CN108014370A - A kind of promotion, which is press-offed, injures the collagen material that avulsion skin survives - Google Patents
A kind of promotion, which is press-offed, injures the collagen material that avulsion skin survives Download PDFInfo
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- CN108014370A CN108014370A CN201711313526.5A CN201711313526A CN108014370A CN 108014370 A CN108014370 A CN 108014370A CN 201711313526 A CN201711313526 A CN 201711313526A CN 108014370 A CN108014370 A CN 108014370A
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- collagen
- avulsion
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- protein sponge
- skin
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- 102000008186 Collagen Human genes 0.000 title claims abstract description 65
- 108010035532 Collagen Proteins 0.000 title claims abstract description 65
- 229920001436 collagen Polymers 0.000 title claims abstract description 38
- 239000000463 material Substances 0.000 title claims abstract description 20
- 235000015097 nutrients Nutrition 0.000 claims abstract description 16
- 238000004108 freeze drying Methods 0.000 claims abstract description 7
- 238000000746 purification Methods 0.000 claims abstract description 7
- 238000000034 method Methods 0.000 claims abstract description 5
- 238000000605 extraction Methods 0.000 claims abstract description 4
- 239000002994 raw material Substances 0.000 claims abstract description 4
- 239000000243 solution Substances 0.000 claims description 31
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 18
- 239000007788 liquid Substances 0.000 claims description 12
- 238000005119 centrifugation Methods 0.000 claims description 10
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 9
- 239000011780 sodium chloride Substances 0.000 claims description 9
- 239000003431 cross linking reagent Substances 0.000 claims description 8
- 210000001519 tissue Anatomy 0.000 claims description 8
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 6
- 238000000151 deposition Methods 0.000 claims description 6
- 230000008021 deposition Effects 0.000 claims description 6
- 238000004132 cross linking Methods 0.000 claims description 5
- 238000001556 precipitation Methods 0.000 claims description 5
- 238000012545 processing Methods 0.000 claims description 4
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 3
- 102000003971 Fibroblast Growth Factor 1 Human genes 0.000 claims description 3
- 108090000386 Fibroblast Growth Factor 1 Proteins 0.000 claims description 3
- 102000057297 Pepsin A Human genes 0.000 claims description 3
- 108090000284 Pepsin A Proteins 0.000 claims description 3
- 229960000583 acetic acid Drugs 0.000 claims description 3
- 230000002378 acidificating effect Effects 0.000 claims description 3
- 210000000577 adipose tissue Anatomy 0.000 claims description 3
- 239000007853 buffer solution Substances 0.000 claims description 3
- 239000008367 deionised water Substances 0.000 claims description 3
- 229910021641 deionized water Inorganic materials 0.000 claims description 3
- JMQGGPRJQOQKRT-UHFFFAOYSA-N diphenyl hydrogen phosphate;azide Chemical compound [N-]=[N+]=[N-].C=1C=CC=CC=1OP(=O)(O)OC1=CC=CC=C1 JMQGGPRJQOQKRT-UHFFFAOYSA-N 0.000 claims description 3
- 239000012362 glacial acetic acid Substances 0.000 claims description 3
- 239000003102 growth factor Substances 0.000 claims description 3
- 238000007654 immersion Methods 0.000 claims description 3
- 239000012535 impurity Substances 0.000 claims description 3
- 150000002632 lipids Chemical class 0.000 claims description 3
- 238000003760 magnetic stirring Methods 0.000 claims description 3
- 238000005360 mashing Methods 0.000 claims description 3
- 210000003205 muscle Anatomy 0.000 claims description 3
- 229940111202 pepsin Drugs 0.000 claims description 3
- 238000002203 pretreatment Methods 0.000 claims description 3
- 230000005855 radiation Effects 0.000 claims description 3
- 210000002966 serum Anatomy 0.000 claims description 3
- 210000004974 shell Anatomy 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- 239000008399 tap water Substances 0.000 claims description 3
- 235000020679 tap water Nutrition 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 244000144992 flock Species 0.000 claims 1
- 239000002244 precipitate Substances 0.000 claims 1
- 238000005406 washing Methods 0.000 claims 1
- 230000006378 damage Effects 0.000 abstract description 18
- 210000002435 tendon Anatomy 0.000 abstract description 16
- 210000000988 bone and bone Anatomy 0.000 abstract description 13
- 231100000429 cutaneous necrosis Toxicity 0.000 abstract description 7
- 238000001356 surgical procedure Methods 0.000 abstract description 6
- 238000011084 recovery Methods 0.000 abstract description 3
- 206010058046 Post procedural complication Diseases 0.000 abstract 1
- 208000035965 Postoperative Complications Diseases 0.000 abstract 1
- 208000027418 Wounds and injury Diseases 0.000 description 24
- 210000004369 blood Anatomy 0.000 description 16
- 239000008280 blood Substances 0.000 description 16
- 208000014674 injury Diseases 0.000 description 14
- 206010063560 Excessive granulation tissue Diseases 0.000 description 4
- 210000002950 fibroblast Anatomy 0.000 description 4
- 210000001126 granulation tissue Anatomy 0.000 description 4
