protease effect 30~240 minutes; (2) compound: liquor ratio 0.5~0.8,35 ℃ of temperature add the water-soluble polysaccharide of raw material tare weight amount 1~20%, compound 30~40 minutes; (3) sterilization: cobalt 60 radiation sterilizations.
The purposes of above-mentioned collagen base compound support frame material, it is characterized in that in collagen base compound support frame material adding functional material and can be used to prepare biological dressing, artificial epidermis, artificial dermis or artificial Composite Skin, wherein said functional material is that antiseptic-germicide, anti-mycotic agent, Wound-healing agent, somatomedin, skin basement membrane form at least a in promotor, the glycerine;
The purposes of above-mentioned collagen base compound support frame material, it is characterized in that the supporting use of collagen base compound support frame material and ointment, ointment is applied to the one side that contacts with the surface of a wound, and the ointment composition comprises at least two kinds in polypeptide, glycerine, antiseptic-germicide, anti-mycotic agent, hemostatic agent, oxygen free radical scavenger, the somatomedin.
Natural fiber three-dimensional net structure collagen has stronger tensile strength, for cell provides the growing space that is fit to, helps the self-sow and the arrangement of cell.Chitin has mechanical property preferably, is prepared into the microphase-separated vesicular structure easily, helps the adhesion of cell, can suppress bacterium and mould, promotion epithelial cell growth, promote wound healing.Protanal TXF 200 has extremely strong absorptivity, and can effectively control sepage and hemostasis.Hyaluronic acid is a kind of high-molecular weight straight chain protein-polysaccharide, and it impels wound healing by regulating the interaction of cell and matrix, suppresses the formation that the collagen over-deposit causes scar.It can also provide good outside atmosphere and nutritive substance for collagen protein and spandex fiber synthetic, stop the generation of some enzymes in the cell, the formation, particularly low-molecular-weight hyaluronic acid that reduces free radical have anti-inflammatory, antibacterial, promote effects such as cell growth and differentiation.
The present invention compared with prior art has the following advantages:
1. compound collagen base timbering material has favorable tissue consistency, complete degradation property and cell affinity.
2. compound collagen base timbering material has good penetration, permeability to water vapour and absorptivity.
3. compound collagen base timbering material still has tensile strength preferably under the situation that absorbs a large amount of wound fluids.
4. compound collagen base timbering material has breeding and the arrangement of regulating cell, promotes the vascularization of newborn skin, promotes the reparation and the regeneration of damaged skin.
5. compound collagen base timbering material can suppress scar proliferation, makes the outward appearance that has original skin behind the wound healing.
6. compound collagen base timbering material can be prepared into biological dressing, artificial epidermis, artificial dermis and artificial Composite Skin according to different requirements.
Embodiment:
Provide two most preferred embodiments of the present invention below:
1 example of embodiment provides a kind of composite collagen base timbering material that contains collagen, chitin, Sargassum polysaccharides, three kinds of material weight ratios are 100: 6: 3, choose salted pigskin and be raw material and according to size packets, with splitting machine the raw material skin is cutd open then and get the upper strata, remove subcutis, weigh as reagent dosage.
1. immersion: liquor ratio 3.0,40 ℃ of temperature, grease-removing agent 1% soaked 90 minutes, control water;
2. degreasing: liquor ratio 1.5,37~38 ℃ of temperature, soda ash 2% acts on 90 minutes, washing;
3. secondary degreasing: liquor ratio 2.5,37~38 ℃ of temperature, permeate agent: 0.3%, soda ash 1.5% changeed 120 minutes, control water, dialled hair;
4. buttocks is coated with enzyme: neutral protease (50,000 unit) 0.2%, and ammonium sulfate 0.3%, 0.8%, 30 ℃ of water mixing of wheat bran, denseness is advisable with stream not; 48~50 hours, flowing water washed down then;
5. liming: liquor ratio 0.8,18~20 ℃ of temperature, Sodium Sulphide (50%) 2.1%, lime powder (50%) 1.5%, auxiliary liming agent 1% acts on 120 minutes, and lime powder 2% changeed 30 minutes; Add 21~22 ℃ of water to total liquor ratio 2.5~3.0, rotated total time 15h 10 minutes in 50 minutes at interval;
6. washing: normal temperature flowing water was washed 40 minutes, did not send out cunning to skin;
7. cut open in the meat, and weigh, wash;
8. deliming: liquor ratio 0.6, temperature 25~26, ammonium sulfate 2%, ammonium chloride 1%, grease-removing agent 1%, deliming 120 minutes, pH value about 8.4;
9. washing: the normal temperature current were washed 30 minutes, and 50 ℃ of pre-temperature of water are controlled water purification to 35~36 ℃;
10. soak nitre: Sodium sulfate anhydrous.min(99) 8%, grease-removing agent 0.5% acts on 3 hours, soaks into;
11. water squeezing, cut open the layer, deburring;
12. weigh, be washed to clearly;
13. it is softening: liquor ratio 2.0,38 ℃ of temperature, pancreatin (2.5 times) 0.3%, neutral protease 0.1~0.3%, ammonium sulfate 0.3%, grease-removing agent 1%, softening 30~240 minutes;
14. it is compound: liquor ratio 0.5~1.0, normal temperature, polysaccharide 10%, compound 20~60 minutes, control water;
15. sterilization: cobalt 60 irradiation sterilizations;
One of purposes of collagen base compound support frame material by method for preparing is the preparation biological dressing, and application method is to add 0.5% antiseptic-germicide, 2% glycerine therein, then ointment of the present invention is applied to the one side that contacts with the surface of a wound and gets final product.
Embodiment 2 present embodiments provide a kind of collagen, chitin, hyaluronic composite collagen base timbering material of containing, and the weight ratio of three kinds of materials is 100: 5: 4, adopt the following steps preparation:
Press example 1 operation;
Compound: liquor ratio 0.8, normal temperature, chitin 5%, hyaluronic acid 4% rotated 30 minutes, control water;
Drying and sterilization are with example 1.
One of purposes of collagen base compound support frame material by method for preparing is the artificial epidermis of preparation, and its method is for adding 0.3% antiseptic-germicide, 0.2% anti-mycotic agent, growth factors 5 μ g/m therein 2, skin basement membrane forms promotor 10 μ g/m 2, 2% glycerine, ointment of the present invention is applied to the one side that contacts with the surface of a wound and is transplanted to the surface of a wound.Then people's foreskin is made single cell suspension with Dispase II-protease digestion, getting a part cultivates at serum-free epidermic cell substratum, carry out formerly being commissioned to train fosterly with serum-free epidermic cell substratum, adopt the 2nd acute pyogenic infection of finger tip to count the cell in vegetative period, become single cell suspension with tryptic digestion.And the single cell suspension for preparing is inoculated in is transplanted to behind the surface of a wound 1-2 days artificial skin surface, density is not less than 1 * 10 5/ cm 2

