CN102488926B - Tissue engineering scaffold for urethra reconstruction and preparation method thereof - Google Patents
Tissue engineering scaffold for urethra reconstruction and preparation method thereof Download PDFInfo
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Abstract
本发明涉及一种用于尿道重建的组织工程支架及其制备方法，包括：(1)制备再生丝素蛋白水溶液；(2)配制血管内皮生长因子和牛血清蛋白水溶液；(3)将(1)和(2)所制得的溶液混合，制备含血管内皮生长因子的再生丝素蛋白溶液；(4)以(3)制备的溶液为纺丝液，脱细胞基质为基层，采用静电纺丝技术制备组织工程支架。所述的组织工程支架是由脱细胞基质基层和功能层组成的双层结构工程支架，功能层为混合有血管内皮生长因子和牛血清蛋白的再生丝素蛋白纤维构成的具有三维交错网络结构特征的纤维毡。该方法制备的组织工程支架不仅具有优异的力学性能，而且能在体内早期快速血管化。本发明制备的组织工程支架用于长段尿道狭窄的修复。The invention relates to a tissue engineering scaffold for urethral reconstruction and a preparation method thereof, comprising: (1) preparing an aqueous solution of regenerated silk fibroin; (2) preparing an aqueous solution of vascular endothelial growth factor and bovine serum albumin; (3) mixing (1) Mix with the solution prepared in (2) to prepare a regenerated silk fibroin solution containing vascular endothelial growth factor; (4) use the solution prepared in (3) as the spinning solution, and the acellular matrix as the base layer, and use electrospinning technology Preparation of tissue engineering scaffolds. The tissue engineering scaffold is a double-layer structural engineering scaffold composed of an acellular matrix base layer and a functional layer. The functional layer is composed of regenerated silk fibroin fibers mixed with vascular endothelial growth factor and bovine serum albumin, and has the characteristics of a three-dimensional interlaced network structure. Fiber felt. The tissue engineering scaffold prepared by this method not only has excellent mechanical properties, but also can be rapidly vascularized early in vivo. The tissue engineering scaffold prepared by the invention is used for the repair of long section urethral stricture.
Description
Technical field
The invention belongs to the technical field of textile industry, material science, be particularly related to a kind of tissue engineering bracket for urethra reconstruction and preparation method thereof, take the regenerated silk fibroin solution that contains VEGF as spinning liquid, by electrostatic spinning process it is sprayed on the acellular matrix, has prepared the tissue engineering bracket that is used for urethra reconstruction.
Background technology
A long section complex urethral stricture not only can cause urinary extravasation, urine fistula, via etc. Local Complications, severe patient even involve other internal organs of urinary system and cause the general pathological changes has a strong impact on Quality of Life.Previously the alternative medicine take multiple autologous tissue as reconstruction biomaterials has shown good application prospect in the treatment of complex urethral stricture, but there is the shortcoming take " the sacrifice normal structure is as cost; damaged with the operation wound repair tissue " in this therapy, inevitably causes the complication at the position of drawing materials.The rise of organizational project and fast development are that new treatment approach has been opened up in the reparation of urethra pathological changes.Owing to acellular matrix (BAMG) under acellular matrix (SIS) under the small intestinal mucosa and the bladder mucosa has non-immunogenicity, antimicrobial acivity etc., can obtain metabolism by biodegradation in vivo, thereby obtain broad research and be applied clinically, but the complication such as urethra restenosis, urethral diverticulum and urine fistula easily occur in postoperative patient.Tracing it to its cause, is because of in vivo fast vascularization of acellular matrix under acellular matrix under the simple small intestinal mucosa or the bladder mucosa.Therefore promote under the small intestinal mucosa under acellular matrix or the bladder mucosa acellular matrix in vivo early stage fast vascularization be to improve the key element of its clinical application effect.
VEGF (VEGF) is a kind of effective angiogenic growth factor, can directly act on vascular endothelial cell and promote vascular endothelial cell proliferation, suppress endothelial cell apoptosis, increase vascular permeability, induce new vessels to form, but its short half-life, only exogenous VEGF added under the small intestinal mucosa under acellular matrix or the bladder mucosa can not effectively and enduringly urge its vascularization in the acellular matrix.