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 206010033675 panniculitis Diseases 0.000 description 2
- 230000001376 precipitating effect Effects 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 108010048734 sclerotin Proteins 0.000 description 2
- 208000023929 Degloving injury Diseases 0.000 description 1
- 206010028851 Necrosis Diseases 0.000 description 1
- 206010072170 Skin wound Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000001804 debridement Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 239000012567 medical material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
Classifications
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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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/60—Materials for use in artificial skin
-
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/24—Collagen
-
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
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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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
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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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/56—Porous materials, e.g. foams or sponges
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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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/58—Materials at least partially resorbable by the body
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- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
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- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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Abstract
A kind of promotion, which is press-offed, injures the collagen material that avulsion skin survives, it is made of collagen protein sponge and nutrient solution, specifically, it is that ox heel string, pig heel string are passed through into collagen made from the smart extraction of enzymolysis, purification step for raw material, collagen protein sponge, and nutrient solution are formed after being crosslinked freeze-drying process.Nutrient solution is added while wound is implanted into.The present invention inserts conveniently, it is simple applied to surgical procedure, effectively facilitate to press-off and injure the skin of avulsion and survive, promote suffering limb function early recovery, reduce avulsion or avulsion injuries cutaneous necrosis rate, prevent the appearance of cutaneous necrosis and bone tendon exposed complication, thus caused by Repeated Operation the features such as, farthest reduces back the postoperative complication that plant is performed the operation or flap surgery etc. is brought；It is adapted in hand and foot surgery and four limbs avulsion injuries, avulsion, it is suitable to promote and apply.
Description
Technical field
It is more specifically, more particularly to a kind of to promote to press-off to injure avulsion skin the present invention relates to field of medical materials
The collagen material survived.
Background technology
Serious hand and foot and limbs skin avulsion injuries or avulsion, particularly retrograde avulsion or the treatment of avulsion injuries are orthopaedics
More intractable problem in processing.Returned in the past frequently with skin and plant operation or the operation of a variety of Vessel Skin Flaps.Skin returns plant art and often makes
Into cutaneous necrosis or partial necrosis, bone tendon exposed, needs again or Repeated Operation；Vessel Skin Flap is performed the operation, and operating difficulty is big, especially
It is free skin flap, and wound weight, risk are high, are suitable only for the hospital of microsurgical technique strength richness, and patient usually controls
Hospital stays length is treated, operation number is more, and flap has the complication such as downright bad probability or too fat to move, skin donor site damage, causes curative effect to owe
It is good.Skin for ease of avulsion injuries or avulsion survives, and operation is changed into simple and practicable, preferably promotes suffering limb functional rehabilitation,
There is an urgent need for a kind of collagen material to promote surviving for avulsion injuries and avulsion skin.
The content of the invention
(1) technical problem
Therefore, how to solve the problems, such as that avulsion injuries and avulsion skin survive, it is urgently to be resolved hurrily into those skilled in the art
The problem of.
(2) technical solution
Press-off in view of the above-mentioned problems, present disclosure is to provide a kind of promotion and injure the collagen egg that avulsion skin survives
White material.
A kind of promotion, which is press-offed, injures the collagen material that avulsion skin survives, and is by collagen protein sponge and nutrient solution
Composition, specifically, be by ox heel string, pig heel string for raw material by enzymolysis, purification step essence extraction made from collagen, pass through
Collagen protein sponge, and nutrient solution are formed after crosslinking freeze-drying process.Nutrient solution is added while wound is implanted into.
