CN107441561A - A kind of preparation method of the bionical collagen-based material of active guide tissue regeneration reparation - Google Patents
A kind of preparation method of the bionical collagen-based material of active guide tissue regeneration reparation Download PDFInfo
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- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
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Abstract
The invention discloses a kind of preparation method of the bionical collagen-based material of active guide tissue regeneration reparation, comprise the following steps：1) low immunogenicity, high-purity biological active collagen, are prepared；2), adding acid anhydrides is acylated collagen；3), glycosaminoglycan is added to react to obtain the collagenglycosaminoglycan composite sponge with porous network structure；4), it is placed in EDC/NHS cross-linking systems and is crosslinked, the collagenglycosaminoglycan composite sponge being crosslinked；5), the collagenglycosaminoglycan composite sponge of crosslinking is immersed in growth factor solution, reaction obtains the bionical collagen-based material of active guide tissue regeneration reparation.The present invention solves the problem easily precipitated during collagenglycosaminoglycan compounding；Realize that material microcosmos network structure (pore size, porosity), mechanical strength are controllable, finally achieve the matching of angiogenesis speed and wound healing cycle and material property, growth factor actively guides wound repair and regeneration by delaying controlled release.
Description
Technical field
The present invention relates to a kind of wound repair material, specially a kind of bionical collagen-based of active guide tissue regeneration reparation
The preparation method of material.
Background technology
Wound repair material is to be used to post-operative wound nursing, burn, exterior trauma and long-term not more property ulcer wound repair
Multiple a kind of clinical medical material, mainly including traditional wound care products (such as cotton yarn bandage, dipping cotton yarn, cotton pad) and height
Hold wound repair material.Compared to traditional wound care products, high-end wound repair material is based on the theoretical exploitation of moist wounds healing,
It can improve speed of wound healing and healing quality, solve the problems such as chronic and complicated wound refractory conjunction, mitigation patient suffering, and
Replacement frequency is lower, more meets growth requirement and the mainstream technology direction of modern medicine.
With the development of social economy, aging population and thing followed diabetic ulcer, varicose ulcer, cotton-padded mattress
The increase of the chronic trauma patient such as sore, world market further expand to the demand of high-end wound repair material.According to statistics, 2012
Year high-end wound repair material market scale in the whole world accounts for the share of wound repair material market 50%, in advance in 7,500,000,000 U.S. dollars or so
Meter will increase to 15,200,000,000 dollars in 2016.Wherein, the U.S. occupies the market of high-end wound repair material about 47%, and Europe accounts for 34%,
Other countries and regions account for 19%, and high-end wound repair material market integrally presents a rapidly rising trend in American-European countries.At present, entirely
The main medical article manufacturer of 30, ball has developed more than 300 wound repair descriptions of materials, including a variety of biologies
The high-end products such as fundamental mode, non-adhesive type, special high-selenium corn type, skin substitutes type.
And domestic Wound care market is at present mainly based on traditional cotton care product.High-end wound repair material is only
4000000000 RMB or so, 20% market share is accounted for, and the enterprise for being engaged in high-end wound repair material production is no more than 10, product
In terms of performance and feature kind with external gap clearly.Patient is also remote low using the ratio of high-end wound repair product
In American-European countries, middle and high end field is almost that foreign brand name is monopolized.Generally speaking, China for high-end wound repair material still
In the stage of market cultivation, but according to the ASSOCIATE STATISTICS of 2011, by the 10% of national medical and health organization's surgery amount, live
20% meter of institute patient, there are about 1.1 hundred million person-times every year needs Wound care, and high-end Wound care market obviously has a high potential, in advance
The year two thousand twenty market scale is counted up to 20,000,000,000.
It is more that the high-end wound repair material in domestic market mainly includes collagen class, oxycellulose class, chitosan class and marine alga
The class product of carbohydrate etc. four.Wherein collagen class product is because with good biocompatibility, biodegradability, accelerating wound healing
The advantages that, turn into the product of current Clinical practice most main flow, be widely used in all kinds of Wound cares to guide wound repair, be adapted to
In almost each need to be using the chronic trauma patient of Advanced Nursing's pattern, acceleration encapsulation situations.Domestic market collagen-based
Wound repair product mainly includes skin Nike (import), times water chestnut, Ke Lao get, its victory, wound Fu Kang and can be nation etc., wherein skin Nike
The share in wound repair market nearly 90% is almost occupied, though home products has hemostasis, the function of repairing and fill simultaneously, is being faced
In default of product function subdivision and specialized guidance during bed application, more used as hemostasia products.
