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CN102475909A - Porous scaffold material for tissue engineering - Google Patents

Porous scaffold material for tissue engineering Download PDF

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Publication number
CN102475909A
CN102475909A CN2010105534416A CN201010553441A CN102475909A CN 102475909 A CN102475909 A CN 102475909A CN 2010105534416 A CN2010105534416 A CN 2010105534416A CN 201010553441 A CN201010553441 A CN 201010553441A CN 102475909 A CN102475909 A CN 102475909A
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CN
China
Prior art keywords
high polymer
gene recombination
porous support
spider
support
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Pending
Application number
CN2010105534416A
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Chinese (zh)
Inventor
王爽
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
DALIAN CHUANGDA TECHNOLOGY TRADE MARKET CO LTD
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DALIAN CHUANGDA TECHNOLOGY TRADE MARKET CO LTD
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Priority to CN2010105534416A priority Critical patent/CN102475909A/en
Publication of CN102475909A publication Critical patent/CN102475909A/en
Pending legal-status Critical Current

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Abstract

The invention belongs to the field of biomedical material scientific engineering, and relates to a porous scaffold material for tissue engineering. The tissue engineering scaffold material is obtained by blending the gene recombinant spider silk protein and the biodegradable high polymer, wherein the ratio (mass ratio) of the gene recombinant spider silk protein to the biodegradable high polymer is as follows: the porous scaffold material obtained by the invention has good biocompatibility, degradability and superior mechanical properties, and not only has simple processing technology and low cost, but also is suitable for industrialized mass production.