The collagen protein sponge is made through following steps：
1. pre-treatment：Ox heel string, pig heel string are rejected into shell, adipose tissue, muscle and other impurities is removed, uses tap water
Cleaning 5 times, is cleaned 5 times with deionized water, then is disinfected with 75% alcohol immersion 30min.The tissue for being cut into 1cm × 1cm is small
It is put into after block in high-speed tissue mashing machine and smashs into paste to pieces, rotating speed is 10000~12000r/min, then uses and contains 4.5mol/L
The 0.05mol/L Tris-HCl buffer solutions (pH 7.5) of NaCl are fully washed to remove the components such as lipid and serum, last low temperature
Centrifugation, centrifugal rotational speed 8000r/min, centrifugation time 30min, centrifuging temperature are 4 DEG C.
2. digest：Enzymolysis liquid is added, stirs and maintains 4 DEG C, 72h is extracted, is centrifuged after being sufficiently stirred with magnetic stirring apparatus,
Gained supernatant collagen stoste.
The enzymolysis liquid is the 0.5mol/L glacial acetic acid of the 500mg containing pepsin, and the dosage of the enzymolysis liquid is：Need to digest
Material：Enzymolysis liquid=1：10(V：V).
Centrifugal rotational speed is 4000r/min, and centrifugation time 20min, centrifuging temperature is 4 DEG C
3. purify：The NaCl solution that concentration is 4.4mol/ is added in collagen stoste, is stirred continuously until white wadding
Shape precipitation is separated out from solution, continuously adds NaCl solution until precipitating not untill precipitation, at 4 DEG C, the rotating speed of 5000r/min
White depositions are obtained after lower centrifugation 20min.Added in the white depositions molten in the acetum that concentration is 0.5mol/L
Solution.Dissolved collagen solution is continued to be repeated processing of saltouing, repeats this step 2~4 times.
4. it is crosslinked：Collagen after purification is obtained into collagen protein sponge with cross-linking agents 3h.The crosslinking agent uses
The dimethyl formamide solution of 0.30% DPPA (diphenyl phosphate azide).Using DPPA as collagen egg made from crosslinking agent
Bai Haimian has good biocompatibility and stability.
5. dry：Collagen protein sponge after coach is placed in vacuum freeze drier and carries out freeze-drying 48h, temperature
For -30~-40 DEG C, vacuum is 6~4Pa.
6. sterilize：Sterilized using co-60 radiation.
The nutrient solution is acidic growth factor, preferred acidic fibroblast growth factor.
The present invention mechanism be：(1) present invention is that collagen is carried out crosslinking Treatment, forms the glue with network
Former protein sponge, by means of the network structure of collagen protein sponge, serves as the supporting structure of subcutis.By collagen
While protein sponge is implanted into wound, nutrient solution is added, promotes local angeogenesis, makes the capillary for coming from the surface of a wound
With the component such as fibroblast, grow into an orderly manner in net structure；Meanwhile collagen protein sponge can be temporarily attached to bone, tendon
Transported Deng surface blood at difference or the tissue or organ transported almost without blood, for temporarily replacing the corium of defect；Have after (2) 2~3 weeks
Fibroblast and capillary are gradually degraded from surface of a wound mother bed and perienchyma's intrusion collagen protein sponge layer, collagen
And replaced by newborn granulation tissue, the presence of the collagen protein sponge because being attached to the surfaces such as bone, tendon, makes granulation tissue
It can be grown on wherein, be contacted so as to establish blood fortune with bone or tendon that the script of deep layer is transported without blood, the bone tendon for no blood fortune is created
Face skin-grafting provides condition；Meanwhile also be avulsion injuries, the blood of avulsion transport poor skin and improve blood fortune, promote its skin into
It is living.
A kind of promotion, which is press-offed, injures the collagen material that avulsion skin survives, and is that one kind is applied to avulsion injuries or avulsion
Hinder the material that skin survives, promote surviving for avulsion injuries or avulsion skin, prevent cutaneous necrosis and bone tendon exposed, promote to suffer from
Limb function early recovery, the auxiliary absorbable material for reducing complication.The collagen material is a kind of absorbable material, is utilized
It promotes the principle of blood capillary proliferation, and realization, which is press-offed, injures surviving for avulsion skin, while reduces cutaneous necrosis and bone flesh
The complication such as tendon is exposed.