Solution to the problems described above: the one, adopt technique for gene engineering, make seed cell carry the VEGF gene, but the method poor controllability; The 2nd, adopt timbering material that VEGF is wrapped up, make its slow release VEGF.The key of second method is to select suitable clad material.At present the degradable clad material of bibliographical information mainly contains two kinds: the one, to be made by animal collagen or collagen fiber, and another kind is to be made by polylactic acid and medical plasticizer or collagen fiber.But after these degradation materials implant, can cause the implantation region reaction that is inflamed thereby in degradation process, easily produce acid, exist simultaneously the risk of the interior rejection of body.
And the object of the invention is to overcome above-mentioned shortcoming, select fibroin albumen as clad material.Fibroin albumen not only has excellent mechanical property, and has good biocompatibility, degradability, good breathable moisture permeability, good vascularization promoting ability etc.It is a kind of take natural silk as raw material that publication number is that the Chinese invention patent " a kind of fibre for tissue engineering scaffold and preparation method thereof " of CN 13720203A discloses, through coming unstuck, after the operation such as dissolving, adopting wet spinning technology to prepare the method for tissue engineering bracket material.The Chinese invention patent " a kind of cell culturing rack material and preparation method thereof " of publication number CN 1887362A has been announced a kind of take silkworm silk and collagen protein as raw material, adopts the Static Spinning technology to prepare the method for three-dimensional netted fibroin albumen composite nano fiber non-woven mat.But to so far, yet there are no both at home and abroad the correlational study of the compound preparation tissue engineering bracket of acellular matrix and regenerated silk fibroin under acellular matrix under the small intestinal mucosa or the bladder mucosa and bibliographical information or the patent of production.
Summary of the invention
Technical problem to be solved by this invention provides a kind of preparation method of the tissue engineering bracket for urethra reconstruction, and being the regenerated silk fibroin solution that will contain VEGF is sprayed on method on the acellular matrix by electrostatic spinning technique.The double-decker engineering rack that the tissue engineering bracket that is used for urethra reconstruction that makes by the method is comprised of basic unit and functional layer, wherein basic unit is acellular matrix, functional layer is the regenerated silk fibroin fiber felt, this fiber felt is the fiber felt with three-dimensional staggered network structure feature that is mixed with the regenerated silk fibroin fiber formation of VEGF and bovine serum albumin, VEGF is uniformly dispersed in the regenerated silk fibroin fiber, guaranteed that it can discharge and produce angiogenesispromoting effect gradually, acellular matrix is then given the mechanical property of support excellence, simultaneously, this preparation process is simple, easy operating, environmentally safe.
The object of the present invention is to provide that a kind of good biocompatibility, biological degradability are excellent, mechanical property is excellent, simultaneously can be in vivo quick tissue engineering bracket of vascularization and preparation method thereof.
A kind of double-decker engineering rack that is formed by basic unit and functional layer for the tissue engineering bracket of urethra reconstruction of the present invention; The quality of wherein said VEGF accounts for 3.5 * 10 of fiber felt gross mass
-5～9.5 * 10
-5%, the quality of described bovine serum albumin accounts for 1.5 * 10 of fiber felt gross mass
-3～4 * 10
-3%.
As preferred technical scheme:
Aforesaid a kind of tissue engineering bracket for urethra reconstruction, described acellular matrix are a kind of in the acellular matrix under acellular matrix or the bladder mucosa under the small intestinal mucosa.
Aforesaid a kind of tissue engineering bracket for urethra reconstruction, described acellular matrix are translucent cellular thin film.
Aforesaid a kind of tissue engineering bracket for urethra reconstruction, the diameter range of described regenerated silk fibroin fiber are 200nm～5 μ m.
Aforesaid a kind of tissue engineering bracket for urethra reconstruction, the porosity of described fiber felt is 50～90%.
