CN101524556A - Porous tissue engineering scaffold and preparation method thereof - Google Patents
Porous tissue engineering scaffold and preparation method thereof Download PDFInfo
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- CN101524556A CN101524556A CN 200910071752 CN200910071752A CN101524556A CN 101524556 A CN101524556 A CN 101524556A CN 200910071752 CN200910071752 CN 200910071752 CN 200910071752 A CN200910071752 A CN 200910071752A CN 101524556 A CN101524556 A CN 101524556A
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Abstract
The invention relates to a porous tissue engineering scaffold and a preparation method thereof. The invention solves the problem that the current tissue engineering scaffold has inferior compatibility with cells. The inventive porous tissue engineering scaffold is mainly prepared by a copolymer of alpha-hydroxy acid and amino acid and by a pore-foaming agent; the inventive method comprises the following steps: preparing the alpha-hydroxy acid and the amino acid into a polymer; molding or casting forming. The inventive porous tissue engineering scaffold has excellent compatibility with the cells. The inventive method is simple in process and convenient in operation.
Description
Technical field
The present invention relates to tissue engineering bracket and preparation method thereof.
Background technology
Tissue engineering bracket is one of important content of Tissue Engineering Study as the artificial extracellular matrix, and it has cell is delivered to intravital ad-hoc location, occupies certain space in advance for regenerated organizing then, and the effect of transmitting tissue's development.Present applied tissue engineering material mainly contains two big classes: natural biological macromolecular material and synthesizing polymeric material.Natural material costs an arm and a leg, the nature difference between the different batches material is big, and can't satisfy according to the organizational project needs performances such as degradation time are carried out designing requirement; Though synthetic polymer can improve these shortcomings of natural material, lack the cell recognition signal, lacking biological interaction with iuntercellular is that it is at the bottleneck of further using aspect the organizational project.
Polylactic acid and polyglycolic acid are through drugs approved by FDA, but also be the biomaterial of the vivo degradation of extensive use the earliest, these two kinds of materials can be degraded into harmless material in vivo fully, it is adjustable that these two kinds of materials also have degradation time simultaneously, characteristics such as physical and mechanical properties is good, thereby become the preferred material in present Tissue Engineering Study field, when but they carry out the cultivation of cell as tissue engineering bracket material, it is poor that the problem that at first runs into is exactly material and cell compatibility, be unfavorable for that cell adheres on material, sprawl, migration, propagation, thereby can't the large tracts of land promotion and application.
Summary of the invention
Technical problem to be solved by this invention is the problem of existing tissue engineering bracket and cell compatibility difference; And provide porous tissue engineering scaffold and preparation method thereof.
Porous tissue engineering scaffold of the present invention is mainly made by 'alpha '-hydroxy acids and amino acid whose copolymer and porogen, the porosity of porous tissue engineering scaffold is that 92.5%～98.5% (volume), aperture are 200～250 μ m, described 'alpha '-hydroxy acids is lactic acid and/or hydroxyacetic acid, aminoacid accounts for 2%～40% of copolymer gross mass, described porogen is sodium chloride or sucrose, and the mass ratio of porogen and copolymer is 4～9: 1.Also comprise active particle and/or somatomedin in the porous tissue engineering scaffold of the present invention; Wherein active particle accounts for 5%～30% of copolymer gross mass, and somatomedin accounts for 0.5%～10% of copolymer gross mass.In addition, can be at copolymer surface grafting polypeptide.
The present invention prepares the scheme one of porous tissue engineering scaffold, concrete preparation method is as follows: a, 'alpha '-hydroxy acids and aminoacid are carried out fusion polymerisation in bulk 24h under the catalysis of 110～130 ℃ and stannous octoate, use methanol extraction after being dissolved in chloroform then, under 35～45 ℃ of conditions, dry again, obtain copolymer; B, copolymer is dissolved in to be made into mass concentration in chloroform or the ether be 5%～30% solution, add porogen again, sonic oscillation waters after evenly and casts from the mold cavity, places 24h～48h at room temperature, then dry 24h～48h under 35～45 ℃ of conditions; Promptly making porosity is that 92.5%～98.5% (volume), aperture are the porous tissue engineering scaffold of 200～250 μ m; Wherein the described 'alpha '-hydroxy acids of step a is lactic acid and/or hydroxyacetic acid, and aminoacid accounts for 2%～40% of copolymer gross mass; The described porogen of step b is sodium chloride or sucrose, and the mass ratio of porogen and copolymer is 4～9: 1.
The step b of scheme one can also be undertaken by following step: copolymer is put into mold cavity, adding porogen again, is compression molding 10min～20min, cool to room temperature again under 8MPa～15MPa condition at 160～180 ℃, pressure then, with distilled water wash 4 to 8 times, lyophilizing gets final product again.
Scheme one in step b, add active particle and/or somatomedin again when adding porogen, wherein active particle accounts for 5%～30% of step a copolymer gross mass; Somatomedin accounts for 0.5%～10% of step a copolymer gross mass.
Also can be after the step b of scheme one by following step at copolymer surface grafting polypeptide, concrete operations are as follows: carry out ultraviolet disinfection earlier, put into PBS solution then, add cross-linking agent again, at room temperature place 12h～48h; Putting into concentration after the taking-up is the 1g/L polypeptide solution, at room temperature places 10h～15h and promptly finishes at copolymer surface grafting polypeptide; Wherein said cross-linking agent is Sulfo-LC-SPDP, and dosage of crosslinking agent is 3%～10% of a PBS solution quality, and described polypeptide solution is by polypeptide dry powder and the preparation of PBS solution.
The present invention prepares the scheme two of porous tissue engineering scaffold, concrete preparation method is as follows: one, 'alpha '-hydroxy acids is carried out fusion polymerisation in bulk 24h under 110～130 ℃, the catalysis of stannous octoate, use methanol extraction after being dissolved in chloroform then, oven dry obtains polymer under 35 ℃～45 ℃ conditions again; Two, the polymer that then step 1 is obtained, dicyclohexylcarbodiimide and N-hydroxyl-butanimide activate 1～9h by 1: 5: 5 mixed in molar ratio to polymer surfaces; Three, the aminoacid cyclisation is obtained polyamino acid, the polymer of polyamino acid and surface active is added in the dimethyl sulfoxine, the total polymer mass of polyamino acid and surface active and dimethyl sulfoxine mass ratio are 1～2: 6～5, place in the deionized water behind room temperature concussion 2～5h, mixing after-filtration, lyophilizing obtain copolymer (comb copolymer); Four, copolymer is dissolved in to be made into mass concentration in chloroform or the ether be 5%～30% solution, add porogen again, water after sonic oscillation is extremely even and cast from the mold cavity, place 24～48h in the room temperature, dry 24～48h under 35～45 ℃ of conditions then, obtaining porosity is that 92.5%～98.5% (volume), aperture are the porous tissue engineering scaffold of 200～250 μ m; Wherein the described 'alpha '-hydroxy acids of step 3 is lactic acid and/or hydroxyacetic acid, and aminoacid accounts for 2%～40% of copolymer gross mass; Porogen is sodium chloride or sucrose in the step 4, and the mass ratio of porogen and copolymer is 4～9: 1.