Description

A kind of used in tissue engineering porous support materials
Technical field
The invention belongs to field of biomedical materials, relate to a kind of tissue engineering rack material specifically.
Background technology
The depletion of tissue and organ, damage are topmost clinical medicine problems; And present Therapeutic Method mainly is organ transplantation, surgical repair, artificial substituent; Though these methods can play a role; But all there is such or such deficiency in they, are the method that cost " cures the wound with wound " like organ transplantation to sacrifice health tissues; There is the problem of biocompatibility in existing artificial substituent.Up to the eighties in 20th century; American scholar has proposed the organizational project regenerative medicine; It is principle and a method of utilizing life sciences and engineering science, and research and development is used for substituting tissue or the part of organ or substituent (Langer R, the Vacanti J P.Tissue engineering.Science of repertoire; 1993,260:920) 0 group
The development of weaver's journey has improved the depletion of tissue engineering tissue and organ, the treatment level of damage, has improved patient's quality of life, reduces medical treatment cost effectively.
One of main problem in science of tissue engineering research is to supply cell to carry out the interaction of timbering material and the cell and the timbering material of vital movement, and core is to set up the three dimensions complex that is made up of cell and biomaterial.Ideal biomaterial plays a crucial role in organizational project, and becomes the main flow of Tissue Engineering Study.
Experienced first generation inert material in the research of biomaterial, the second filial generation has activity or has after the degraded character, has developed into having degradable and bioactive third generation biomaterial (L.L.Hench etal. Science, 2002,295:1014) o concurrently
At present; Natural biologic material mainly is collagen protein in the organizational project; The substitute of synthetic mainly is that polylactic acid (PLA), polyglycolic acid (PGA) and polylactic acid and polyglycolic acid copolymer (PLGA) o will get ideal natural scaffold materials certain difficulty is arranged; And have some problems, strong like the antigenicity of collagen protein, mechanical strength is not enough, easy degeneration of collagen protein etc. in processing procedure; Though the substitute of synthetic also has biodegradable characteristics; Material implanted like U.S. FDA approval; Be made into absorbable suture, clamping plate, screw and dressing etc., but these materials need improve also at aspects such as the control of biocompatibility, physicochemical property, degradation rate and slow-releasing.Therefore, but at present the emphasis point of research more be to seek synthetic, good biocompatibility, degradable timbering material.
As the third generation biological material exploitation of organizational project, spider silk fibroin has advantageous condition.The Aranea cortina has the permeability of the good transparency, biodegradability and water one air interface.Similar with elastin laminin with collagen protein, spider silk fibroin tool self-assembly character is regulated so that mechanical support to be provided through secondary structure; Compare with polyester, the pliability and the elasticity of silk make it stand weight and fatigue.The fibroin good biocompatibility plays same cell adhesion, expansion, differentiation and growth with collagen.Silk substrate also has the mechanical induction effect, through the hardness of adjustment silk substrate, provides the final mechanical property of control substrate to imitate the mechanical property of natural body tissue and support growth in the host tissue.Therefore it shows great application potential (Winkler S on medical application, surgical sutures, biomaterial lining form and support, cell growth supporting bracket and sustained release substrate; Kaplan DL.Molecular biology of spidersilk. Reviews in Molecular Biotechnology; 2000,74:85).
The Li Min of Fujian Normal University etc. under the support of Fujian Province's natural science fund major scientific and technological projects (2001F 006), Ministry of Education's key project (02072) and project of national nature science fund project (30370414) respectively at (Li Min, Zhang Wenxian, Huang Jiankun etc. in 2002; The structure of spider dragline silk GFP and the expression in escherichia coli, biological engineering journal, 2002; 331) and (Li Min in 2004 18 (3):; Huang Jiankun, Tu Guiyun etc., structure, the expression and purification of RGD-spider dragline silk GFP; Biomedical engineering's magazine; 2004,21 (6): 1006) successively disclose the 26S Proteasome Structure and Function characteristic of this research group according to spider silk, using gene engineering is the modern biotechnology of core; Utilize the prokaryotic expression system; In conjunction with shot gun method and physical map spectrometry the full gene of spider dragline silk albumen is cloned, checked order and expresses, made up the spider's thread protein gene of multiple special sequence, but set up the technology such as fermentation tank high density fermentation process conditions of production; Make the public to have established the research and development basis for widely applying spider silk fibroin as the organizational project new material with simple and easy to do, low-cost and effective isolation and purification method scale preparation recombinant spider silk protein.
Summary of the invention
In order to solve the deficiency of existing tissue engineering material in biocompatibility, biological degradability and mechanical performance; The object of the invention be exactly according to the unique mechanical property of spider silk and biocompatibility with and essence be proteinic construction features, but the recombinant spider silk proteins and the biodegradable high polymer of disclosed scale preparation prepare good biocompatibility, degradable tissue engineering bracket material for primary raw material in utilization.This timbering material can not only be degraded to human body can absorb aminoacid, has good biocompatibility and mechanical performance preferably, and can produce in batches.
For realizing that the technical scheme that the object of the invention adopted is: with gene recombination spider silk fibroin and biodegradable high polymer is that primary raw material prepares good biocompatibility, degradable timbering material.Wherein the shared mass percent of gene recombination spider silk fibroin is 55-95%, and the shared mass percent of biodegradable high polymer is 45-50%.
Biodegradable high polymer is meant polylactic acid (PLA), polyglycolic acid (PGA), gathers breast among the present invention
Acid and polyglycolic acid copolymer (PLGA), paracyanogen for acrylic ester, polycaprolactone, gather the dioxa hexamethylene
Ketone and copolymer thereof, gather anhydride, poe, poly phosphazene, poly butyric, other linear aliphatic adoptions
Ester, polyamino acid, chitin, chitosan, cellulose, polyvinyl alcohol, in the polyoxyethylene a kind of or they
Combination.
The present invention can prepare porous support materials with the pore method, concrete technology:
1. the preparation support prepares liquid: get the gene recombination spider's thread protein powder and be dissolved in 80-98% formic acid and obtain egg
White solution adds high polymer again, and mixing promptly obtains containing the support preparation of gene recombination spider's thread protein and high polymer
Liquid.Wherein the shared mass percent of gene recombination spider's thread protein is 55-95%, and biodegradable height gathers
The shared mass percent of thing is 45-50%.
2. interpolation porogen: add porogen such as sodium chloride, mixing.
3. reverse mould, heating: the support that will be added with porogen prepares liquid to be poured in the mould county, places 55-90 ℃ of baking oven 10-60 minute formation support.
4. degeneration: support is immersed in taking-up after degeneration 10-15 hour in the denaturant solution (like ethanol),
Placed distilled water immersion 8-12 hours.
5. lyophilization: the support after the degeneration in-70 0C lyophilizations got final product in 1-5 hour the porous support material
Material.
The invention has the beneficial effects as follows:
1. the porous support materials that adopts gene recombination spider's thread protein to make is because its essence is natural egg
The white matter molecule makes it have good biocompatibility and degradability.
2. add biodegradable high polymer and not only further improve porous support mechanical performance, regulation and control bubble
The degradable characteristic of foam timbering material, and help reducing the cost of foam stand.
3. the processing technique of support is simple, and cost is low, also is fit to for industrialized mass.
Description of drawings
Fig. 1, Fig. 2 are foam stand material appearance figure.
Fig. 3, Fig. 4 are foam stand material profile sem photographs.
According to embodiment the present invention is further specified below.
The specific embodiment
EXAMPLE l
1. the preparation support prepares liquid: get 2. 25g gene recombination spider's thread protein powder and be dissolved in 7.5ml 98% formic acid and obtain 30% (w/v) protein solution, add the 0.45g polyvinyl alcohol again, and mixing, the support that promptly obtains containing gene recombination spider's thread protein and polyvinyl alcohol prepares liquid.
2. interpolation porogen: add 1. 5g porogen sodium chloride, mixing.
3. reverse mould, heating: the support that will be added with porogen prepares liquid to be poured in the cylindrical die (R=2cm), puts into 70 ℃ of baking ovens 30 minutes.
4. degeneration, distilled water immersion: support was immersed in the ethanol degeneration 15 hours, distilled water immersion 10 hours.
5. lyophilization: the support after the degeneration ,-70 ℃ got final product in freezing 2 hours porous support materials.
Embodiment 2
1. the preparation support prepares liquid: get 2. 5g gene recombination spider's thread protein powder and be dissolved in lOm1980h formic acid and obtain 25% (w/v) protein solution, add the 1.25g chitosan, mixing promptly obtains containing the gene recombination spider's thread egg
Support white and chitosan prepares liquid.
2. interpolation porogen: add 2.Og porogen sodium chloride, mixing.
3. reverse mould, heating: the support that will be added with porogen prepares liquid and pours rectangular die into
(4cmx3cmx3cm), put into 60 ℃ of baking ovens 20 minutes.
4. degeneration, distilled water immersion: support was immersed in the ethanol degeneration 10 hours, distilled water immersion 8 hours.
5. lyophilization: the support after the degeneration ,-70 ℃ got final product in freezing 4 hours porous support materials.
Embodiment 3
1. the preparation support prepares liquid: get 2g gene recombination spider's thread protein powder and be dissolved in 6m198% formic acid and obtain 33% (w/v) protein solution; Add 1.Og chitosan and 0.25g polyvinyl alcohol again; Mixing, the support that promptly obtains containing gene recombination spider's thread protein, chitosan and polyvinyl alcohol prepares liquid.
2. interpolation porogen: add 1. Og porogen sodium chloride, mixing.
3. reverse mould, heating: the support that will be added with porogen prepares liquid to be poured in the rectangular die (4cmx3cmx3cm), puts into 65 ℃ of baking ovens 30 minutes.
4. degeneration, distilled water immersion: support was immersed in the ethanol degeneration 13 hours, distilled water immersion 12 hours.
5. lyophilization: the support after the degeneration ,-70 ℃ got final product in freezing 3 hours porous support materials.