The present invention also provides a kind of preparation method of the tissue engineering bracket for urethra reconstruction, the steps include:
(1) take Bombyx bombycis as raw material, after coming unstuck, dissolve, dialyse, concentrating, preparation quality percentage ratio is 30～40% regenerated silk fibroin water solution;
The preparation process of described regenerated silk fibroin water solution is as follows: be after 0.3～0.7% aqueous sodium carbonate comes unstuck with the Bombyx bombycis mass percent, be dissolved in the lithium bromide water solution that molar concentration is 9.0～9.5mol/L, obtain regenerated silk fibroin-lithium bromide water solution; With this solution centrifugal, filtration, dialysis and concentrated, obtain mass percent and be 30～40% regenerated silk fibroin water solution;
(2) take water as solvent, preparation VEGF and bovine serum albumin aqueous solution, the mass concentration of its VEGF is 15～25 μ g/ml, the mass concentration of bovine serum albumin is 0.6～1mg/ml;
(3) described regenerated silk fibroin water solution is mixed with 60: 1～130: 1 volume ratio with described VEGF and bovine serum albumin aqueous solution, preparation contains the regenerated silk fibroin solution of VEGF;
(4) take the regenerated silk fibroin solution that contains VEGF of step (3) preparation as spinning liquid, adopt electrostatic spinning technique that it is sprayed on the acellular matrix, for the preparation of the tissue engineering bracket of urethra reconstruction;
The preparation method of aforesaid a kind of tissue engineering bracket for urethra reconstruction, described Bombyx bombycis are a kind of in mulberry cocoon, tussah cocoon or the Semen Ricini Bombyx bombycis.
The preparation method of aforesaid a kind of tissue engineering bracket for urethra reconstruction, the described electrostatic spinning technique spinning ambient temperature during for the preparation of the tissue engineering bracket of urethra reconstruction is 2～6 ℃.
The preparation method of aforesaid a kind of tissue engineering bracket for urethra reconstruction, described acellular matrix are acellular matrix under acellular matrix or the bladder mucosa under the small intestinal mucosa.
Further, described tissue engineering bracket for urethra reconstruction is carried out post processing; Described post processing refers to that the tissue engineering bracket that will be used for urethra reconstruction soaks 30～90min at the alcohol-water mixed solution.
The preparation method of aforesaid a kind of tissue engineering bracket for urethra reconstruction, ethanol accounts for 75～95% of alcohol-water mixed solution cumulative volume in the described alcohol-water mixed solution.
Beneficial effect:
The present invention has creatively proposed to be used for the preparation method of the tissue engineering bracket of urethra reconstruction.Under the prerequisite that guarantees the regenerated silk fibroin higher concentration, avoided the use of organic solvent (such as formic acid, hexafluoroisopropanol etc.), not only reduce production cost, and avoided the injury to human body.The tissue engineering bracket that is used for urethra reconstruction that makes according to method of the present invention not only has excellent mechanical property, and in vivo fast vascularization, satisfies the instructions for use that long section urethral stricture is repaired.
The specific embodiment
Below in conjunction with the specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used for explanation the present invention and be not used in and limit the scope of the invention.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after the content of having read the present invention's instruction, these equivalent form of values fall within the application's appended claims limited range equally.
A kind of double-decker engineering rack that is formed by basic unit and functional layer for the tissue engineering bracket of urethra reconstruction of the present invention, wherein basic unit is acellular matrix, functional layer is the regenerated silk fibroin fiber felt, wherein the regenerated silk fibroin fiber felt is the fiber felt with three-dimensional staggered network structure feature that is mixed with the regenerated silk fibroin fiber formation of VEGF and bovine serum albumin, and VEGF and bovine serum albumin be Uniform Dispersion in the regenerated silk fibroin fiber; The quality of described VEGF accounts for 3.5 * 10 of described regenerated silk fibroin total fiber mass
-5～9.5 * 10
-5%, the quality of described bovine serum albumin accounts for 1.5 * 10 of described regenerated silk fibroin total fiber mass
-3～4 * 10
-3%.
Described acellular matrix is a kind of in the acellular matrix under acellular matrix or the bladder mucosa under the small intestinal mucosa.
Described acellular matrix is translucent cellular thin film.
The diameter range of the regenerated silk fibroin fiber of described formation fiber felt is 200nm～5 μ m.
The porosity of described fiber felt is 50～90%.