The step 4 of scheme two can also be undertaken by following step: copolymer is put into mold cavity, adding porogen again, is compression molding 10min～20min, cool to room temperature again under 8MPa～15MPa condition at 160～180 ℃, pressure then, with distilled water wash 4 to 8 times, lyophilizing gets final product again.
Scheme two in step 4, add active particle and/or somatomedin again when adding porogen, wherein active particle accounts for 5%～30% of step 3 copolymer gross mass; Somatomedin accounts for 0.5%～10% of step 3 copolymer gross mass.
Also can be after the step 4 of scheme two by following step at copolymer surface grafting polypeptide, concrete operations are as follows: carry out ultraviolet disinfection earlier, put into PBS solution then, add cross-linking agent again, at room temperature place 12h～48h; Putting into concentration after the taking-up is the 1g/L polypeptide solution, at room temperature places 10h～15h and promptly finishes at copolymer surface grafting polypeptide; Wherein said cross-linking agent is Sulfo-LC-SPDP, and dosage of crosslinking agent is 3%～10% of a PBS solution quality, and described polypeptide solution is by polypeptide dry powder and the preparation of PBS solution.
Above-mentioned active particle is a kind of or wherein several mixing in gelatin, collagen, glucosan, hyaluronic acid, chitin, chitosan, alginate, hydroxyapatite, the coral reef; Above-mentioned somatomedin is a kind of or wherein several mixing in skeletal growth factor, epidermal growth factor, insulin like growth factor, the nerve growth factor.Above-mentioned polypeptide is an arginine-glycine-aspartic acid.
The advantage that the present invention has:
(1) porous tissue engineering scaffold of the present invention contains the aminoacid unit, has excellent biological compatibility, no significant cytotoxicity, inflammatory reaction and immunologic rejection can not influence neoblastic function because of the rejection of adjacent tissue, help cell adhesion, sprawl, move, breed.
Porous tissue engineering scaffold of the present invention is implanted spinal cord half in the mice body of having no progeny, carry out structure observation after 30 days, transplanting place does not have obvious inflammatory reaction, tissue and cell grow fine, the cell compatibility that this shows porous tissue engineering scaffold of the present invention is good, and nerve growth is had tangible contact inducing action.
(2) porous tissue engineering scaffold of the present invention has degradability and suitable degradation rate, and when the cell or tissue of transplanting was survived in receptor, timbering material can be degraded voluntarily, and degraded and absorbed speed can be complementary with cell, tissue growth speed.
(3) porous tissue engineering scaffold of the present invention has suitable hole dimension, high porosity and the pore morphology that is connected, be beneficial to the drainage of transmission, metabolite of formation, oxygen and nutrition of growth, extracellular matrix of plantation, cell and the tissue of a large amount of cells and blood vessel and neural in grow into, adapt to sclerous tissues's cells such as osteocyte, chondrocyte to greatest extent, the purpose that various histiocytes such as endocrine tissue's cell such as pancreatic cell, thyroid cell and muscle, blood vessel, epithelium are cultivated.
(4) porous tissue engineering scaffold of the present invention has high surface area, suitable surface physicochemical property and good cell interface relation, is beneficial to cell adhesion, propagation, differentiation and bio signal molecule such as growth factor-loaded.
(5) structural strength that is complementary of the mechanical property of porous tissue engineering scaffold of the present invention and implant site tissue keeping structural stability and integrity in the biomechanics microenvironment in vivo, and improves suitable microstress environment for implanting cell.
(6) performance parameter of product of the present invention can design by at the requirement of human body different tissues cell to support according to cell, sneak into active particle, biotic factor or polypeptide, can in a big way, regulate its biocompatibility, mechanical property and intravital degradation property, make its requirement that more is applicable to tissue engineering bracket material, satisfy clinical medical application.
(7) technology of the present invention simple, be convenient to operation, processing easily, can be processed into ideal two dimension or three dimensional structure, can obtain required tissue or organ shape, be easy to repeat make, and can keep original shape after being transplanted in the body.
Description of drawings
Fig. 1 is that the porous tissue engineering scaffold of the specific embodiment 77 is implanted spinal cord half observation of the mouse tissue after 30 days photo in the mice body of having no progeny, and arrow refers to and implants the intravital porous tissue engineering scaffold of mice among the figure.
The specific embodiment
Technical solution of the present invention is not limited to the following cited specific embodiment, also comprises the combination in any between each specific embodiment.
The specific embodiment one: the present embodiment porous tissue engineering scaffold is made by 'alpha '-hydroxy acids and amino acid whose copolymer and porogen, the porosity of porous tissue engineering scaffold is that 92.5%～98.5% (volume), aperture are 200～250 μ m, described 'alpha '-hydroxy acids is lactic acid and/or hydroxyacetic acid, aminoacid accounts for 2%～40% of copolymer gross mass, all the other are 'alpha '-hydroxy acids, described porogen is sodium chloride or sucrose, and the mass ratio of porogen and copolymer is 4～9: 1.
When the present embodiment 'alpha '-hydroxy acids was mixture, lactic acid, hydroxyacetic acid were pressed arbitrarily than combination.The porous tissue engineering scaffold of present embodiment contains the aminoacid unit, has excellent biological compatibility, no significant cytotoxicity, inflammatory reaction and immunologic rejection, can not influence neoblastic function because of the rejection of adjacent tissue, help cell adhesion, sprawl, move, breed; And can degrade voluntarily, degraded and absorbed speed can be complementary with cell, tissue growth speed; And have suitable hole dimension, high porosity and a pore morphology that is connected, be beneficial to the drainage of transmission, metabolite of formation, oxygen and nutrition of growth, extracellular matrix of plantation, cell and the tissue of a large amount of cells and blood vessel and neural in grow into, adapt to sclerous tissues's cells such as osteocyte, chondrocyte to greatest extent, the purpose that various histiocytes such as endocrine tissue's cell such as pancreatic cell, thyroid cell and muscle, blood vessel, epithelium are cultivated.The present embodiment porous tissue engineering scaffold has high surface area, suitable surface physicochemical property and good cell interface relation, is beneficial to cell adhesion, propagation, differentiation and bio signal molecule such as growth factor-loaded.The structural strength that the mechanical property of the porous tissue engineering scaffold of present embodiment and implant site tissue is complementary keeping structural stability and integrity in the biomechanics microenvironment in vivo, and improves suitable microstress environment for implanting cell.
