CN107899080A - A kind of dermis restoration stent preparation method - Google Patents
A kind of dermis restoration stent preparation method Download PDFInfo
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- CN107899080A CN107899080A CN201711233172.3A CN201711233172A CN107899080A CN 107899080 A CN107899080 A CN 107899080A CN 201711233172 A CN201711233172 A CN 201711233172A CN 107899080 A CN107899080 A CN 107899080A
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- 210000004207 dermis Anatomy 0.000 title claims abstract description 57
- 238000002360 preparation method Methods 0.000 title claims abstract description 31
- 102000008186 Collagen Human genes 0.000 claims abstract description 41
- 108010035532 Collagen Proteins 0.000 claims abstract description 41
- 229920001436 collagen Polymers 0.000 claims abstract description 41
- 229920001287 Chondroitin sulfate Polymers 0.000 claims abstract description 27
- 229940059329 chondroitin sulfate Drugs 0.000 claims abstract description 27
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims abstract description 17
- SQDAZGGFXASXDW-UHFFFAOYSA-N 5-bromo-2-(trifluoromethoxy)pyridine Chemical compound FC(F)(F)OC1=CC=C(Br)C=N1 SQDAZGGFXASXDW-UHFFFAOYSA-N 0.000 claims abstract description 13
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 claims abstract description 6
- 150000001718 carbodiimides Chemical class 0.000 claims abstract description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 11
- 239000012528 membrane Substances 0.000 claims description 11
- 235000011194 food seasoning agent Nutrition 0.000 claims description 5
- 239000000017 hydrogel Substances 0.000 claims description 5
- 239000000377 silicon dioxide Substances 0.000 claims description 5
- 229910003460 diamond Inorganic materials 0.000 claims description 4
- 239000010432 diamond Substances 0.000 claims description 4
- 229920006264 polyurethane film Polymers 0.000 claims description 4
- 238000004080 punching Methods 0.000 claims description 3
- 238000005057 refrigeration Methods 0.000 claims description 3
- 230000008014 freezing Effects 0.000 claims 1
- 238000007710 freezing Methods 0.000 claims 1
- 238000009777 vacuum freeze-drying Methods 0.000 abstract description 3
- 239000010410 layer Substances 0.000 description 96
- 239000007788 liquid Substances 0.000 description 14
- 239000000047 product Substances 0.000 description 10
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 239000000463 material Substances 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 239000011241 protective layer Substances 0.000 description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 208000027418 Wounds and injury Diseases 0.000 description 6
- 238000001914 filtration Methods 0.000 description 6
- 239000000843 powder Substances 0.000 description 6
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 5
- 230000010261 cell growth Effects 0.000 description 5
- 239000012620 biological material Substances 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 239000008213 purified water Substances 0.000 description 4
- 102000057297 Pepsin A Human genes 0.000 description 3
- 108090000284 Pepsin A Proteins 0.000 description 3
- 238000004132 cross linking Methods 0.000 description 3
- 238000000502 dialysis Methods 0.000 description 3
- 238000007654 immersion Methods 0.000 description 3
- 239000012535 impurity Substances 0.000 description 3
- 239000011159 matrix material Substances 0.000 description 3
- 210000003205 muscle Anatomy 0.000 description 3
- 229940111202 pepsin Drugs 0.000 description 3
- 229920001606 poly(lactic acid-co-glycolic acid) Polymers 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 230000037314 wound repair Effects 0.000 description 3
- 229920002567 Chondroitin Polymers 0.000 description 2
- 102000012422 Collagen Type I Human genes 0.000 description 2
- 108010022452 Collagen Type I Proteins 0.000 description 2
- 230000032683 aging Effects 0.000 description 2
- DLGJWSVWTWEWBJ-HGGSSLSASA-N chondroitin Chemical compound CC(O)=N[C@@H]1[C@H](O)O[C@H](CO)[C@H](O)[C@@H]1OC1[C@H](O)[C@H](O)C=C(C(O)=O)O1 DLGJWSVWTWEWBJ-HGGSSLSASA-N 0.000 description 2
- 229940096422 collagen type i Drugs 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000002500 effect on skin Effects 0.000 description 2
- 239000003292 glue Substances 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 229920002521 macromolecule Polymers 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000002861 polymer material Substances 0.000 description 2