Tissue engineering bracket of the present invention is used for the reparation of long section urethral stricture.
Embodiment 1
A kind of preparation method of the tissue engineering bracket for urethra reconstruction the steps include:
(1) take mulberry cocoon as raw material, after coming unstuck, dissolve, dialyse, concentrating, preparation quality percentage ratio is 30% regenerated silk fibroin water solution;
The preparation process of described regeneration pure silk fibroin aqueous solution is as follows: be after 0.3% aqueous sodium carbonate comes unstuck with mulberry cocoon with mass percent, be dissolved in the lithium bromide water solution that molar concentration is 9.0mol/L, obtain regenerated silk fibroin-lithium bromide water solution; With this solution centrifugal, filtration, dialysis and concentrated, obtain mass percent and be 30% regenerated silk fibroin water solution;
(2) take water as solvent, preparation VEGF and bovine serum albumin aqueous solution, the mass concentration of its VEGF is 15 μ g/ml, the mass concentration of bovine serum albumin is 0.6mg/ml;
(3) described regenerated silk fibroin water solution is mixed with 130: 1 volume ratio with described VEGF and bovine serum albumin aqueous solution, preparation contains the regenerated silk fibroin solution of VEGF;
(4) take the regenerated silk fibroin solution that contains VEGF of step (3) preparation as spinning liquid, it is sprayed under the small intestinal mucosa on the acellular matrix, for the preparation of the tissue engineering bracket of urethra reconstruction at 2 ℃ of lower electrostatic spinning techniques that adopt; The diameter range of the regenerated silk fibroin fiber of described formation tissue engineering bracket functional layer is 200nm～1 μ m, and the porosity of described fiber felt is 90%.
Embodiment 2
A kind of preparation method of the tissue engineering bracket for urethra reconstruction the steps include:
(1) take tussah cocoon as raw material, after coming unstuck, dissolve, dialyse, concentrating, preparation quality percentage ratio is 33% regenerated silk fibroin water solution;
The preparation process of described regenerated silk fibroin water solution is as follows: be after 0.4% aqueous sodium carbonate comes unstuck, to be dissolved in the lithium bromide water solution that molar concentration is 9.1mol/L with the tussah cocoon mass percent, obtain regenerated silk fibroin-lithium bromide water solution; With this solution centrifugal, filtration, dialysis and concentrated, obtain mass percent and be 33% regenerated silk fibroin water solution;
(2) take water as solvent, preparation VEGF and bovine serum albumin aqueous solution, the mass concentration of its VEGF is 17 μ g/ml, the mass concentration of bovine serum albumin is 0.7mg/ml;
(3) described regenerated silk fibroin water solution is mixed with 70: 1 volume ratio with described VEGF and bovine serum albumin aqueous solution, preparation contains the regenerated silk fibroin solution of VEGF;
(4) take the regenerated silk fibroin solution that contains VEGF of step (3) preparation as spinning liquid, it is sprayed under the bladder mucosa on the acellular matrix, for the preparation of the tissue engineering bracket of urethra reconstruction at 4 ℃ of lower electrostatic spinning techniques that adopt; The diameter range of the regenerated silk fibroin fiber of described formation tissue engineering bracket functional layer is 500nm～2 μ m, and the porosity of described fiber felt is 81%.
Embodiment 3
A kind of preparation method of the tissue engineering bracket for urethra reconstruction the steps include:
(1) take Semen Ricini Bombyx bombycis as raw material, after coming unstuck, dissolve, dialyse, concentrating, preparation quality percentage ratio is 35% regenerated silk fibroin water solution;
The preparation process of described regenerated silk fibroin water solution is as follows: be after 0.5% aqueous sodium carbonate comes unstuck with Semen Ricini Bombyx bombycis mass percent, be dissolved in the lithium bromide water solution that molar concentration is 9.2mol/L, obtain regenerated silk fibroin-lithium bromide water solution; With this solution centrifugal, filtration, dialysis and concentrated, obtain mass percent and be 35% regenerated silk fibroin water solution;
(2) take water as solvent, preparation VEGF and bovine serum albumin aqueous solution, the mass concentration of its VEGF is 20 μ g/ml, the mass concentration of bovine serum albumin is 0.8mg/ml;
(3) described regenerated silk fibroin water solution is mixed with 90: 1 volume ratio with described VEGF and bovine serum albumin aqueous solution, preparation contains the regenerated silk fibroin solution of VEGF;
(4) take the regenerated silk fibroin solution that contains VEGF of step (3) preparation as spinning liquid, it is sprayed under the bladder mucosa on the acellular matrix, for the preparation of the tissue engineering bracket of urethra reconstruction at 6 ℃ of lower electrostatic spinning techniques that adopt; The diameter range of the regenerated silk fibroin fiber of described formation tissue engineering bracket functional layer is 800nm～3 μ m, and the porosity of described fiber felt is 72%.