The specific embodiment two: what present embodiment and the specific embodiment one were different is: aminoacid accounts for 5%～35% of copolymer gross mass.Other is identical with the specific embodiment one.
The specific embodiment three: what present embodiment and the specific embodiment one were different is: aminoacid accounts for 10～30% of copolymer gross mass.Other is identical with the specific embodiment one.
The specific embodiment four: what present embodiment and the specific embodiment one were different is: aminoacid accounts for 15% of copolymer gross mass.Other is identical with the specific embodiment one.
The specific embodiment five: what present embodiment and the specific embodiment one were different is: aminoacid accounts for 20% of copolymer gross mass.Other is identical with the specific embodiment one.
The specific embodiment six: what present embodiment and the specific embodiment one were different is: aminoacid accounts for 25% of copolymer gross mass.Other is identical with the specific embodiment one.
The specific embodiment seven: what present embodiment and the specific embodiment one to six were different is: the copolymer surface also is grafted with polypeptide.Other is identical with the specific embodiment one to six.
The specific embodiment eight: what present embodiment and the specific embodiment seven were different is: polypeptide is an arginine-glycine-aspartic acid.Other is identical with the specific embodiment seven.
The specific embodiment nine: the porous tissue engineering scaffold of present embodiment is made by 'alpha '-hydroxy acids, amino acid whose copolymer, active particle and porogen, the porosity of porous tissue engineering scaffold is that 92.5%～98.5% (volume), aperture are 200～250 μ m, described 'alpha '-hydroxy acids is lactic acid and/or hydroxyacetic acid, aminoacid accounts for 2%～40% of copolymer gross mass, active particle accounts for 5%～30% of copolymer gross mass, all the other are 'alpha '-hydroxy acids, described porogen is sodium chloride or sucrose, and the mass ratio of porogen and copolymer is 4～9: 1.
When the present embodiment 'alpha '-hydroxy acids was mixture, lactic acid, hydroxyacetic acid were pressed arbitrarily than combination.
Present embodiment has enlarged the scope of biocompatibility, and mechanical property and intravital degradation property make it be applicable to the requirement of human body different tissues cell to support, satisfy clinical medical application.
The specific embodiment ten: what present embodiment and the specific embodiment nine were different is: aminoacid accounts for 5%～35% of copolymer gross mass.Other is identical with the specific embodiment nine.
The specific embodiment 11: what present embodiment and the specific embodiment nine were different is: aminoacid accounts for 10～30% of copolymer gross mass.Other is identical with the specific embodiment nine.
The specific embodiment 12: what present embodiment and the specific embodiment nine were different is: aminoacid accounts for 15% of copolymer gross mass.Other is identical with the specific embodiment nine.
The specific embodiment 13: what present embodiment and the specific embodiment nine were different is: aminoacid accounts for 20% of copolymer gross mass.Other is identical with the specific embodiment nine.
The specific embodiment 14: what present embodiment and the specific embodiment nine were different is: aminoacid accounts for 25% of copolymer gross mass.Other is identical with the specific embodiment nine.
The specific embodiment 15: what present embodiment and the specific embodiment nine to 14 were different is: active particle accounts for 10%～25% of copolymer gross mass.Other is identical with the specific embodiment nine to 14.
The specific embodiment 16: what present embodiment and the specific embodiment nine to 14 were different is: active particle accounts for 20% of copolymer.Other is identical with the specific embodiment nine to 14.
The specific embodiment 17: what present embodiment and the specific embodiment nine to 16 were different is: active particle is a kind of or wherein several mixing in gelatin, collagen, glucosan, hyaluronic acid, chitin, chitosan, alginate, hydroxyapatite, the coral reef.Other is identical with the specific embodiment nine to 16.
When active particle is mixture in the present embodiment, press between various active particles arbitrarily than mixing.
The specific embodiment 18: what present embodiment and the specific embodiment nine to 17 were different is: the copolymer surface grafting polypeptide that adds active particle.Other is identical with the specific embodiment nine to 17.
Present embodiment porous engineering rack strengthens the adhesion of cell, has enlarged the scope of biocompatibility, and mechanical property and intravital degradation property make it be applicable to the requirement of human body different tissues cell to support, satisfy clinical medical application.
The specific embodiment 19: what present embodiment and the specific embodiment 18 were different is: polypeptide is an arginine-glycine-aspartic acid.Other is identical with the specific embodiment 18.
The specific embodiment 20: the porous tissue engineering scaffold of present embodiment is made by 'alpha '-hydroxy acids, amino acid whose copolymer, somatomedin and porogen, the porosity of porous tissue engineering scaffold is that 92.5%～98.5% (volume), aperture are 200～250 μ m, described 'alpha '-hydroxy acids is lactic acid and/or hydroxyacetic acid, aminoacid accounts for 2%～40% of copolymer gross mass, somatomedin accounts for 0.5%～10% of copolymer gross mass, all the other are 'alpha '-hydroxy acids, described porogen is sodium chloride or sucrose, and the mass ratio of porogen and copolymer is 4～9: 1.
When the present embodiment 'alpha '-hydroxy acids was mixture, lactic acid, hydroxyacetic acid were pressed arbitrarily than combination.
Porous organization's support of present embodiment promotes the selectively acting between cell and the biomaterial, enlarged the scope of biocompatibility, mechanical property and intravital degradation property make it be applicable to the requirement of human body different tissues cell to support, satisfy clinical medical application.
Body embodiment 21: what present embodiment and the specific embodiment 20 were different is: aminoacid accounts for 5%～35% of copolymer gross mass.Other is identical with the specific embodiment 20.
The specific embodiment 22: what present embodiment and the specific embodiment 20 were different is: aminoacid accounts for 10～30% of copolymer gross mass.Other is identical with the specific embodiment 20.
