探究牙髓干细胞在壳聚糖-纤维蛋白原复合支架上的黏附与增殖

发布时间：2018-03-09 14:22

本文选题：壳聚糖　切入点：纤维蛋白原　出处：《新疆医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:检测牙髓干细胞在壳聚糖-纤维蛋白原复合支架材料上的黏附与增殖情况。方法:(1)采用改良酶消化法分离提取C57乳鼠牙髓干细胞,传代培养至第3代,体外成脂成骨诱导分化对细胞进行鉴定;(2)测定壳聚糖-纤维蛋白原复合支架材料的孔径及孔隙率;(3)将壳聚糖-纤维蛋白原复合支架材料与第3代小鼠牙髓干细胞共培养,MTT法检测细胞1~8天的增殖情况;(4)扫描电镜观察壳聚糖-纤维蛋白原复合支架材料的形态,细胞黏附、增殖及基质分泌情况。结果:(1)成功分离培养C57小鼠牙髓干细胞,经成脂成骨诱导分化后结果呈阳性;(2)壳聚糖-纤维蛋白原复合支架材料孔径为(105.32±22.10)μm,孔隙率为(87.714±1.276)%;(3)牙髓干细胞在壳聚糖-纤维蛋白原复合支架材料上的增殖曲线与对照组基本一致,均为“S”型,培养第4~8天实验组增殖速度显著快于对照组,差异有显著性意义(P0.05);(4)扫描电镜下细胞接种第2天与壳聚糖-纤维蛋白原复合支架材料黏附紧密,生长状态良好,接种第4天,细胞明显开始增殖,体积增大,有丰富的细胞外基质,接种第6天,铺满与整个材料表面及空隙。结论:(1)牙髓干细胞提取便捷,并表现出成脂成骨的生物学特性,是适合牙髓组织工程的种子细胞。(2)壳聚糖-纤维蛋白原复合材料是一种适宜牙髓干细胞黏附与增殖的支架材料。
[Abstract]:Objective: to investigate the adhesion and proliferation of dental pulp stem cells on chitosan / fibrinogen composite scaffolds. Methods: C57 dental pulp stem cells were isolated and cultured to the third generation by modified enzyme digestion method. Determination of pore size and porosity of chitosan / fibrinogen composite scaffold material by in vitro adipogenic osteogenic induction and differentiation) the chitosan / fibrinogen composite scaffold material and the third generation mouse dental pulp stem cells were used for the determination of pore size and porosity of chitosan / fibrinogen composite scaffold. MTT assay was used to detect the proliferation of cells for 8 days. Scanning electron microscope (SEM) was used to observe the morphology of chitosan fibrinogen composite scaffold. Results the dental pulp stem cells of C57 mice were isolated and cultured successfully by cell adhesion, proliferation and matrix secretion. The results of adipogenic osteogenesis induced differentiation were positive. The pore size of chitosan / fibrinogen composite scaffold was 105.32 卤22.10 渭 m, the porosity was 87.714 卤1.276%. The proliferation curve of dental pulp stem cells on chitosan fibrinogen composite scaffold was basically the same as that of control group. The proliferation rate of the experimental group was significantly faster than that of the control group on the 4th day of culture. The difference was significant (P 0.05). Under the scanning electron microscope (SEM), the cells adhered closely to the chitosan / fibrinogen composite scaffold material on the 2nd day, and the growth state was good. On the 4th day of inoculation, the cells began to proliferate obviously, the volume increased, and there was abundant extracellular matrix. On the 6th day of inoculation, the cells were covered with the whole material surface and space. Conclusion the dental pulp stem cells are easy to extract and show the biological characteristics of adipogenic osteogenesis. Chitosan-fibrinogen composite is a kind of scaffold material suitable for dental pulp stem cell adhesion and proliferation.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R783.1

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