- 230000008929 regeneration Effects 0.000 description 2
- 238000011069 regeneration method Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 239000005864 Sulphur Substances 0.000 description 1
- 208000025865 Ulcer Diseases 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- -1 aperture 2mm Substances 0.000 description 1
- 230000008512 biological response Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000007541 cellular toxicity Effects 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 108010025899 gelatin film Proteins 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 239000005445 natural material Substances 0.000 description 1
- 231100001083 no cytotoxicity Toxicity 0.000 description 1
- 230000002028 premature Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 230000035882 stress Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000397 ulcer Toxicity 0.000 description 1
Classifications
-
- 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
- A61L27/14—Macromolecular materials
- A61L27/20—Polysaccharides
-
- 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
-
- 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
-
- 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
Abstract
The present invention discloses a kind of dermis restoration stent preparation method, first prepares mesh-supported layer and prepares and mandruka layer solution；Then mesh-supported layer is laid on lyophilized mold bottom, then collagen chondroitin sulfate solution is poured into mould, vacuum freeze drying, the mesh-supported layer and mandruka layer being be combined with each other；Then it is crosslinked in the glutaraldehyde, formaldehyde or the carbodiimides solution that are put in above-mentioned mesh-supported layer and mandruka layer, then cleans multipass, freeze again, finally obtain dermis restoration stent.The dermis restoration stent obtained by above-mentioned preparation method, has the characteristics that the shelf holding time is long, not degradable, good biocompatibility.
Description
Technical field
The present invention relates to a kind of dermis restoration stent preparation method, belong to medical dressing field.
Background technology
Full thickness dermal is clinically very common as caused by wound, burn and dermal chronic ulcer etc., how to realize
Effective wound repair and reconstruction are one of significant problems urgently to be resolved hurrily.Dermis restoration material is as induction corium regeneration
Template, the treatment for clinical deep dermis defect provides an ideal approach, but with natural biologic material, such as glue
Original etc., the dermis restoration material as matrix still suffer from the problem of regeneration induction repair ability in situ is insufficient.It is to find out its cause, main
For the stent of natural material, easily compressive deformation is difficult to the original three-dimensional porous structure of holding itself in mechanical environment in vivo and in vitro,
So that tissue, cell and growing into for blood vessel are restricted.Therefore, the machine of natural biologic material dermis restoration material how is improved
Tool supports problem, and carries out necessary functional modification to the dermis restoration material of structure on basis herein, makes it preferably
The biological response of excitation local wound microenvironment becomes the task of top priority.
At present, it is by the macromolecule such as PLGA, PGA or PLA braid and collagen to have some dermis restoration stent preparation methods
Matrix is integrated, but this kind of high molecular material of PLGA, PGA, PLA requires harshness to storage condition.Collagen under normal temperature condition
The moisture that matrix contains can cause product premature breakdown, so as to influence its shelf holding time, this kind of product normal temperature condition at present
It can only at most preserve 5 months.Such as needing to preserve for a long time needs in the environment for being deposited in less than 0 DEG C, and preservation condition is harsh, seriously
Constrain the application range of this kind of product.
In view of this, the present inventor studies this, specially develops a kind of dermis restoration stent preparation method, this case
Thus produce.
The content of the invention
The object of the present invention is to provide a kind of dermis restoration stent preparation method, can obtain a kind of shelf holding time it is long,
Not degradable, good biocompatibility dermis restoration stent.