Embodiment 4
A kind of preparation method of the tissue engineering bracket for urethra reconstruction the steps include:
(1) take Semen Ricini Bombyx bombycis as raw material, after coming unstuck, dissolve, dialyse, concentrating, preparation quality percentage ratio is 37% regenerated silk fibroin water solution;
The preparation process of described regenerated silk fibroin water solution is as follows: be after 0.6% aqueous sodium carbonate comes unstuck with Semen Ricini Bombyx bombycis mass percent, be dissolved in the lithium bromide water solution that molar concentration is 9.3mol/L, obtain regenerated silk fibroin-lithium bromide water solution; With this solution centrifugal, filtration, dialysis and concentrated, obtain mass percent and be 37% regenerated silk fibroin water solution;
(2) take water as solvent, preparation VEGF and bovine serum albumin aqueous solution, the mass concentration of its VEGF is 22 μ g/ml, the mass concentration of bovine serum albumin is 0.9mg/ml;
(3) described regenerated silk fibroin water solution is mixed with 110: 1 volume ratio with described VEGF and bovine serum albumin aqueous solution, preparation contains the regenerated silk fibroin solution of VEGF;
(4) take the regenerated silk fibroin solution that contains VEGF of step (3) preparation as spinning liquid, it is sprayed under the small intestinal mucosa on the acellular matrix, for the preparation of the tissue engineering bracket of urethra reconstruction at 4 ℃ of lower electrostatic spinning techniques that adopt; The diameter range of the regenerated silk fibroin fiber of described formation tissue engineering bracket functional layer is 1 μ m～4 μ m, and the porosity of described fiber felt is 59%.
Embodiment 5
A kind of preparation method of the tissue engineering bracket for urethra reconstruction the steps include:
(1) take mulberry cocoon as raw material, after coming unstuck, dissolve, dialyse, concentrating, preparation quality percentage ratio is 40% regenerated silk fibroin water solution;
The preparation process of described regenerated silk fibroin water solution is as follows: be after 0.7% aqueous sodium carbonate comes unstuck, to be dissolved in the lithium bromide water solution that molar concentration is 9.5mol/L mulberry cocoon with mass percent, obtain regenerated silk fibroin-lithium bromide water solution; With this solution centrifugal, filtration, dialysis and concentrated, obtain mass percent and be 40% regenerated silk fibroin water solution;
(2) take water as solvent, preparation VEGF and bovine serum albumin aqueous solution, the mass concentration of its VEGF is 25 μ g/ml, the mass concentration of bovine serum albumin is 1.0mg/ml;
(3) described regenerated silk fibroin water solution is mixed with 60: 1 volume ratio with described VEGF and bovine serum albumin aqueous solution, preparation contains the regenerated silk fibroin solution of VEGF;
(4) take the regenerated silk fibroin solution that contains VEGF of step (3) preparation as spinning liquid, it is sprayed under the small intestinal mucosa on the acellular matrix, for the preparation of the tissue engineering bracket of urethra reconstruction at 6 ℃ of lower electrostatic spinning techniques that adopt; The diameter range of the regenerated silk fibroin fiber of described formation tissue engineering bracket functional layer is 1.5 μ m～5 μ m, and the porosity of described fiber felt is 50%.