The specific embodiment 23: what present embodiment and the specific embodiment 20 were different is: aminoacid accounts for 15% of copolymer gross mass.Other is identical with the specific embodiment 20.
The specific embodiment 24: what present embodiment and the specific embodiment 20 were different is: aminoacid accounts for 20% of copolymer gross mass.Other is identical with the specific embodiment 20.
The specific embodiment 25: what present embodiment and the specific embodiment 20 were different is: aminoacid accounts for 25% of copolymer gross mass.Other is identical with the specific embodiment 20.
The specific embodiment 26: what present embodiment and the specific embodiment 20 to 25 were different is: somatomedin accounts for 2%～8% of copolymer gross mass.Other is identical with the specific embodiment 20 to 25.
The specific embodiment 27: what present embodiment and the specific embodiment 20 to 25 were different is: somatomedin accounts for 5% of copolymer gross mass.Other is identical with the specific embodiment 20 to 25.
The specific embodiment 28: what present embodiment and the specific embodiment 20 to 27 were different is: somatomedin is a kind of or wherein several mixing in skeletal growth factor, epidermal growth factor, insulin like growth factor, the nerve growth factor.Other is identical with the specific embodiment 20 to 27.
When somatomedin was mixture in the present embodiment, the requirement by the organism different tissues between various somatomedin mixed.
The specific embodiment 29: what present embodiment and the specific embodiment 20 to 27 were different is: the copolymer surface grafting polypeptide that adds somatomedin.Other is identical with the specific embodiment 20 to 27.
The specific embodiment 30: what present embodiment and the specific embodiment 29 were different is: polypeptide is an arginine-glycine-aspartic acid.Other is identical with the specific embodiment 29.
Specific embodiment hentriaconta-: the porous tissue engineering scaffold of present embodiment is by 'alpha '-hydroxy acids, amino acid whose copolymer, active particle, somatomedin and porogen are made, the porosity of porous tissue engineering scaffold is 92.5%～98.5% (volume), the aperture is 200～250 μ m, described 'alpha '-hydroxy acids is lactic acid and/or hydroxyacetic acid, aminoacid accounts for 2%～40% of copolymer gross mass, active particle accounts for 5%～30% of copolymer gross mass, somatomedin accounts for 0.5%～10% of copolymer gross mass, all the other are 'alpha '-hydroxy acids, described porogen is sodium chloride or sucrose, and the mass ratio of porogen and copolymer is 4～9: 1.
When the present embodiment 'alpha '-hydroxy acids was mixture, lactic acid, hydroxyacetic acid were pressed arbitrarily than combination.
Present embodiment has enlarged the scope of biocompatibility, and mechanical property and intravital degradation property make it be applicable to the requirement of human body different tissues cell to support, satisfy clinical medical application.
The specific embodiment 32: what present embodiment was different with specific embodiment hentriaconta-is: aminoacid accounts for 5%～35% of copolymer gross mass.Other is identical with specific embodiment hentriaconta-.
The specific embodiment 33: what present embodiment was different with specific embodiment hentriaconta-is: aminoacid accounts for 10～30% of copolymer gross mass.Other is identical with specific embodiment hentriaconta-.
The specific embodiment 34: what present embodiment was different with specific embodiment hentriaconta-is: aminoacid accounts for 15% of copolymer gross mass.Other is identical with specific embodiment hentriaconta-.
The specific embodiment 35: what present embodiment was different with specific embodiment hentriaconta-is: aminoacid accounts for 20% of copolymer gross mass.Other is identical with specific embodiment hentriaconta-.
The specific embodiment 36: what present embodiment was different with specific embodiment hentriaconta-is: aminoacid accounts for 25% of copolymer gross mass.Other is identical with specific embodiment hentriaconta-.
The specific embodiment 37: what present embodiment and specific embodiment hentriaconta-to three 16 were different is: active particle accounts for 10%～25% of copolymer gross mass.Other is identical with specific embodiment hentriaconta-to three 16.
The specific embodiment 38: what present embodiment and specific embodiment hentriaconta-to three 16 were different is: active particle accounts for 20% of copolymer gross mass.Other is identical with specific embodiment hentriaconta-to three 16.
The specific embodiment 39: what present embodiment and specific embodiment hentriaconta-to three 18 were different is: somatomedin accounts for 2%～8% of copolymer gross mass.Other is identical with specific embodiment hentriaconta-to three 18.
The specific embodiment 40: what present embodiment and specific embodiment hentriaconta-to three 18 were different is: somatomedin accounts for 5% of copolymer gross mass.Other is identical with specific embodiment hentriaconta-to three 18.
The specific embodiment 41: what present embodiment and specific embodiment hentriaconta-to four ten were different is: active particle is a kind of or wherein several mixing in gelatin, collagen, glucosan, hyaluronic acid, chitin, chitosan, alginate, hydroxyapatite, the coral reef.Other is identical with specific embodiment hentriaconta-to four ten.
When active particle is mixture in the present embodiment, press between various active particles arbitrarily than mixing.
The specific embodiment 42: what present embodiment and specific embodiment hentriaconta-to four 11 were different is: somatomedin is a kind of or wherein several mixing in skeletal growth factor, epidermal growth factor, insulin like growth factor, the nerve growth factor.Other is identical with specific embodiment hentriaconta-to four 11.
When somatomedin was mixture in the present embodiment, the requirement by the organism different tissues between various somatomedin mixed.
The specific embodiment 43: what present embodiment and specific embodiment hentriaconta-to four 12 were different is: the copolymer surface grafting polypeptide that adds active ion and somatomedin.Other is identical with specific embodiment hentriaconta-to four 12.
The specific embodiment 44: what present embodiment and the specific embodiment 43 were different is: polypeptide is an arginine-glycine-aspartic acid.Other is identical with the specific embodiment 43.
The specific embodiment 45: the preparation method of porous tissue engineering scaffold is as follows in the present embodiment: a, 'alpha '-hydroxy acids and aminoacid are carried out fusion polymerisation in bulk 24h under the catalysis of 110～130 ℃ and stannous octoate, use methanol extraction after being dissolved in chloroform then, under 35～45 ℃ of conditions, dry again, obtain copolymer; B, copolymer is dissolved in to be made into mass concentration in chloroform or the ether be 5%～30% solution, add porogen again, sonic oscillation waters after evenly and casts from the mold cavity, places 24h～48h at room temperature, then dry 24h～48h under 35～45 ℃ of conditions; Promptly making porosity is that 92.5%～98.5% (volume), aperture are the porous tissue engineering scaffold of 200～250 μ m; Wherein the described 'alpha '-hydroxy acids of step a is lactic acid and/or hydroxyacetic acid, and aminoacid accounts for 2%～40% of copolymer gross mass; The described porogen of step b is sodium chloride or sucrose, and the mass ratio of porogen and copolymer is 4～9: 1.