Simple collagen base biological material is intended only as support and passively directs wound repair, but can not active induced tissue
Regeneration.The function of material active guide tissue regeneration reparation can by it is compound it is a variety of have promote or suppress cell division breed,
Migrate with the tissue growth factor of gene expression effect to realize.The existing collagen base product in domestic market belongs to substantially passively to be drawn
Lead and repair this type.For high-end wound repair Material Field, simultaneously acceleration of wound reparation and tissue can be actively guided again
Raw product is only the final developing goal in future.The basic research that Domestic Scientific Research institutes is carried out to this at present is more, industrialization
With reference to defective tightness；And China's wound repair production scope of the enterprise is smaller, research strength is weak, and directly transition enters high-end
Wound repair market is also faced with no small difficulty, and the R ＆ D Industryization of active wound repair material quite lags at home.
The content of the invention
The technical problem to be solved in the present invention is to overcome existing single collagen-based materials, passively wound can only be promoted to repair
A kind of the defects of multiple, there is provided preparation method of the bionical collagen-based material of active guide tissue regeneration reparation.
In order to solve the above-mentioned technical problem, the invention provides following technical scheme：
A kind of preparation method of the bionical collagen-based material of active guide tissue regeneration reparation, comprises the following steps：
1) low immunogenicity, high-purity biological active collagen, are prepared；Preferably, the specific method of collagen is prepared with reference to specially
Profit number is ZL201110361034X patent of invention；
2) acid anhydrides, is added into collagen, at 4~15 DEG C, stirring, is acylated collagen；
3) glycosaminoglycan, is added to 2) middle, after mixing uniformly, centrifugation, bubble removing is removed, is then filled into mould, freezing is done
It is dry, obtain the collagen-glycosaminoglycan composite sponge with porous network structure；
4), collagen-glycosaminoglycan composite sponge is placed in EDC/NHS cross-linking systems and is crosslinked, is adjusted with L- polylysines
The degree of cross linking, reaction certain time is rocked at room temperature, then the crosslinking agent of distilled water cleaning removal residual, the collagen being crosslinked-
Glycosaminoglycan composite sponge；
5), the collagen-glycosaminoglycan composite sponge of crosslinking is immersed in growth factor solution, after adsorbing a period of time,
It is placed in mould, is freeze-dried, obtains the bionical collagen-based material of active guide tissue regeneration reparation.
Further, the mass concentration of collagen made from the step 1) is 0.5%~1.5%.
Further, the acid anhydrides in the step 2) is to appoint in acetic anhydride, propionic andydride, succinic anhydride and phthalic anhydride
Meaning is a kind of, and the amount for adding acid anhydrides is 5~20g/100g collagens.Preferably, the time of acylation reaction is 1~6h, and reaction pH is 7
~10.
Further, the glycosaminoglycan in the step 3) is hyaluronic acid, chondroitin sulfate, low molecular weight heparin, sulphur
The mass ratio of one or more in sour heparan and chitosan, glycosaminoglycan and collagen is 1:8~20.Preferably, it is transparent
The molecular weight of matter acid is 400~180kDa, and the molecular weight of chondroitin sulfate is 10~40kDa, the molecular weight of low molecular weight heparin
For 3000~8000Da, the molecular weight of Heparan sulfate is 20~50kDa.
Further, solvent is distilled water, 50% ethanol, 75% ethanol in EDC/NHS cross-linking systems in the step 4)
With one kind in 95% ethanol, EDC concentration is 0.5~10mmol/L, and NHS concentration is 0.125~2.5mmol/L, and L- gathers
The concentration of lysine is 0.1~5mmol/L.Preferably, the time of cross-linking reaction is 6~12h in step 4).
Further, the growth factor in the step 5) be bFGF, EGF, PDGF, CDGF and IGF-1 in one kind or
A variety of, the concentration of growth factor is 50~150 μ g/mL
Further, the thickness that the bionical collagen-based material of active guide tissue regeneration reparation is obtained in the step 5) is
1~5mm.