Embodiment 6
A kind of preparation method of the tissue engineering bracket for urethra reconstruction the steps include:
(1) take mulberry cocoon as raw material, after coming unstuck, dissolve, dialyse, concentrating, preparation quality percentage ratio is 35% regenerated silk fibroin water solution;
The preparation process of described regenerated silk fibroin water solution is as follows: be after 0.7% aqueous sodium carbonate comes unstuck, to be dissolved in the lithium bromide water solution that molar concentration is 9.2mol/L mulberry cocoon with mass percent, obtain regenerated silk fibroin-lithium bromide water solution; With this solution centrifugal, filtration, dialysis and concentrated, obtain mass percent and be 35% regenerated silk fibroin water solution;
(2) take water as solvent, preparation VEGF and bovine serum albumin aqueous solution, the mass concentration of its VEGF is 22 μ g/ml, the mass concentration of bovine serum albumin is 0.9mg/ml;
(3) described regenerated silk fibroin water solution is mixed with 90: 1 volume ratio with described VEGF and bovine serum albumin aqueous solution, preparation contains the regenerated silk fibroin solution of VEGF;
(4) take the regenerated silk fibroin solution that contains VEGF of step (3) preparation as spinning liquid, it is sprayed under the bladder mucosa on the acellular matrix preparation tissue engineering bracket at 2 ℃ of lower electrostatic spinning techniques that adopt; The diameter range of the regenerated silk fibroin fiber of described formation tissue engineering bracket functional layer is 800nm～3 μ m, and the porosity of described fiber felt is 72%;
(5) tissue engineering bracket of step (4) preparation is soaked 90min in volume fraction of ethanol is 75% alcohol-water mixed solution, namely obtain finally being used for the tissue engineering bracket of urethra reconstruction.
Embodiment 7
A kind of preparation method of the tissue engineering bracket for urethra reconstruction the steps include:
(1) take tussah cocoon as raw material, after coming unstuck, dissolve, dialyse, concentrating, preparation quality percentage ratio is 37% regenerated silk fibroin water solution;
The preparation process of described regenerated silk fibroin water solution is as follows: be after 0.7% aqueous sodium carbonate comes unstuck, to be dissolved in the lithium bromide water solution that molar concentration is 9.0mol/L with the tussah cocoon mass percent, obtain regenerated silk fibroin-lithium bromide water solution; With this solution centrifugal, filtration, dialysis and concentrated, obtain mass percent and be 37% regenerated silk fibroin water solution;
(2) take water as solvent, preparation VEGF and bovine serum albumin aqueous solution, the mass concentration of its VEGF is 17 μ g/ml, the mass concentration of bovine serum albumin is 0.7mg/ml;
(3) described regenerated silk fibroin water solution is mixed with 110: 1 volume ratio with described VEGF and bovine serum albumin aqueous solution, preparation contains the regenerated silk fibroin solution of VEGF;
(4) take the regenerated silk fibroin solution that contains VEGF of step (3) preparation as spinning liquid, it is sprayed under the small intestinal mucosa on the acellular matrix preparation tissue engineering bracket at 4 ℃ of lower electrostatic spinning techniques that adopt; The diameter range of the regenerated silk fibroin fiber of described formation tissue engineering bracket functional layer is 1 μ m～4 μ m, and the porosity of described fiber felt is 61%;
(5) tissue engineering bracket of step (4) preparation is soaked 60min in volume fraction of ethanol is 85% alcohol-water mixed solution, namely obtain finally being used for the tissue engineering bracket of urethra reconstruction.