(3) beneficial effect
The present invention inserts conveniently, simple applied to surgical procedure, effectively facilitates to press-off and injures the skin of avulsion and survive, and promotees
Into suffering limb function early recovery, avulsion or avulsion injuries cutaneous necrosis rate are reduced, prevents cutaneous necrosis and bone tendon exposed complication
Appearance so that caused by Repeated Operation the features such as, farthest reduces back the operation that plant is performed the operation or flap surgery etc. is brought
Complication；It is adapted in hand and foot surgery and four limbs avulsion injuries, avulsion, it is suitable to promote and apply.
Embodiment
With reference to embodiment, the invention will be further described.
A kind of promotion, which is press-offed, injures the collagen material that avulsion skin survives, and is by collagen protein sponge and nutrient solution
Composition, specifically, be by ox heel string, pig heel string for raw material by enzymolysis, purification step essence extraction made from collagen, pass through
Collagen protein sponge, and nutrient solution are formed after crosslinking freeze-drying process.Nutrient solution is added while wound is implanted into.
The collagen protein sponge is made through following steps：
1. pre-treatment：Ox heel string, pig heel string are rejected into shell, adipose tissue, muscle and other impurities is removed, uses tap water
Cleaning 5 times, is cleaned 5 times with deionized water, then is disinfected with 75% alcohol immersion 30min.The tissue for being cut into 1cm × 1cm is small
It is put into after block in high-speed tissue mashing machine and smashs into paste to pieces, rotating speed is 10000~12000r/min, then uses and contains 4.5mol/L
The 0.05mol/L Tris-HCl buffer solutions (pH 7.5) of NaCl are fully washed to remove the components such as lipid and serum, last low temperature
Centrifugation, centrifugal rotational speed 8000r/min, centrifugation time 30min, centrifuging temperature are 4 DEG C.
2. digest：Enzymolysis liquid is added, stirs and maintains 4 DEG C, 72h is extracted, is centrifuged after being sufficiently stirred with magnetic stirring apparatus,
Gained supernatant collagen stoste.
The enzymolysis liquid is the 0.5mol/L glacial acetic acid of the 500mg containing pepsin, and the dosage of the enzymolysis liquid is：Need to digest
Material：Enzymolysis liquid=1：10(V：V).
Centrifugal rotational speed is 4000r/min, and centrifugation time 20min, centrifuging temperature is 4 DEG C
3. purify：The NaCl solution that concentration is 4.4mol/ is added in collagen stoste, is stirred continuously until white wadding
Shape precipitation is separated out from solution, continuously adds NaCl solution until precipitating not untill precipitation, at 4 DEG C, the rotating speed of 5000r/min
White depositions are obtained after lower centrifugation 20min.Added in the white depositions molten in the acetum that concentration is 0.5mol/L
Solution.Dissolved collagen solution is continued to be repeated processing of saltouing, repeats this step 2~4 times.
4. it is crosslinked：Collagen after purification is obtained into collagen protein sponge with cross-linking agents 3h.The crosslinking agent uses
The dimethyl formamide solution of 0.30% DPPA (diphenyl phosphate azide).Using DPPA as collagen egg made from crosslinking agent
Bai Haimian has good biocompatibility and stability.
5. dry：Collagen protein sponge after coach is placed in vacuum freeze drier and carries out freeze-drying 48h, temperature
For -30~-40 DEG C, vacuum is 6~4Pa.
6. sterilize：Sterilized using co-60 radiation.
The nutrient solution is acidic growth factor, preferred acidic fibroblast growth factor.
The present invention is that collagen is carried out crosslinking Treatment, forms the collagen protein sponge with network, by means of
The network structure of collagen protein sponge, serves as the supporting structure of subcutis.Collagen protein sponge is implanted into wound
While, add nutrient solution, promote local angeogenesis, capillary and fibroblast for making to come from the surface of a wound etc. into
Point, grow into an orderly manner in net structure；Meanwhile to be temporarily attached to the surface such as bone, tendon blood fortune poor or several for collagen protein sponge
At tissue or organ without blood fortune, for temporarily replacing the corium of defect；There are fibroblast and blood capillary after 2~3 weeks
Pipe is gradually degraded from surface of a wound mother bed and perienchyma's intrusion collagen protein sponge layer, collagen and by newborn granulation tissue
Being replaced, the presence of the collagen protein sponge because being attached to the surfaces such as bone, tendon, enables granulation tissue to be grown on wherein, so that
The bone or tendon transported with the script of deep layer without blood are established blood fortune and are contacted, and condition is provided for the bone tendon surface of a wound skin-grafting of no blood fortune；
Meanwhile also be avulsion injuries, the blood of avulsion transport poor skin and improve blood fortune, promote its skin to survive.