Claims (1)

1. the present invention relates to a kind of tissue engineered porous scaffold material, it is characterized in that obtaining porous support materials with gene recombination spider's thread protein and biodegradable high polymer blend,
(1). the protein of preparation porous support materials is gene recombination spider's thread protein;
(2). the high polymer of preparation porous support materials is biodegradable high polymer; Comprise polylactic acid (PLA), polyglycolic acid (PGA), polylactic acid and polyglycolic acid copolymer (PLGA), paracyanogen for acrylic ester, polycaprolactone, gather dioxanone and copolymer thereof, gather anhydride, poe, poly phosphazene, poly butyric, other linear aliphatic adoption esters, polyamino acid, chitin, chitosan, cellulose, polyvinyl alcohol, polyoxyethylene etc.
(3). the high polymer of preparation porous support materials can be wherein a kind of or their combination,
(4)., the proportioning of gene recombination spider's thread protein and biodegradable high polymer (mass ratio) is: the shared mass percent of gene recombination spider's thread protein is 55-95%, the shared mass percent of biodegradable high polymer is 45-50%.
CN2010105534416A 2010-11-22 2010-11-22 Porous scaffold material for tissue engineering Pending CN102475909A (en)

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Application Number Priority Date Filing Date Title
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Publications (1)

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CN102475909A true CN102475909A (en) 2012-05-30

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103865090A (en) * 2014-03-14 2014-06-18 山东理工大学 Method for improving hydrophilia of polypeptide film by using polycaprolactone and polyethylene glycol
CN105088779A (en) * 2015-08-05 2015-11-25 广州赛莱拉生物基因工程有限公司 Collagen attachment membrane and preparation method thereof
WO2020183465A1 (en) * 2019-03-11 2020-09-17 Seevix Material Sciences Ltd. Compositions comprising dragline spider silk
US12030919B2 (en) 2016-02-11 2024-07-09 Seevix Material Sciences Ltd. Composite materials comprising synthetic dragline spider silk
US12453688B2 (en) 2020-04-23 2025-10-28 Seevix Material Sciences Ltd. Cosmetic compositions comprising dragline spider silk

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103865090A (en) * 2014-03-14 2014-06-18 山东理工大学 Method for improving hydrophilia of polypeptide film by using polycaprolactone and polyethylene glycol
CN103865090B (en) * 2014-03-14 2016-04-27 山东理工大学 A kind of method that polycaprolactone and polyoxyethylene glycol improve poly-peptide film wetting ability
CN105088779A (en) * 2015-08-05 2015-11-25 广州赛莱拉生物基因工程有限公司 Collagen attachment membrane and preparation method thereof
CN105088779B (en) * 2015-08-05 2018-01-30 广州赛莱拉生物基因工程有限公司 A kind of collagem membrane patch and preparation method thereof
US12030919B2 (en) 2016-02-11 2024-07-09 Seevix Material Sciences Ltd. Composite materials comprising synthetic dragline spider silk
WO2020183465A1 (en) * 2019-03-11 2020-09-17 Seevix Material Sciences Ltd. Compositions comprising dragline spider silk
US12453688B2 (en) 2020-04-23 2025-10-28 Seevix Material Sciences Ltd. Cosmetic compositions comprising dragline spider silk

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Application publication date: 20120530