Prepare low immunogenicity, high-purity biological active collagen combines processing including pretreatment of raw material, grease removal, sour enzyme, surpassed
Sonicated, H2O2Solution processing, sour dissolved salt analysis purifying, dialysis purification, drying, are comprised the following steps that：(1) fresh animal group is taken
Knit (pigskin, ox-hide, ox heel string etc.), oil layer and trichocutis are removed in cutting, go the removal of impurity, are cleaned, by historrhexis into small
It is granular, according to solid-to-liquid ratio 1:30~1:40 ratio, 0.01~0.03mol/L sodium hydrate aqueous solution is added, in 6~8 DEG C
Immersion 1~2 hour, filtering are standby；(2) nothing that quality is 6~8 times of liver mass is added in above-mentioned pretreated tissue
Water ether or acetone, flowed back 5-8 hours in 35~40 DEG C, it is standby afterwards with distilled water flushing to free from extraneous odour；(3) will be above-mentioned
Tissue is immersed in the mixed liquor of 0.6~0.7mol/L glacial acetic acid and 600~700mg/L pepsins, and it is small persistently to stir 25~30
When；(4) gradual ultrasonication is used to said mixture, is first ultrasonically treated 30 minutes, reused using 100~200W
200~300W is ultrasonically treated 30 minutes, is finally ultrasonically treated 1 hour using 300~400W；(5) H2O2 of addition 2~3% is molten
Liquid, mixing stand 2~4 hours, adjust pH to 5.0, centrifugation；(6) supernatant is taken, adds NaCl, is stirred 20-30 hours, centrifugation,
Sediment adds the dissolving of 0.6-0.7moL/L glacial acetic acid, centrifugation, takes supernatant, adjusts pH to 7.5, adds NaCl, stirs 20-30
Hour, centrifugation；(7) sediment that step (6) obtains is dissolved in 0.6~0.7moL/L glacial acetic acid, first with 0.6~0.7moL/L
Glacial acetic acid for extracellular fluid dialysis dialyse 2 times, 4 hours every time, then using distilled water be extracellular fluid dialysis dialysis 5~7 times, 4 hours every time,
Chlorion is can't detect to outer liquid, obtains collagen liquid；(8) aforesaid liquid is freeze-dried, obtained with triple-helix structure work
The collagen of property.
The present invention by composite growth factor exploitation can active guide tissue regeneration reparation bionical collagen-based material.It is but raw
The long factor must could work with cells contacting more than certain time, and its half-life period is shorter, is easily easily degraded by proteases, and simply will
Growth factor can not be realized with collagen composite actively guides this function of wound repair.Creative introducing sugar in the present invention
Amine glycan, using the specific effect between glycosaminoglycan and growth factor, the slow controlled release that growth factor can be achieved is grown with adjusting
The activity of the factor, and then realize the active guide tissue regeneration repair function of collagen-based wound repair material.
The present invention uses bionical constructing technology, using low immunogenicity, high-purity biological active collagen as base material, passes through glue
Original-glycosaminoglycan is assembled into composite fibre altogether, solves the problem easily precipitated when the two is compounded, and then composite growth factor and real
The slow controlled release of existing growth factor, exploitation can active guide tissue regeneration reparation bionical collagen-based material.The product can be applied to
The treatment of various acute and chronic wounds, especially for the chronic wound that healing rate is slow, regeneration is difficult, vascularization degree is low,
It can reach the effect of accelerating wound repair and regeneration.The invention of the product, be expected to solve in wound repair field depth and
Vascularization degree existing for the clinic such as extensive wound, difficult healing wounds (such as diabetes, Venous Ulcers) is low, wound is cured
The problems such as sum velocity is slow, regeneration is difficult.
The modification that the present invention is acylated by collagen, the isoelectric point for making collagen is 4~4.5, solves collagen-glycosaminoglycan
The problem easily precipitated during compounding；By regulation of the L- polylysines to free amino group quantity in system, accurate control material
Crosslink density, the controllable adjustment in material degradation cycle is realized, make the pore size of material can be adjusted between 50~200 μm
Section, porosity can be adjusted between 80%~95%, and degradation cycle can be adjusted between 4~12 weeks, realize material
Microcosmos network structure (pore size, porosity), mechanical strength are controllable, finally achieve angiogenesis speed and wound healing week
Phase and the matching of material property.By collagen base biological material biomimetics constructing technology, composite growth factor and glycosaminoglycan, osamine
Glycan adjusts the slow release of growth factor while growth factor activity is protected, and growth factor is actively drawn by delaying controlled release
Lead wound repair and regeneration.