To achieve these goals, solution of the invention is：
A kind of dermis restoration stent preparation method, includes the following steps：
It is prepared by mesh-supported layer：Mesh-supported layer is woven into using collagen line；Alternatively, mesh-supported is made using collagem membrane punching
Layer；
It is prepared by mandruka layer solution：The collagen solution and chondroitin sulfate solution that mass concentration is 0.2 ~ 2% are prepared respectively, will
Chondroitin sulfate solution is added dropwise in collagen solution, is stirred evenly, and obtains collagen-chondroitin sulfate solution, wherein chondroitin sulfate
Quality amount accounting is 1% ~ 50%；
Mesh-supported layer is laid on lyophilized mold bottom, then collagen-chondroitin sulfate solution is poured into mould, vacuum refrigeration is done
Mesh-supported layer and mandruka layer dry, be combined with each other；
Above-mentioned mesh-supported layer and mandruka layer are put in 0.01% ~ 2% glutaraldehyde, formaldehyde or carbodiimides solution and handed over
When connection 1 ~ 48 is small, then cleans 5 ~ 10 times repeatedly, freeze again, obtain dermis restoration stent.
Preferably, the dermis restoration stent preparation method is further comprising the steps of：
Silica hydrogel is applied on polyurethane film or pellosil, then the mandruka layer of dermis restoration stent is mutually glued with Silica hydrogel
Knot, obtains pad pasting type dermis restoration stent.
Preferably, before mesh-supported layer is positioned over lyophilized mould, first collagen-sulphur is poured into lyophilized mold bottom
Aching and limp ossein solution, freeze after formed mandruka layer, then mesh-supported layer is laid on mandruka layer, then by collagen-
Chondroitin sulfate solution is poured into mould, vacuum freeze drying, obtains mandruka layer, the mesh-supported layer mutually combined successively
With mandruka layer.
Preferably, the collagem membrane preparation process is：It is molten that collagen solution is added into glutaraldehyde, formaldehyde or carbodiimides
Liquid is crosslinked, and 25-40 DEG C of hot-air seasoning, is made collagem membrane.
Preferably, the collagem membrane preparation process is：Collagen solution is first passed through into 25-40 DEG C of hot-air seasoning, Ran Houyong
Ultraviolet lamp is crosslinked, and collagem membrane is made.
Preferably, the mesh-supported layer being prepared is equipped with multiple mesh.
Preferably, the mandruka layer being prepared is equipped with multiple micropores being mutually communicated.
Preferably, the mesh-supported layer mesh area is 0.0025 ~ 1cm2 。
Preferably, the mesh is circular, square, diamond shape, ellipse or polygon.
Preferably, the mandruka layer micropore size is 50 ~ 400 μm.
A kind of dermis restoration stent, mesh-supported layer and mandruka are obtained by above-mentioned dermis restoration stent preparation method
Layer is that collagen is made, and the ageing-resistant performance of collagen is better than other artificial synthesized degradable high polymer materials, because
Prepared by dermis restoration stent shelf life it is longer.Secondly, put by mesh-supported layer and mandruka stacking, three-dimensional structure
Stablize, mechanical strength is good, easy to operation stitching.And obtained dermis restoration stent component is simple, no toxic degradation product, carefully
Cellular toxicity is low, and biocompatibility is good, is conducive to wound repair.Thus, application and popularization more conducively clinically.
The present invention is described in further detail below in conjunction with drawings and the specific embodiments.
Brief description of the drawings
Fig. 1 is the dermis restoration stent diagrammatic cross-section of embodiment 1；
Fig. 2 is the mesh-supported layer segment structure diagram of embodiment 1；
Fig. 3 is the mandruka layer micro-structure diagram of embodiment 1；
Fig. 4 is the dermis restoration stent diagrammatic cross-section of embodiment 2；
Fig. 5 is the dermis restoration stent diagrammatic cross-section of embodiment 3.