Present embodiment stannous octoate consumption is 2～6% of 'alpha '-hydroxy acids and an aminoacid gross mass.
When the 'alpha '-hydroxy acids among the present embodiment step a was mixture, lactic acid, hydroxyacetic acid were pressed arbitrarily than combination.The porous tissue engineering scaffold that present embodiment makes with ultraviolet disinfection after sealing preserve.
The porous tissue engineering scaffold of present embodiment contains the aminoacid unit, has excellent biological compatibility, no significant cytotoxicity, inflammatory reaction and immunologic rejection, can not influence neoblastic function because of the rejection of adjacent tissue, help cell adhesion, sprawl, move, breed; And can degrade voluntarily, degraded and absorbed speed can be complementary with cell, tissue growth speed.
The specific embodiment 46: what present embodiment and the specific embodiment 45 were different is: aminoacid accounts for 5%～35% of copolymer gross mass among the step a.Other step and parameter are identical with the specific embodiment 45.
The specific embodiment 47: what present embodiment and the specific embodiment 45 were different is: aminoacid accounts for 10～30% of copolymer gross mass among the step a.Other step and parameter are identical with the specific embodiment 45.
The specific embodiment 48: what present embodiment and the specific embodiment 45 were different is: aminoacid accounts for 15% of copolymer gross mass among the step a.Other step and parameter are identical with the specific embodiment 45.
The specific embodiment 49: what present embodiment and the specific embodiment 45 were different is: aminoacid accounts for 20% of copolymer gross mass among the step a.Other step and parameter are identical with the specific embodiment 45.
The specific embodiment 50: what present embodiment and the specific embodiment 45 were different is: aminoacid accounts for 25% of copolymer gross mass among the step a.Other step and parameter are identical with the specific embodiment 45.
The specific embodiment 51: what present embodiment and the specific embodiment 45 to 50 were different is: step b can also be undertaken by following step: copolymer is put into mold cavity, add porogen again, be compression molding 10min～20min under 8MPa～15MPa condition at 160～180 ℃, pressure then, cool to room temperature again, with distilled water wash 4 to 8 times, lyophilizing again; Promptly making porosity is that 92.5%～98.5% (volume), aperture are the porous tissue engineering scaffold of 200～250 μ m.Other step and parameter are identical with the specific embodiment 45 to 50.
The lyophilizing of present embodiment step 3 is to carry out freeze dried with liquid nitrogen.
The specific embodiment 52: what present embodiment and the specific embodiment 45 to 51 were different is: among the step b, add active particle and/or somatomedin when adding porogen again, wherein active particle accounts for 5%～30% of step a copolymer gross mass, and somatomedin accounts for 0.5%～10% of step a copolymer gross mass.Other step and parameter are identical with the specific embodiment 45 to 51.
The specific embodiment 53: what present embodiment and the specific embodiment 52 were different is: active particle accounts for 10%～25% of step a copolymer gross mass.Other step and parameter are identical with the specific embodiment 52.
The specific embodiment 54: what present embodiment and the specific embodiment 52 were different is: active particle accounts for 20% of step a copolymer gross mass.Other step and parameter are identical with the specific embodiment 52.
The specific embodiment 55: what present embodiment and the specific embodiment 52 to 54 were different is: somatomedin accounts for 2%～8% of step a copolymer gross mass.Other step and parameter are identical with the specific embodiment 52 to 54.
The specific embodiment 56: what present embodiment and the specific embodiment 52 to 54 were different is: somatomedin accounts for 5% of step a copolymer gross mass.Other step and parameter are identical with the specific embodiment 52 to 54.
The specific embodiment 57: what present embodiment and the specific embodiment 52 to 56 were different is: described active particle is a kind of or wherein several mixing in gelatin, collagen, glucosan, hyaluronic acid, chitin, chitosan, alginate microsphere, hydroxyapatite, the coral reef.Other step and parameter are identical with the specific embodiment 52 to 56.
When active particle is mixture in the present embodiment, press between various active particles arbitrarily than mixing.Alginate microsphere is a calcium alginate microsphere.
The specific embodiment 58: what present embodiment and the specific embodiment 52 to 57 were different is: described somatomedin is a kind of or wherein several mixing in skeletal growth factor, epidermal growth factor, insulin like growth factor, the nerve growth factor.Other step and parameter are identical with the specific embodiment 52 to 57.
When somatomedin was mixture in the present embodiment, the requirement by the organism different tissues between various somatomedin mixed.
The specific embodiment 59: what present embodiment and the specific embodiment 45 to 58 were different is: also can be by following step at copolymer surface grafting polypeptide after step b, concrete operations are as follows: carry out ultraviolet disinfection earlier, put into PBS solution then, add cross-linking agent again, at room temperature place 12h～48h; Putting into concentration after the taking-up is the 1g/L polypeptide solution, at room temperature places 10h～15h and promptly finishes at copolymer surface grafting polypeptide; Wherein said cross-linking agent is a Sulfo-LC-SPDP (sulfosuccinic acyl-2-pyridine two sulfur-3, propionamido 6-caproate, Sulfosuccinimidyl6-(3 '-[2-pyridyldithio]-propionamido) hexanoate), dosage of crosslinking agent is 3%～10% of a PBS solution quality, and described polypeptide solution is by polypeptide dry powder and the preparation of PBS solution.Other step and parameter are identical with the specific embodiment 45 to 58.
The compound method of the described PBS solution of present embodiment (phosphate buffered solution) is as follows: dissolving 8g NaCl, 0.2g KCl, 1.44g Na in the 800ml distilled water
2HPO
4With 0.24g KH
2PO
4, add water with HCl adjusting pH value to 7.4 and be settled to 1L, 1034 * 10
5Steam sterilization 20min under the Pa, room temperature preservation.
The specific embodiment 60: present embodiment with the specific embodiment 59 for different is: polypeptide is an arginine-glycine-aspartic acid in the described polypeptide solution.Other step and parameter are identical with the specific embodiment 59.