Brief description of the drawings
Accompanying drawing is used for providing a further understanding of the present invention, and a part for constitution instruction, the reality with the present invention
Apply example to be used to explain the present invention together, be not construed as limiting the invention.In the accompanying drawings：
Fig. 1 is collagen-based wound repair material microstructure SEM figures of the present invention.
Embodiment
The preferred embodiments of the present invention are illustrated below in conjunction with accompanying drawing, it will be appreciated that described herein preferred real
Apply example to be merely to illustrate and explain the present invention, be not intended to limit the present invention.
Embodiment 1
A kind of preparation method of the bionical collagen-based material of active guide tissue regeneration reparation, comprises the following steps：
1) low immunogenicity, high-purity biological active collagen that mass concentration is 0.5%, are prepared；
2) acetic anhydride, is added into collagen, at 4 DEG C, pH stirs for 7 times, acylation reaction 6h, and the addition of acetic anhydride is
5g/100g collagens；
3) it is, 180kDa hyaluronic acids to 2) the middle molecular weight that adds, after mixing uniformly, centrifugation, removes bubble removing, be then filled into
In mould, freeze-drying, the collagen-glycosaminoglycan composite sponge with porous network structure is obtained；Hyaluronic acid and collagen
Mass ratio is 1:8；
4), collagen-glycosaminoglycan composite sponge is placed in EDC/NHS cross-linking systems and is crosslinked, is adjusted with L- polylysines
The degree of cross linking, cross-linking reaction 6h is rocked at room temperature, then distilled water cleaning removes the crosslinking agent of residual, the collagen-sugar being crosslinked
Amine glycan composite sponge；Solvent is distilled water in EDC/NHS cross-linking systems, and EDC concentration is 0.5mmol/L, and NHS concentration is
The concentration of 0.125mmol/L, L- polylysine is 0.1mmol/L；
5) the collagen-glycosaminoglycan composite sponge of crosslinking, is immersed in the growth factor bFGF solution that concentration is 50 μ g/mL
In, after adsorbing a period of time, it is placed in mould, is freeze-dried, obtains the bionical collagen-based material of active guide tissue regeneration reparation
Material.
Embodiment 2
A kind of preparation method of the bionical collagen-based material of active guide tissue regeneration reparation, comprises the following steps：
1) low immunogenicity, high-purity biological active collagen that mass concentration is 1.5%, are prepared；
2) succinic anhydride, is added into collagen, at 15 DEG C, pH is 10 times stirring reaction 1h, is acylated collagen；Succinic acid
The addition of acid anhydride is 20g/100g collagens；
3) chondroitin sulfate that molecular weight is 40kDa, is added to 2) middle, after mixing uniformly, centrifugation, bubble removing is removed, then fills
Note in mould, be freeze-dried, obtain the collagen-glycosaminoglycan composite sponge with porous network structure；Chondroitin sulfate with
The mass ratio of collagen is 1:20；
4), collagen-glycosaminoglycan composite sponge is placed in EDC/NHS cross-linking systems and is crosslinked, is adjusted with L- polylysines
The degree of cross linking, reaction 12h is rocked at room temperature, then distilled water cleaning removes the crosslinking agent of residual, and the collagen-osamine being crosslinked gathers
Sugared composite sponge；Solvent is 50% ethanol in EDC/NHS cross-linking systems, and EDC concentration is 10mmol/L, and NHS concentration is
The concentration of 2.5mmol/L, L- polylysine is 5mmol/L；
5), by the collagen-glycosaminoglycan composite sponge of crosslinking be immersed in concentration be 150 μ g/mL growth factor PDGF it is molten
In liquid, after adsorbing a period of time, it is placed in mould, is freeze-dried, obtains the bionical collagen-based of active guide tissue regeneration reparation
Material.