Embodiment
Embodiment 1
A kind of preparation method of dermis restoration stent, includes the following steps：
It is prepared by collagen：Fresh ox heel string is taken, muscle shell and impurity is removed, cleans up, ox heel string is cut into 2 ~ 3mm using slicer
Thick sheet, according to solid-to-liquid ratio 1:50 ratio, adds the sodium hydroxide solution of 0.1mol/L, when 20 ~ 30 DEG C of immersions 10 are small,
Filtering, it is spare；By the above-mentioned ox heel string being obtained by filtration, constantly rinsed under the purified water of flowing, make its acid-base value 6 ~ 8 it
Between；Above-mentioned ox heel string is fully crushed in pulverizer；The ox heel string of crushing is immersed in the acetic acid enzymolysis liquid that pH is 2 ~ 3, its
Middle pepsin content is 500mg/L, when 2 ~ 8 DEG C of enzymolysis 24 are small；Above-mentioned enzymolysis liquid is put into centrifuge, in 5000r/
Min is centrifuged 10 minutes；Supernatant is taken, is slowly added into sodium chloride powder, is gently agitated for, collects product of saltouing；By above-mentioned salt division
Thing be fitted into bag filter it is every 3 it is small when change a water, when dialysis 24 is small, obtain collagen；
It is prepared by mesh-supported layer 1：The Collagen type-I line of a diameter of 0.20 ~ 0.249mm is taken, it is respectively 20cm's to be woven into length and width
Mesh-supported layer 1, mesh area are about 0.04cm2, 1 thickness of mesh-supported layer is about 0.4mm.
It is prepared by mandruka layer solution：Above-mentioned collagen is configured to the collagen solution that concentration is 1%, then presses collagen：Sulfuric acid
Chondroitin=99:1 ratio is slowly added into chondroitin sulfate powder under the conditions of 4 DEG C, maintains stirring in adition process, glue is made
Original-chondroitin sulfate liquid；
The lyophilized mold bottom of 20 × 20cm is laid in after the mesh-supported layer 1 woven by collagen line is soaked, then is poured into
Collagen-chondroitin sulfate liquid, solution height is controlled between 2 ~ 3mm, the netted branch that vacuum freeze drying is be combined with each other
Support layer 1 and mandruka layer 2；
Above-mentioned mesh-supported layer 1 and mandruka layer 2 are immersed in the glutaraldehyde solution that concentration is 1%, it is small in 4 DEG C of crosslinkings 12
When, then rinsed repeatedly in distilled water 5 times, be placed again into freeze dryer and carry out secondary freeze；
Above-mentioned lyophilized obtained dermis restoration stent is pasted into upper pellosil, then obtains pad pasting type dermis restoration stent.Silica gel
Film can protect the surface of a wound, prevent surface of a wound moisture evaporation.
A kind of dermis restoration stent obtained by above-mentioned preparation method, as shown in Figs. 1-3, including sets gradually netted
Supporting layer 1, mandruka layer 2 and protective layer 3, mandruka layer 2 is supported by mesh-supported layer 1.Wherein, the mesh-supported
Layer 1 is latticed, and equipped with multiple mesh 11, the mandruka layer 2 is equipped with multiple micropores 21, the setting of multiple micropores 21, and one
Aspect provides space for cell growth, on the other hand can make it have certain elasticity and hygroscopicity, and mandruka layer 2 prepares material
Expect including collagen, chondroitin sulfate isoreactivity biomaterial.The protective layer 3 is pellosil, is covered in mandruka layer 2
On.
1 mesh of mesh-supported layer is diamond shape, and cross-sectional area is about 0.04cm2, mesh can also be according to different process
It is set to circular, square, oval or polygon etc..21 aperture of mandruka layer micropore is 50 ~ 150 μm, the netted branch
The thickness for supportting layer 1 is about 0.4mm；The thickness of the mandruka layer 2 is 1mm.
The dermis restoration stent of said structure is first covered on human body wound in Clinical practice, passes through suture and wound
Surrounding tissue is in contact.The mesh-supported layer 1, protective layer 3 have certain degree of hardness compared with mandruka layer 2, in suture
It can promote the stress support at the same time of protective layer 3 and mesh-supported layer 1, so that structure is more after ensureing whole dermis restoration stent suture
Add stabilization, cambium can be promoted preferably to grow into mandruka layer 2.Mesh-supported layer 1 is made of collagen, phase
Compare the macromolecule braid such as traditional PLGA, PGA or PLA, the ageing-resistant performance of collagen is artificial synthesized better than other
Degradable high polymer material, shelf life is longer under the conditions of normal temperature storage, not degradable usually up to 3 ~ 4 years；And it is degraded
Product is mainly amino acid, can be absorbed by the body, no cytotoxicity, has good biocompatibility, is conducive to wound repair.