Embodiment 8
A kind of preparation method of the tissue engineering bracket for urethra reconstruction the steps include:
(1) take tussah cocoon as raw material, after coming unstuck, dissolve, dialyse, concentrating, preparation quality percentage ratio is 33% regenerated silk fibroin water solution;
The preparation process of described regenerated silk fibroin water solution is as follows: be after 0.5% aqueous sodium carbonate comes unstuck, to be dissolved in the lithium bromide water solution that molar concentration is 9.1mol/L with the tussah cocoon mass percent, obtain regenerated silk fibroin-lithium bromide water solution; With this solution centrifugal, filtration, dialysis and concentrated, obtain mass percent and be 33% regenerated silk fibroin water solution;
(2) take water as solvent, preparation VEGF and bovine serum albumin aqueous solution, the mass concentration of its VEGF is 19 μ g/ml, the mass concentration of bovine serum albumin is 0.8mg/ml;
(3) described regenerated silk fibroin water solution is mixed with 70: 1 volume ratio with described VEGF and bovine serum albumin aqueous solution, preparation contains the regenerated silk fibroin solution of VEGF;
(4) take the regenerated silk fibroin solution that contains VEGF of step (3) preparation as spinning liquid, it is sprayed under the small intestinal mucosa on the acellular matrix preparation tissue engineering bracket at 6 ℃ of lower electrostatic spinning techniques that adopt; The diameter range of the regenerated silk fibroin fiber of described formation tissue engineering bracket functional layer is 500nm～2 μ m, and the porosity of described fiber felt is 81%;
(5) tissue engineering bracket of step (4) preparation is soaked 30min in volume fraction of ethanol is 95% alcohol-water mixed solution, namely obtain finally being used for the tissue engineering bracket of urethra reconstruction.
Claims (10)
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| CN102813965B (en) * | 2012-08-09 | 2015-04-01 | 东华大学 | Multilayer well-organized silk fibroin tissue engineering scaffold and preparation method thereof |
| CN103480041B (en) * | 2013-09-05 | 2015-05-27 | 陕西瑞盛生物科技有限公司 | Tendon reinforcing repairing material and preparation method thereof |
| CN104473706B (en) * | 2014-12-09 | 2017-02-22 | 金华市人民医院 | Biodegradable composite type tubular urethral stent and preparation method |
| CN105505883A (en) * | 2016-01-12 | 2016-04-20 | 河南师范大学 | Optimized culture method for primary rat hepatocytes |
| CN106075586B (en) * | 2016-07-19 | 2019-05-24 | 东华大学 | Cultural method based on adult stem cell-regenerated silk fibroin tissue engineering bracket artificial liver repair materials |
| CN107551322A (en) * | 2017-10-31 | 2018-01-09 | 无锡中科光远生物材料有限公司 | A kind of compound support frame material for Bladder repair |
| CN110538348A (en) * | 2018-06-05 | 2019-12-06 | 广州市妇女儿童医疗中心 | A kind of urethral tissue engineering scaffold and preparation method thereof |
| CN109248341B (en) * | 2018-11-20 | 2020-10-09 | 北京航空航天大学 | Soft tissue repair patch and preparation method thereof |
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| CN109847101B (en) * | 2018-12-28 | 2022-04-12 | 广州市妇女儿童医疗中心 | A kind of tissue engineering urethral stent and preparation process thereof |
| CN113577401B (en) * | 2021-07-15 | 2022-06-07 | 南通纺织丝绸产业技术研究院 | A long-acting antibacterial and anti-stenosis functional urethral stent and preparation method thereof |
| CN114949322A (en) * | 2022-04-13 | 2022-08-30 | 齐齐哈尔医学院 | Preparation method of tissue-induced nanofiber biomaterial capable of trapping host cells by graphene quantum dot tweezers and homing |
| CN115414533B (en) * | 2022-09-20 | 2024-02-23 | 中国人民解放军总医院第一医学中心 | Tissue engineering scaffold simulating physiological structure of bladder wall and preparation method thereof |
| CN115487359B (en) * | 2022-09-26 | 2023-06-02 | 中国人民解放军总医院第一医学中心 | Angiogenesis promoting bladder tissue engineering patch, preparation method and application thereof |
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| US8252738B2 (en) * | 1999-04-30 | 2012-08-28 | Warsaw Orthopedic, Inc. | Method for changing cell phenotype using LIM mineralization proteins |
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| US9102916B2 (en) * | 2007-02-27 | 2015-08-11 | Trustees Of Tufts College | Tissue-engineered silk organs |
| CN100548391C (en) * | 2007-11-09 | 2009-10-14 | 北京理工大学 | The preparation method of fibroin albumen antipriming pipe |
| CN101444645A (en) * | 2007-11-28 | 2009-06-03 | 上海交通大学医学院附属第九人民医院 | Method for preparing mucosal graft |
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