Embodiment 3
A kind of preparation method of the bionical collagen-based material of active guide tissue regeneration reparation, comprises the following steps：
1) low immunogenicity, high-purity biological active collagen that mass concentration is 1.2%, are prepared；
2) phthalic anhydride, is added into collagen, at 10 DEG C, pH is 9 stirring reaction 3h, is acylated collagen；Add
The amount for entering acid anhydrides is 10g/100g collagens；
3) chitosan, is added to 2) middle, after mixing uniformly, centrifugation, bubble removing is removed, is then filled into mould, be freeze-dried,
Obtain the collagen-glycosaminoglycan composite sponge with porous network structure；The mass ratio of glycosaminoglycan and collagen is 1:14；
4), collagen-glycosaminoglycan composite sponge is placed in EDC/NHS cross-linking systems and is crosslinked, is adjusted with L- polylysines
The degree of cross linking, reaction 10h is rocked at room temperature, then distilled water cleaning removes the crosslinking agent of residual, and the collagen-osamine being crosslinked gathers
Sugared composite sponge；Solvent is 95% ethanol in EDC/NHS cross-linking systems, and EDC concentration is 5mmol/L, and NHS concentration is
The concentration of 1.8mmol/L, L- polylysine is 3mmol/L；
5), the collagen-glycosaminoglycan composite sponge of crosslinking is immersed in the growth factor solution that concentration is 100 μ g/mL,
After adsorbing a period of time, it is placed in mould, is freeze-dried, obtains the bionical collagen-based material of active guide tissue regeneration reparation.
Growth factor is the mixture in PDGF, CDGF and IGF-1.
The partial properties of the bionical collagen-based material of the active guide tissue regeneration reparation of the present invention of table 1
Pig full thickness dermal is repaiied using the bionical collagen-based material of the active guide tissue regeneration reparation of the present invention
It is multiple, the results showed that, collagen-based wound repair material structure is relatively compact, and porosity is high, is advantageous to migration and the nutrients of cell
The transport of matter, vascularization in 4 weeks is most obvious after graft materials, and collagen deposition is more, and fibrous connective tissue regeneration is good, and autologous skin is complete
Full survival, 6 weeks after stent graft, support is degraded substantially, corium sample tissue substitute stent implantation site, the degradation speed of support
Match with cambium formation speed, the reconstruction of dermal tissue with it is moulding can by good inductive formation, make skin graft effect it is good,
Meet clinically to the demand of dermal substitute.As a result collagen-based wound repair material produced by the present invention and import are shown
Skin Nike product is consistent or even excellent promote fibroblastic growth, connective tissue regeneration, tissue vascularization etc. the effect of
In skin Nike.
Finally it should be noted that：The preferred embodiments of the present invention are the foregoing is only, are not intended to limit the invention,
Although the present invention is described in detail with reference to the foregoing embodiments, for those skilled in the art, it still may be used
To be modified to the technical scheme described in foregoing embodiments, or equivalent substitution is carried out to which part technical characteristic.
Within the spirit and principles of the invention, any modification, equivalent substitution and improvements made etc., it should be included in the present invention's
Within protection domain.
Claims (10)
1. a kind of preparation method of the bionical collagen-based material of active guide tissue regeneration reparation, it is characterised in that including following
Step：
1) low immunogenicity, high-purity biological active collagen, are prepared；
2) acid anhydrides, is added into collagen, at 4~15 DEG C, stirring, is acylated collagen；
3) glycosaminoglycan, is added to 2) middle, after mixing uniformly, centrifugation, bubble removing is removed, is then filled into mould, be freeze-dried, obtain
To the collagen-glycosaminoglycan composite sponge with porous network structure；
4), collagen-glycosaminoglycan composite sponge is placed in EDC/NHS cross-linking systems and is crosslinked, is adjusted and is crosslinked with L- polylysines
Degree, reaction certain time is rocked at room temperature, then distilled water cleaning removes the crosslinking agent of residual, the collagen-osamine being crosslinked
Glycan composite sponge；
5), the collagen-glycosaminoglycan composite sponge of crosslinking is immersed in growth factor solution, after adsorbing a period of time, is placed in
In mould, freeze-drying, the bionical collagen-based material of active guide tissue regeneration reparation is obtained.