The mandruka layer 2 is made of collagen, chondroitin sulfate isoreactivity biomaterial, and is dispersed with and largely mutually passes through
Logical micropore 21, these micropores 21 provide the space of cell growth, and collagen isoreactivity material then provides cell growth
Nutriment.
Embodiment 2
A kind of preparation method of dermis restoration stent, includes the following steps：
It is prepared by collagen：Fresh ox heel string is taken, muscle shell and impurity is removed, cleans up, ox heel string is cut into 2 ~ 3mm using slicer
Thick sheet, according to solid-to-liquid ratio 1:50 ratio, adds the sodium hydroxide solution of 0.1mol/L, when 20 ~ 30 DEG C of immersions 10 are small,
Filtering, it is spare；By the above-mentioned ox heel string being obtained by filtration, constantly rinsed under the purified water of flowing, make its acid-base value 6 ~ 8 it
Between；Above-mentioned ox heel string is fully crushed in pulverizer；The ox heel string of crushing is immersed in the acetic acid enzymolysis liquid that pH is 2 ~ 3, its
Middle pepsin content is 500mg/L, when 2 ~ 8 DEG C of enzymolysis 24 are small；Above-mentioned enzymolysis liquid is put into centrifuge, in 5000r/
Min is centrifuged 10 minutes；Supernatant is taken, is slowly added into sodium chloride powder, is gently agitated for, collects product of saltouing；By above-mentioned salt division
Thing be fitted into bag filter it is every 3 it is small when change a water, when dialysis 24 is small, obtain collagen；
It is prepared by mesh-supported layer：Cut-off passes through the natural Collagen type-I line for 0.300 ~ 0.349mm, is woven into mesh-supported layer 1, its
Thickness is about 0.5mm, mesh area 0.08cm2；
It is prepared by mandruka layer solution：Above-mentioned collagen is configured to the collagen solution that concentration is 0.2%, then presses collagen：Sulfuric acid is soft
Ossein=10:1 ratio is slowly added into chondroitin sulfate powder under the conditions of 4 DEG C, maintains to stir in adition process, and obtained collagen-
Chondroitin sulfate liquid；
Be laid in the mold bottom of 20 × 20cm after the mesh-supported layer 1 woven by collagen line is soaked, then pour into collagen-
Chondroitin sulfate liquid, by solution height control between 2 ~ 3mm, freeze-drying obtains mesh-supported layer 1 and mandruka layer 2；
Above-mentioned mesh-supported layer 1 and mandruka layer 2 are immersed in the carbodiimides solution that concentration is 0.01%, in 4 DEG C of friendships
Join 48 it is small when, then rinsed repeatedly in distilled water 7 times, be placed again into freeze dryer carry out it is secondary lyophilized；
By above-mentioned lyophilized obtained dermis restoration stent progress cutting suitably, not pad pasting type dermis restoration stent is obtained.
A kind of dermis restoration stent obtained by above-mentioned preparation method, as shown in figure 4, including stacked mesh-supported layer
1 and mandruka layer 2,1 layer of the mesh-supported be located at 2 bottom of mandruka layer.The mesh-supported layer 1 is latticed, if
There are multiple mesh, the mandruka layer 2 is equipped with multiple micropores, and the setting of multiple micropores, on the one hand provides sky for cell growth
Between, certain elasticity and hygroscopicity on the other hand can be made it have, it is collagen that mandruka layer 2, which prepares main material,.
1 mesh of mesh-supported layer is square, and area is about 0.08cm2, mesh can also be circle, diamond shape, ellipse
Shape or polygon etc..The mandruka layer micropore size is about 150 ~ 250 μm, and the thickness of the mesh-supported layer 1 is
0.5mm；The thickness of the mandruka layer 2 is 5mm.