The specific embodiment 61: the preparation method of porous tissue engineering scaffold is as follows in the present embodiment: one, 'alpha '-hydroxy acids is carried out fusion polymerisation in bulk 24h under 110～130 ℃, the catalysis of stannous octoate, use methanol extraction after being dissolved in chloroform then, oven dry obtains polymer under 35 ℃～45 ℃ conditions again; Two, the polymer that then step 1 is obtained, dicyclohexylcarbodiimide and N-hydroxyl-butanimide activate 1～9h by 1: 5: 5 mixed in molar ratio to polymer surfaces; Three, the aminoacid cyclisation is prepared polyamino acid, the polymer of polyamino acid and surface active is added in the dimethyl sulfoxine, the total polymer mass of polyamino acid and surface active and dimethyl sulfoxine mass ratio are 1～2: 6～5, place in the deionized water behind room temperature concussion 2～5h, mixing after-filtration, lyophilizing obtain copolymer (comb copolymer); Four, copolymer is dissolved in to be made into mass concentration in chloroform or the ether be 5%～30% solution, add porogen again, water after sonic oscillation is extremely even and cast from the mold cavity, place 24～48h in the room temperature, dry 24～48h under 35～45 ℃ of conditions then, obtaining porosity is that 92.5%～98.5% (volume), aperture are the porous tissue engineering scaffold of 200～250 μ m; Wherein the described 'alpha '-hydroxy acids of step 3 is lactic acid and/or hydroxyacetic acid, and aminoacid accounts for 2%～40% of copolymer gross mass; Porogen is sodium chloride or sucrose in the step 4, and the mass ratio of porogen and copolymer is 4～9: 1.
Present embodiment stannous octoate consumption is 2～6% of 'alpha '-hydroxy acids and an aminoacid gross mass.The present embodiment step 3 is to adopt existing cyclization process that aminoacid is made polyamino acid.
When the 'alpha '-hydroxy acids in the present embodiment step 1 was mixture, lactic acid, hydroxyacetic acid were pressed arbitrarily than combination.The porous tissue engineering scaffold that present embodiment makes with ultraviolet disinfection after sealing preserve.
The porous tissue engineering scaffold of present embodiment contains the aminoacid unit, has excellent biological compatibility, no significant cytotoxicity, inflammatory reaction and immunologic rejection, can not influence neoblastic function because of the rejection of adjacent tissue, help cell adhesion, sprawl, move, breed; And can degrade voluntarily, degraded and absorbed speed can be complementary with cell, tissue growth speed.
The specific embodiment 62: what present embodiment and the specific embodiment 61 were different is: aminoacid accounts for 5%～35% of copolymer gross mass in the step 3.Other step and parameter are identical with the specific embodiment 61.
The specific embodiment 63: what present embodiment and the specific embodiment 61 were different is: aminoacid accounts for 10%～30% of copolymer gross mass in the step 3.Other step and parameter are identical with the specific embodiment 61.
The specific embodiment 64: what present embodiment and the specific embodiment 61 were different is: aminoacid accounts for 15% of copolymer gross mass in the step 3.Other step and parameter are identical with the specific embodiment 61.
The specific embodiment 65: what present embodiment and the specific embodiment 61 were different is: aminoacid accounts for 20% of copolymer gross mass in the step 3.Other step and parameter are identical with the specific embodiment 34.
The specific embodiment 66: what present embodiment and the specific embodiment 61 were different is: aminoacid accounts for 25% of copolymer gross mass in the step 3.Other step and parameter are identical with the specific embodiment 61.
The specific embodiment 67: what present embodiment and the specific embodiment 61 to 66 were different is: step 4 can also be undertaken by following step: copolymer is put into mold cavity, add porogen again, be compression molding 10min～20min under 8MPa～15MPa condition at 160～180 ℃, pressure then, cool to room temperature again, with distilled water wash 4 to 8 times, lyophilizing again; Promptly making porosity is that 92.5%～98.5% (volume), aperture are the porous tissue engineering scaffold of 200～250 μ m.Other step and parameter are identical with the specific embodiment 61 to 66.
The lyophilizing of present embodiment step 3 is to carry out freeze dried with liquid nitrogen.
The specific embodiment 68: what present embodiment and the specific embodiment 61 to four or six ten seven were different is: in the step 4, add active particle and/or somatomedin when adding porogen again, wherein active particle accounts for 5%～30% of step 3 copolymer gross mass, and somatomedin accounts for 0.5%～10% of step 3 copolymer gross mass.。Other step and parameter are identical with the specific embodiment 61 to four or six ten seven.
The specific embodiment 69: what present embodiment and the specific embodiment 68 were different is: active particle accounts for 10%～25% of step 3 copolymer gross mass.Other step and parameter are identical with the specific embodiment 68.
The specific embodiment 70: what present embodiment and the specific embodiment 68 were different is: active particle accounts for 20% of step 3 copolymer gross mass.Other step and parameter are identical with the specific embodiment 68.
The specific embodiment 71: what present embodiment and the specific embodiment 68 to 70 were different is: somatomedin accounts for 2%～8% of step 3 copolymer gross mass.Other step and parameter are identical with the specific embodiment 68 to 70.
The specific embodiment 72: what present embodiment and the specific embodiment 68 to 70 were different is: somatomedin accounts for 5% of step 3 copolymer gross mass.Other step and parameter are identical with the specific embodiment 68 to 70.
The specific embodiment 73: what present embodiment and the specific embodiment 68 to 72 were different is: described active particle is a kind of or wherein several mixing in gelatin, collagen, glucosan, hyaluronic acid, chitin, chitosan, alginate microsphere, hydroxyapatite, the coral reef.Other step and parameter are identical with the specific embodiment 68 to 72.
When active particle is mixture in the present embodiment, press between various active particles arbitrarily than mixing.The described alginate microsphere of present embodiment is a calcium alginate microsphere.
The specific embodiment 74: what present embodiment and the specific embodiment 68 to 73 were different is: described somatomedin is a kind of or wherein several mixing in skeletal growth factor, epidermal growth factor, insulin like growth factor, the nerve growth factor.Other step and parameter are identical with the specific embodiment 68 to 73.
When somatomedin was mixture in the present embodiment, the requirement by the organism different tissues between various somatomedin mixed.