2. the preparation method of the bionical collagen-based material of active guide tissue regeneration reparation as claimed in claim 1, its feature
It is, the mass concentration of collagen made from the step 1) is 0.5%~1.5%.
3. the preparation method of the bionical collagen-based material of active guide tissue regeneration reparation as claimed in claim 1, its feature
It is, the acid anhydrides in the step 2) is any one in acetic anhydride, propionic andydride, succinic anhydride and phthalic anhydride, is added
The amount of acid anhydrides is 5~20g/100g collagens.
4. the preparation method of the bionical collagen-based material of active guide tissue regeneration reparation as claimed in claim 3, its feature
It is, the time of acylation reaction is 1~6h in the step 2), and reaction pH is 7~10.
5. the preparation method of the bionical collagen-based material of active guide tissue regeneration reparation as claimed in claim 1, its feature
Be, the glycosaminoglycan in the step 3) be hyaluronic acid, chondroitin sulfate, low molecular weight heparin, Heparan sulfate and
The mass ratio of one or more in chitosan, glycosaminoglycan and collagen is 1:8~20.
6. the preparation method of the bionical collagen-based material of active guide tissue regeneration reparation as claimed in claim 5, its feature
It is, the molecular weight of hyaluronic acid is 400~180kDa in the step 3), and the molecular weight of chondroitin sulfate is 10~40kDa,
The molecular weight of low molecular weight heparin is 3000~8000Da, and the molecular weight of Heparan sulfate is 20~50kDa.
7. the preparation method of the bionical collagen-based material of active guide tissue regeneration reparation as claimed in claim 1, its feature
It is, solvent is in distilled water, 50% ethanol, 75% ethanol and 95% ethanol in EDC/NHS cross-linking systems in the step 4)
One kind, EDC concentration is 0.5~10mmol/L, and NHS concentration is 0.125~2.5mmol/L, the concentration of L- polylysines
For 0.1~5mmol/L.
8. the preparation method of the bionical collagen-based material of active guide tissue regeneration reparation as claimed in claim 6, its feature
It is, the time of cross-linking reaction is 6~12h in the step 4).
9. the preparation method of the bionical collagen-based material of active guide tissue regeneration reparation as claimed in claim 6, its feature
Be, the growth factor in the step 5) be bFGF, EGF, PDGF, CDGF and IGF-1 in one or more, growth factor
Concentration be 50~150 μ g/mL.
10. the preparation method of the bionical collagen-based material of active guide tissue regeneration reparation as claimed in claim 6, its feature
It is, the thickness that the bionical collagen-based material of active guide tissue regeneration reparation is obtained in the step 5) is 1~5mm.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109045356A (en) * | 2018-09-08 | 2018-12-21 | 佛山市森昂生物科技有限公司 | A kind of preparation method of active biological film tissue mending material |
| WO2023125689A1 (en) * | 2021-12-28 | 2023-07-06 | Shanghai Qisheng Biological Preparation Co., Ltd. | Hyaluronic acid-collagen copolymer compositions and medical applications thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1788802A (en) * | 2005-12-14 | 2006-06-21 | 温州医学院 | Biological sponge material for promoting injury-restoring and its preparation method |
| CN101862475A (en) * | 2009-10-10 | 2010-10-20 | 广州市创伤外科研究所 | II type collagen hyaloplasm acid compound sponge bracket and use thereof |
-
2017
- 2017-06-16 CN CN201710466999.2A patent/CN107441561B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1788802A (en) * | 2005-12-14 | 2006-06-21 | 温州医学院 | Biological sponge material for promoting injury-restoring and its preparation method |
| CN101862475A (en) * | 2009-10-10 | 2010-10-20 | 广州市创伤外科研究所 | II type collagen hyaloplasm acid compound sponge bracket and use thereof |
Non-Patent Citations (1)
| Title |
|---|
| 李国英，刘文涛等: "《胶原化学》", 30 April 2013, 中国轻工业出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109045356A (en) * | 2018-09-08 | 2018-12-21 | 佛山市森昂生物科技有限公司 | A kind of preparation method of active biological film tissue mending material |
| WO2023125689A1 (en) * | 2021-12-28 | 2023-07-06 | Shanghai Qisheng Biological Preparation Co., Ltd. | Hyaluronic acid-collagen copolymer compositions and medical applications thereof |
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