Embodiment 3
A kind of preparation method of dermis restoration stent, includes the following steps：
It is prepared by collagen：Fresh ox heel string is taken, muscle shell and impurity is removed, cleans up, ox heel string is cut into 2 ~ 3mm using slicer
Thick sheet.According to solid-to-liquid ratio 1:50 ratio, adds the sodium hydroxide solution of 0.1mol/L, when 20 ~ 30 DEG C of immersions 10 are small,
Filtering, it is spare；By the above-mentioned ox heel string being obtained by filtration, constantly rinsed under the purified water of flowing, make its acid-base value 6 ~ 8 it
Between；Above-mentioned ox heel string is fully crushed in pulverizer；The ox heel string of above-mentioned crushing is immersed into the acetic acid enzymolysis liquid that pH is 2 ~ 3
In, wherein pepsin content is 500mg/L, when 2 ~ 8 DEG C of enzymolysis 24 are small；Above-mentioned enzymolysis liquid is put into centrifuge, in
5000r/min is centrifuged 10 minutes；Supernatant is taken, is slowly added into sodium chloride powder, is gently agitated for, collects product of saltouing；Will be above-mentioned
Product of saltouing be fitted into bag filter it is every 3 it is small when change a water, when dialysis 24 is small, obtain collagen；
It is prepared by mesh-supported layer：Compound concentration is 1% collagen solution, is uniformly mixed under the conditions of 4 DEG C.Pour this solution into bottom
In the smooth stainless steel mould in face, highly in 3mm or so, the hot-air seasoning under the conditions of 30 DEG C, then with ultraviolet lamp crosslinking 4 it is small when.
Collagem membrane is taken out, using punching apparatus, is punched out on collagem membrane, aperture 2mm, pitch of holes 4mm, obtains mesh-supported
Layer 1；
It is prepared by mandruka layer solution：Above-mentioned collagen is configured to the collagen solution that concentration is 2%, then presses collagen：Chondroitin sulfate
Element=2:1 ratio is slowly added into chondroitin sulfate powder under the conditions of 4 DEG C, obtains collagen-chondroitin sulfate solution；
Above-mentioned collagen-chondroitin sulfate solution is cast in lyophilized mould, the height of cast is about 1mm.The mould is put
Enter -40 DEG C freeze 4 it is small when；
Mesh-supported layer 1 is laid on the above-mentioned collagen-chondroitin sulfate solution freezed, then pours into a mould upper collagen-sulfuric acid immediately
Chondroitin solution, is put into -40 DEG C and carries out secondary freeze；
Freeze dryer flaggy temperature is cooled to -20 DEG C in advance, above-mentioned lyophilized mould is put into freeze dryer is freezed, and obtains mandruka
Layer 2, mesh-supported layer 1, the dermis restoration stent of 2 three-decker of mandruka layer；The setting of double-layer porous spongy layer, can make
Mandruka layer and mesh-supported layer combination are closer, avoid in some cases mesh-supported layer separated with mandruka layer；
Above-mentioned dermis restoration stent is immersed in the formalin that concentration is 2%, when 4 DEG C of crosslinkings 24 are small, then in purified water
In rinse repeatedly 10 times, then product is put into freeze dryer again carry out it is secondary lyophilized；
Above-mentioned lyophilized obtained dermis restoration stent is pasted into upper polyurethane film by Silica hydrogel, both obtains pad pasting type dermis restoration branch
Frame.
A kind of dermis restoration stent obtained by the above method, as shown in figure 5, including the mandruka layer set gradually
2nd, mesh-supported layer 1, mandruka layer 2 and protective layer 3, the mesh-supported layer 1 is latticed, equipped with multiple mesh；It is described
Mandruka layer 2 is equipped with multiple micropores 21, and the setting of multiple micropores 21, on the one hand provides space for cell growth, on the other hand
Certain elasticity and hygroscopicity can be made it have, it is collagen that mandruka layer 2, which prepares main material,.The protective layer 3 covers
Wherein on a mandruka layer 2, protective layer 3 is polyurethane film.