The specific embodiment 75: what present embodiment and the specific embodiment 61 to 74 were different is: also can be by following step at copolymer surface grafting polypeptide after step b, concrete operations are as follows: carry out ultraviolet disinfection earlier, put into PBS solution then, add cross-linking agent again, at room temperature place 12h～48h; Putting into concentration after the taking-up is the 1g/L polypeptide solution, at room temperature places 10h～15h and promptly finishes at copolymer surface grafting polypeptide; Wherein said cross-linking agent is a Sulfo-LC-SPDP (sulfosuccinic acyl-2-pyridine two sulfur-3, propionamido 6-caproate, Sulfosuccinimidyl6-(3 '-[2-pyridyldithio]-propionamido) hexanoate), dosage of crosslinking agent is 3%～10% of a PBS solution quality, and described polypeptide solution is by polypeptide dry powder and the preparation of PBS solution.Other step and parameter are identical with the specific embodiment 61 to 74.
The compound method of the described PBS solution of present embodiment (phosphate buffered solution) is as follows: dissolving 8g NaCl, 0.2g KCl, 1.44g Na in the 800ml distilled water
2HPO
4With 0.24g KH
2PO
4, add water with HCl adjusting pH value to 7.4 and be settled to 1L, 1034 * 10
5Steam sterilization 20min under the Pa, room temperature preservation.
The specific embodiment 76: present embodiment with the specific embodiment 75 for different is: polypeptide is an arginine-glycine-aspartic acid in the described polypeptide solution.Other step and parameter are identical with the specific embodiment 75.
The specific embodiment 77: the preparation method of porous tissue engineering scaffold is as follows in the present embodiment: one, 'alpha '-hydroxy acids is carried out fusion polymerisation in bulk 24h under 130 ℃, the catalysis of stannous octoate, use methanol extraction (removal catalyst) after being dissolved in chloroform then, oven dry obtains polymer under 40 ℃ of conditions again; Two, the polymer that then step 1 is obtained, dicyclohexylcarbodiimide and N-hydroxyl-butanimide activate 2h by 1: 5: 5 mixed in molar ratio to polymer surfaces; Three, the lysine cyclisation is obtained polylysine, the polymer of polylysine and surface active is added in the dimethyl sulfoxine, the total polymer mass of polylysine and surface active and dimethyl sulfoxine mass ratio are 1～2: 6～5, place in the deionized water behind room temperature concussion 2～5h, mixing filters, and lyophilizing obtains copolymer (comb copolymer); Four, copolymer is put into mold cavity, add porogen again, compression molding 10min under 60～180 ℃, 10MPa condition, cool to room temperature then, with distilled water wash 5 times (removal porogen), the lyophilizing of reuse liquid nitrogen; Promptly obtaining porosity is that 92.5%～98.5% (volume), aperture are the porous tissue engineering scaffold of 200～250 μ m; Wherein the described 'alpha '-hydroxy acids of step 1 is a hydroxyacetic acid, and step 3 aminoacid accounts for 7% of copolymer gross mass; Porogen is a sodium chloride in the step 4, and the mass ratio of porogen and copolymer is 4～9: 1.
The comb copolymer weight average molecular weight that the present embodiment step 3 obtains is 50,000-100,000.
The porous tissue engineering scaffold of present embodiment is implanted spinal cord half in the mice body of having no progeny, carry out the structure observation (see figure 1) after 30 days, transplanting place does not have obvious inflammatory reaction, tissue and cell grow fine, the cell compatibility that this shows porous tissue engineering scaffold of the present invention is good, and nerve growth is had tangible contact inducing action.
The specific embodiment 78: what present embodiment and the specific embodiment 77 were different is: 'alpha '-hydroxy acids is a lactic acid in the step 1.Other step and parameter are identical with the specific embodiment 77.
The specific embodiment 79: what present embodiment and the specific embodiment 77 were different is: 'alpha '-hydroxy acids is the mixture of lactic acid and hydroxyacetic acid in the step 1, and lactic acid and hydroxyacetic acid mol ratio are 8: 2.Other step and parameter are identical with the specific embodiment 77.
The specific embodiment 80: what present embodiment and the specific embodiment 77 were different is: step 4 is finished by following reaction: copolymer is dissolved in to be made into mass concentration in the chloroform be 5%～30% solution, add NaCl again, water after sonic oscillation is extremely even and cast from the column type mould, placed 24 hours in the room temperature, under 40 ℃ of conditions dry 24 hours then, obtaining porosity was that 92.5%～98.5% (volume), aperture are the porous tissue engineering scaffold of 200～250 μ m.Other step and parameter are identical with the specific embodiment 77.
The specific embodiment 81: what this enforcement and the specific embodiment 80 were different is: adopt the lamellar mould in the step 4.Other step and parameter are identical with the specific embodiment 80.
The specific embodiment 82: what present embodiment and the specific embodiment 80 were different is: also can be by following step at the copolymer grafted polypeptide after step 4: carry out ultraviolet disinfection earlier, put into PBS solution then, add cross-linking agent again, at room temperature place 12～48h; Putting into concentration after the taking-up is the 1g/L polypeptide solution, at room temperature places 12～48h and promptly finishes at copolymer surface grafting polypeptide; Wherein said cross-linking agent is a Sulfo-LC-SPDP (sulfosuccinic acyl-2-pyridine two sulfur-3, propionamido 6-caproate, Sulfosuccinimidyl 6-(3 '-[2-pyridyldithio]-propionamido) hexanoate), dosage of crosslinking agent is 3%～10% of a PBS solution quality, and described polypeptide solution is by arginine-glycine-aspartic acid (RGD) dry powder and the preparation of PBS solution.Other step and parameter are identical with the specific embodiment 80.
The grafted RGD of present embodiment can promote receptor-mediated cell by the combination of intrinsic adhesion protein receptor-specific, in the spontaneous formation monolayer of biomaterial surface, for providing the site with receptor-mediated cell, and then promotes sticking, stretching of cell.
The specific embodiment 83: present embodiment and the specific embodiment 82 are different: step 4 is to be undertaken by following step: copolymer is put into mold cavity, add porogen again, compression molding 10min under 60～180 ℃, 10MPa condition, cool to room temperature then, with distilled water wash 5 times (removal porogen), lyophilizing again.Other step and parameter are identical with the specific embodiment 82.
The specific embodiment 84: what present embodiment and the specific embodiment 83 were different is: 'alpha '-hydroxy acids is a lactic acid in the step 1.Other step and parameter are identical with the specific embodiment 83.
The specific embodiment 85: what present embodiment and the specific embodiment 80 were different is: in the step 4, also add the calcium alginate microsphere that particle diameter is 100 μ m～250 μ m when adding porogen, calcium alginate microsphere accounts for 20% of step 3 copolymer gross mass.Other step and parameter are identical with the specific embodiment 80.
