不同工艺对以脐带沃顿胶制作的新型生物软骨支架的结构及理化性能的影响

发布时间：2018-01-29 04:17

本文关键词： 脐带沃顿胶软骨组织工程去细胞冷冻干燥成型软骨支架　出处：《中南大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的探索以人脐带沃顿胶制备新型生物软骨支架的方法,及其作为软骨组织工程支架载体的可行性。对不同工艺条件下所制备的支架进行大体外形和显微结构观察,并对其理化性能进行检测,研究何种工艺条件下制备的支架最符合软骨支架的理化性能要求,为新型生物软骨支架的研究提供实验依据。方法从新鲜胎儿脐带中分离出沃顿胶组织,分6组(A1、A2、A3、B1、B2、B3)经酶消化法联合物理冻融法脱细胞,再经冷冻干燥成型法制作软骨支架。其中,各组的消化时间和预冻温度依次为：30min、-196℃(A1组),30min、-80℃(A2组),30min、-20℃(A3组),1h、-196℃(B1组),1h、-80℃(B2组),1h、-20℃(B3组)。并行支架大体外形、HE染色及SEM照片观察,并对支架的孔径、孔隙率、密度、溶胀率、降解率等进行测定。结果得到的支架大体观察为表面多孔隙的白色疏松海绵泡沫样物质,在HE染色及SEM照片下观察呈多孔网状交织的三维结构,孔隙分布较均匀,材料内空隙相互连通。各组支架均脱细胞完全,经30min消化的各组消化良好,而经1h消化的各组出现消化过度现象,孔壁完整性及连续性欠佳。随预冻温度的升高,各组支架孔径、孔隙率、吸水率及降解率变大,密度减小。结论人脐带沃顿胶可制备出呈三维多孔网状结构、孔隙相互连通的生物软骨支架。其中,经30min消化、-80℃预冻制备的沃顿胶生物软骨支架去细胞完全,孔壁完整性满意,有效孔径、孔隙率、密度、吸水率及降解率等均附合软骨组织工程支架的要求。
[Abstract]:Objective to explore the method of preparing a novel biological cartilage scaffold with human umbilical cord Walton glue system, and its feasibility as a scaffold for cartilage tissue engineering carrier. To observe the general shape and microstructure of the prepared scaffolds in different conditions, and the physicochemical properties were detected, research what process conditions for the preparation of the bracket to meet the requirements of physical and chemical properties of cartilage scaffold, to provide the experimental basis for the research of novel biological cartilage scaffold.
Methods isolated from fresh umbilical cord tissue adhesive Walton, divided into 6 groups (A1, A2, A3, B1, B2, B3) by enzyme digestion combined with physical freeze thawing method of acellular cartilage scaffold, and then by making freeze drying method. Among them, each of the digestion time and freezing temperature are as follows: 30min C, -196 (group A1), 30min, -80 C (A2 group), 30min, -20 C (A3 group), 1H, -196 C (B1 group), 1H, -80 C (B2 group), 1H, -20 C (B3 group). The parallel support general shape observed by HE staining and SEM photos. And, to the support of the aperture, porosity, density, swelling ratio, degradation rate were determined.
The results obtained support general observation for white loose sponge foam like material surface of porous, three-dimensional porous structure was observed in HE staining and reticular SEM photos, the pore distribution is more homogeneous and the material pores connected with each other. All stents were decellularized completely by 30min digestion were good digestion, and after each 1H the digestive digestive excessive phenomenon, hole wall integrity and continuity is poor. With the pre freezing temperature, each scaffold pore size, porosity, water absorption rate and degradation rate becomes larger, the density decreases.
Conclusion human umbilical cord Walton glue can be prepared with a three-dimensional porous structure, biological cartilage scaffold pores connected with each other. Among them, by 30min digestion, -80 C freezing Walton glue biological cartilage scaffold prepared to complete cell wall integrity, hole, effective aperture, porosity, density, water absorption rate and degradation rate etc. all of the scaffold for cartilage tissue engineering requirements.

【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R318.08

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