1 mesh aperture of mesh-supported layer is 2mm, and mesh is circle.The mandruka layer micropore size for 200 ~
400 μm, the thickness of the mesh-supported layer 1 is about 300 μm；The thickness of the mandruka layer 2 is 3mm.
Above-described embodiment and schema and non-limiting product form of the invention and style, any technical field it is common
The appropriate change or modification that technical staff does it, all should be regarded as not departing from the patent category of the present invention.
Claims (10)
1. a kind of dermis restoration stent preparation method, it is characterised in that include the following steps：
It is prepared by mesh-supported layer：Mesh-supported layer is woven into using collagen line；Alternatively, mesh-supported is made using collagem membrane punching
Layer；
It is prepared by mandruka layer solution：The collagen solution and chondroitin sulfate solution that mass concentration is 0.2 ~ 2% are prepared respectively, will
Chondroitin sulfate solution is added dropwise in collagen solution, is stirred evenly, and obtains collagen-chondroitin sulfate solution, wherein chondroitin sulfate
Quality amount accounting is 1% ~ 50%；
Mesh-supported layer is laid on lyophilized mold bottom, then collagen-chondroitin sulfate solution is poured into mould, vacuum refrigeration is done
Mesh-supported layer and mandruka layer dry, be combined with each other；
Above-mentioned mesh-supported layer and mandruka layer are put in 0.01% ~ 2% glutaraldehyde, formaldehyde or carbodiimides solution and handed over
When connection 1 ~ 48 is small, then cleans 5 ~ 10 times repeatedly, freeze again, obtain dermis restoration stent.
A kind of 2. dermis restoration stent preparation method as claimed in claim 1, it is characterised in that：The dermis restoration stent system
Preparation Method is further comprising the steps of：
Silica hydrogel is applied on polyurethane film or pellosil, then the mandruka layer of dermis restoration stent is mutually glued with Silica hydrogel
Knot, obtains pad pasting type dermis restoration stent.
A kind of 3. dermis restoration stent preparation method as claimed in claim 1, it is characterised in that：Placed by mesh-supported layer
Before lyophilized mould, collagen-chondroitin sulfate solution first is poured into lyophilized mold bottom, mandruka layer is formed after freezing,
Then mesh-supported layer is laid on mandruka layer, then collagen-chondroitin sulfate solution is poured into mould, vacuum refrigeration is done
It is dry, obtain mandruka layer, mesh-supported layer and the mandruka layer mutually combined successively.
A kind of 4. dermis restoration stent preparation method as claimed in claim 1, it is characterised in that：The collagem membrane preparation process
For：Collagen solution is added glutaraldehyde, formaldehyde or carbodiimides solution to be crosslinked, 25-40 DEG C of hot-air seasoning, is made collagen
Film.
A kind of 5. dermis restoration stent preparation method as claimed in claim 1, it is characterised in that：The collagem membrane preparation process
For：Collagen solution is first passed through into 25-40 DEG C of hot-air seasoning, is then crosslinked with ultraviolet lamp, collagem membrane is made.
A kind of 6. dermis restoration stent preparation method as claimed in claim 1, it is characterised in that：The mesh-supported being prepared
Layer is equipped with multiple mesh.
A kind of 7. dermis restoration stent preparation method as claimed in claim 6, it is characterised in that：The mesh-supported layer mesh
Area is 0.0025 ~ 1cm2。
A kind of 8. dermis restoration stent preparation method as claimed in claim 6, it is characterised in that：The mesh is circular, side
Shape, diamond shape, ellipse or polygon.
A kind of 9. dermis restoration stent preparation method as claimed in claim 1, it is characterised in that：The mandruka being prepared
Layer is equipped with multiple micropores being mutually communicated.
A kind of 10. dermis restoration stent preparation method as claimed in claim 9, it is characterised in that：The mandruka layer is micro-
Hole aperture is 50 ~ 400 μm.
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