The specific embodiment 86: what present embodiment and the specific embodiment 84 were different is: ether substitutes chloroform in the step 4, also add calcium alginate microsphere and the skeletal growth factor that particle diameter is 100 μ m～250 μ m when adding porogen, calcium alginate microsphere accounts for 20% of step 3 copolymer gross mass, and skeletal growth factor accounts for 5% of step 3 copolymer gross mass.Other step and parameter are identical with the specific embodiment 84.
Claims (10)
1, porous tissue engineering scaffold, it is characterized in that porous tissue engineering scaffold mainly made by 'alpha '-hydroxy acids and amino acid whose copolymer and porogen, the porosity of porous tissue engineering scaffold is that 92.5%～98.5% (volume), aperture are 200～250 μ m, described 'alpha '-hydroxy acids is lactic acid and/or hydroxyacetic acid, aminoacid accounts for 2%～40% of copolymer gross mass, described porogen is sodium chloride or sucrose, and the mass ratio of porogen and copolymer is 4～9: 1.
2, porous tissue engineering scaffold according to claim 1 is characterized in that porous tissue engineering scaffold also comprises active particle and/or somatomedin; Wherein active particle accounts for 5%～30% of copolymer gross mass, and described active particle is a kind of or wherein several mixing in gelatin, collagen, glucosan, hyaluronic acid, chitin, chitosan, alginate, hydroxyapatite, the coral reef; Somatomedin accounts for 0.5%～10% of copolymer gross mass, and described somatomedin is a kind of or wherein several mixing in skeletal growth factor, epidermal growth factor, insulin like growth factor, the nerve growth factor.
3, porous tissue engineering scaffold according to claim 1 and 2 is characterized in that the copolymer surface is grafted with polypeptide.
4 porous tissue engineering scaffolds according to claim 3 is characterized in that described polypeptide is an arginine-glycine-aspartic acid.
5, the preparation method of porous tissue engineering scaffold, the preparation method that it is characterized in that porous tissue engineering scaffold is as follows: a, 'alpha '-hydroxy acids and aminoacid are carried out fusion polymerisation in bulk 24h under the catalysis of 110～130 ℃ and stannous octoate, use methanol extraction after being dissolved in chloroform then, under 35～45 ℃ of conditions, dry again, obtain copolymer; B, copolymer is dissolved in to be made into mass concentration in chloroform or the ether be 5%～30% solution, add porogen again, sonic oscillation waters after evenly and casts from the mold cavity, places 24h～48h at room temperature, then dry 24h～48h under 35～45 ℃ of conditions; Promptly making porosity is that 92.5%～98.5% (volume), aperture are the porous tissue engineering scaffold of 200～250 μ m; Wherein the described 'alpha '-hydroxy acids of step a is lactic acid and/or hydroxyacetic acid, and aminoacid accounts for 2%～40% of copolymer; The described porogen of step b is sodium chloride or sucrose, and the mass ratio of porogen and copolymer is 4～9: 1.
6, the preparation method of porous tissue engineering scaffold according to claim 5, it is characterized in that step b can also be undertaken by following step: copolymer is put into mold cavity, add porogen again, be compression molding 10min～20min under 8MPa～15MPa condition at 160～180 ℃, pressure then, cool to room temperature again, with distilled water wash 4 to 8 times, lyophilizing gets final product again.
7, the preparation method of porous tissue engineering scaffold according to claim 5 adds active particle and/or somatomedin when it is characterized in that adding porogen in step b again, and wherein active particle accounts for 5%～30% of step a copolymer gross mass; Described active particle is a kind of or wherein several mixing in gelatin, collagen, glucosan, hyaluronic acid, chitin, chitosan, alginate, hydroxyapatite, the coral reef; Somatomedin accounts for 0.5%～10% of step a copolymer gross mass; Described somatomedin is a kind of or wherein several mixing in skeletal growth factor, epidermal growth factor, insulin like growth factor, the nerve growth factor.
8, according to the preparation method of claim 5,6 or 7 described porous tissue engineering scaffolds, it is characterized in that after step b by following step at copolymer surface grafting polypeptide, concrete operations are as follows: carry out ultraviolet disinfection earlier, put into PBS solution then, add cross-linking agent again, at room temperature place 12h～48h; Putting into concentration after the taking-up is the 1g/L polypeptide solution, at room temperature places 10h～15h and promptly finishes at copolymer surface grafting polypeptide; Wherein said cross-linking agent is Sulfo-LC-SPDP, and dosage of crosslinking agent is 3%～10% of a PBS solution quality, and described polypeptide solution is by polypeptide dry powder and the preparation of PBS solution.
9, the preparation method of porous tissue engineering scaffold according to claim 8 is characterized in that described polypeptide is an arginine-glycine-aspartic acid.
10, the preparation method of porous tissue engineering scaffold, the preparation method that it is characterized in that porous tissue engineering scaffold is as follows: one, 'alpha '-hydroxy acids is carried out fusion polymerisation in bulk 24h under 110～130 ℃, the catalysis of stannous octoate, use methanol extraction after being dissolved in chloroform then, oven dry obtains polymer under 35 ℃～45 ℃ conditions again; Two, the polymer that then step 1 is obtained, dicyclohexylcarbodiimide and N-hydroxyl-butanimide activate 1～9h by 1: 5: 5 mixed in molar ratio to polymer surfaces; Three, the aminoacid cyclisation is obtained polyamino acid, the polymer of polyamino acid and surface active is added in the dimethyl sulfoxine, the total polymer mass of polyamino acid and surface active and dimethyl sulfoxine mass ratio are 1～2: 6～5, place in the deionized water behind room temperature concussion 2～5h, the mixing after-filtration, lyophilizing obtains copolymer; Four, copolymer is dissolved in to be made into mass concentration in chloroform or the ether be 5%～30% solution, add porogen again, water after sonic oscillation is extremely even and cast from the mold cavity, place 24～48h in the room temperature, dry 24～48h under 35～45 ℃ of conditions then, obtaining porosity is that 92.5%～98.5% (volume), aperture are the porous tissue engineering scaffold of 200～250 μ m; Wherein the described 'alpha '-hydroxy acids of step 3 is lactic acid and/or hydroxyacetic acid, and aminoacid accounts for 2%～40% of copolymer gross mass; Porogen is sodium chloride or sucrose in the step 4, and the mass ratio of porogen and copolymer is 4～9: 1.
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