Embodiment：Operating procedure using collagen material provided by the invention is：First, thorough debridement in operation,
Foreign matter is removed, retains the skin of avulsion injuries or avulsion, retains subcutaneous tendon and sclerotin, Iodophor, hydrogen peroxide and physiological saline
Irrigate repeatedly.Secondly, fracture row replacement and fixation, the tendon suture reparation of damage, above tendon and sclerotin, avulsion injuries skin
And the skin of avulsion transfers collagen material provided by the invention, the skin of avulsion injuries cuts osculum with sharp knife and is conducive to drain.
Finally, the skin wound of avulsion injuries or avulsion is covered with vacuum suction.
The present invention is not limited to the above-described embodiments, anyone can draw other various shapes under the enlightenment of the present invention
The product of formula.All equivalent changes and modifications done according to scope of the present invention patent, should all belong to the covering scope of the present invention.
Claims (3)
- It is by collagen protein sponge and nutrient solution group 1. a kind of promotion, which is press-offed, injures the collagen material that avulsion skin survives Into, it is characterised in that：It is that ox heel string, pig heel string are passed through into collagen made from the smart extraction of enzymolysis, purification step, warp for raw material Collagen protein sponge, and nutrient solution are formed after crossing crosslinking freeze-drying process.
- 2. a kind of promotion according to claim 1, which is press-offed, injures the collagen material that avulsion skin survives, its feature It is：The collagen protein sponge is made through following steps：1. pre-treatment：Ox heel string, pig heel string are rejected into shell, adipose tissue, muscle and other impurities is removed, is cleaned with tap water 5 times, cleaned 5 times with deionized water, then disinfected with 75% alcohol immersion 30min；After being cut into the tissue fritter of 1cm × 1cm It is put into high-speed tissue mashing machine and smashs into paste to pieces, rotating speed is 10000~12000r/min, then uses NaCl containing 4.5mol/L 0.05mol/L Tris-HCl buffer solutions (pH 7.5) fully washing to remove the components such as lipid and serum, last low temperature from The heart, centrifugal rotational speed 8000r/min, centrifugation time 30min, centrifuging temperature are 4 DEG C；2. digest：Enzymolysis liquid is added, stirs and maintains 4 DEG C, 72h is extracted, is centrifuged after being sufficiently stirred with magnetic stirring apparatus, gained Supernatant collagen stoste；The enzymolysis liquid is the 0.5mol/L glacial acetic acid of the 500mg containing pepsin, and the dosage of the enzymolysis liquid is：Need zymolyte Matter：Enzymolysis liquid=1：10(V：V)；Centrifugal rotational speed is 4000r/min, and centrifugation time 20min, centrifuging temperature is 4 DEG C；3. purify：The NaCl solution that concentration is 4.4mol/ is added in collagen stoste, is stirred continuously until that white flock sinks Shallow lake separated out from solution, continuously add NaCl solution until precipitate not untill precipitation, under 4 DEG C, the rotating speed of 5000r/min from White depositions are obtained after heart 20min；Add in the acetum that concentration is 0.5mol/L and dissolve in the white depositions；Will Dissolved collagen solution continues to be repeated processing of saltouing, and repeats this step 2~4 times；4. it is crosslinked：Collagen after purification is obtained into collagen protein sponge with cross-linking agents 3h, the crosslinking agent uses The dimethyl formamide solution of 0.30% DPPA (diphenyl phosphate azide)；5. dry：Collagen protein sponge after coach is placed in vacuum freeze drier and carries out freeze-drying 48h, temperature for- 30~-40 DEG C, vacuum is 6~4Pa；6. sterilize：Sterilized using co-60 radiation.
- 3. a kind of promotion according to claim 1, which is press-offed, injures the collagen material that avulsion skin survives, its feature It is：The nutrient solution is acidic growth factor, preferred acidic fibroblast growth factor.
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