Claims (5)

1. collagen base compound support frame material, comprise collagen, it is characterized in that it also comprises water-soluble polysaccharide, water-soluble polysaccharide is at least a in the salt, protein-polysaccharide, Mierocrystalline cellulose, carrageenin, gum arabic, starch of chitin, Sargassum polysaccharides, and collagen has the natural fiber three-dimensional net structure.
2. collagen base compound support frame material according to claim 1, the weight ratio that it is characterized in that water-soluble polysaccharide and collagen is 0.01-0.2: 1.
3. the preparation method according to claim 1,2 described collagen base compound support frame materials is: selecting skin of mammal is raw material, remove subcutis with splitting machine, immersion then, degreasing, liming, deliming, cut open layer, it is characterized in that it comprises that also (1) is softening: liquor ratio 1.0~1.5,35~40 ℃ of temperature, interfibrillar substance is fully removed in the pancreatin of adding raw material tare weight amount 0.1~0.3% and 0.1~0.3% neutral protease effect 30~240 minutes; (2) compound: liquor ratio 0.5~0.8,35 ℃ of temperature add the water-soluble polysaccharide of raw material tare weight amount 1~20%, compound 30~40 minutes; (3) sterilization: cobalt 60 radiation sterilizations.
4. according to the purposes of claim 1,2 described collagen base compound support frame materials, it is characterized in that in collagen base compound support frame material adding functional material and can be used to prepare biological dressing, artificial epidermis, artificial dermis or artificial Composite Skin, wherein said functional material is that antiseptic-germicide, anti-mycotic agent, Wound-healing agent, somatomedin, skin basement membrane form at least a in promotor, the glycerine.
5. according to the purposes of claim 1,2 described collagen base compound support frame materials, it is characterized in that the supporting use of collagen base compound support frame material and ointment, ointment is applied to the one side that contacts with the surface of a wound, and the ointment composition comprises at least two kinds in polypeptide, glycerine, antiseptic-germicide, anti-mycotic agent, hemostatic agent, oxygen free radical scavenger, the somatomedin.
CN 03139063 2003-09-24 2003-09-24 Composite collagen-base rack material and its pepn and use Expired - Fee Related CN1285675C (en)

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Families Citing this family (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100464791C (en) * 2005-07-11 2009-03-04 山东大学 Method for preparing biological filler for injectable soft tissue
CN101864175B (en) * 2009-04-16 2012-05-23 西南科技大学 Thermoplastic animal protein/starch blending material and preparation method thereof
CN102108138B (en) * 2009-12-28 2012-07-25 沈阳鑫盛生物科技有限公司 Method for preparing chitosan collagen gel
CN101856516B (en) * 2010-05-25 2011-11-30 中国人民解放军总医院第一附属医院 Preparation of collagen-chitosan-laser micropore dermal matrix composite membranes
CN102772822A (en) * 2011-05-10 2012-11-14 中国人民解放军军事医学科学院卫生装备研究所 Application of collagen matrix as tissue engineering scaffold
CN102772819B (en) * 2012-05-09 2014-07-23 苏州博创同康生物工程有限公司 Skin wound biological induced active dressing, preparation method and application thereof
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
CN105268022B (en) * 2014-08-01 2018-04-20 山东大学 A kind of preparation method of xenogenesis cell-less corium ground substance
CN105288702B (en) * 2014-08-01 2018-11-02 山东大学 A kind of acellular polysaccharide dermal matrix material and its preparation method and application
CN105412989B (en) * 2015-12-07 2018-11-02 杭州华迈医疗器械有限公司 Acellular biological tissue matrix material-based composite material for bone repair and preparation method thereof
CN106890362B (en) * 2017-03-03 2020-07-07 山东大学 Pericardial collagen composite material and preparation method and application thereof
CN111393521A (en) * 2020-03-20 2020-07-10 张培华 Extraction method of jellyfish collagen

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