N-乙酰半胱氨酸丝素微球复合磷酸钙骨水泥的制备及其性能的研究

发布时间：2018-02-03 04:33

本文关键词： 生物材料 N-乙酰半胱氨酸丝素蛋白微球磷酸钙骨水泥药物缓释细胞增殖　出处：《苏州大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的:制备复合N-乙酰半胱氨酸丝素微球可注射性磷酸钙骨水泥,构建一种具有药物缓释功能的新型注射性植骨材料,并评价其理化性能及细胞相容性。方法:采用乳化溶剂蒸发法制备空丝素微球,并用粒度仪测定微球粒径,扫描电镜观察微球表面形态;用空丝素微球吸附不同浓度N-乙酰半胱氨酸溶液,确定最大理论载药率时N-乙酰半胱氨酸浓度,再将载有N-乙酰半胱氨酸的丝素微球载入磷酸钙骨水泥中,检测负载N-乙酰半胱氨酸丝素微球的磷酸钙骨水泥的体外释药性能;体外将小鼠MC3T3-E1成骨细胞与N-乙酰半胱氨酸丝素微球磷酸钙骨水泥共培养,扫描电镜观察细胞在材料表面的贴附及生长。将单纯磷酸钙、丝素微球磷酸钙、N-乙酰半胱氨酸丝素微球磷酸钙骨水泥共3种骨水泥浸提液,以及含有体积分数10%胎牛血清和1%青霉素-链霉素的α-MEM培养基,与MC3T3-E1细胞共培养,对照组仅用上述培养基培养,采用MTS法评估各组细胞增殖情况。结果:(1)粒径分析显示丝素微球直径集中在2-10μm范围内,平均粒径为3.20μm,扫描电镜示复合骨水泥中的微球表面光滑平整,大小基本均一,散在分布,可见在制备复合骨水泥的过程中微球能保持稳定,不被破坏。(2)用丝素溶液作为固化液后磷酸钙骨水泥的初凝、终凝时间较用去离子水作为固化液明显缩短(P0.05)(3)采用丝素微球、磷酸钙骨水泥作为药物双重缓释体系在前24 h累计释药(5.35±2.19)%,较对照组(13.45±1.04)%明显减少,28 d时对照组累计释药(37.64±1.73)%,实验组累计释药(18.99±0.92)%。(4)MTS细胞增殖实验中,各组材料浸提液对MC3T3-E1细胞的生长较对照组无明显差异,扫描电镜示材料表面细胞形态呈梭型,生长情况良好。结论:1.丝素微球的加入使得磷酸钙骨水泥对药物的缓释速度得以减慢。2.丝素蛋白溶液可缩短磷酸钙骨水泥的凝固时间,提升操作性。3.复合骨水泥材料具有良好的生物相容性,有望为骨修复生物材料的研究提供参考。
[Abstract]:Objective: to prepare injectable calcium phosphate cement with N-acetylcysteine fibroin microspheres and to construct a new injectable bone graft material with drug delivery function. Methods: the empty fibroin microspheres were prepared by emulsified solvent evaporation method. The particle size was measured by particle size analyzer and the surface morphology of the microspheres was observed by scanning electron microscope (SEM). Different concentrations of N-acetylcysteine solution were adsorbed with empty fibroin microspheres, and the concentration of N-acetylcysteine was determined when the maximum theoretical drug loading rate was determined. Then the silk fibroin microspheres loaded with N-acetylcysteine were loaded into calcium phosphate cement to detect the drug release properties of calcium phosphate bone cement loaded with N-acetylcysteine fibroin microspheres. Mouse MC3T3-E1 osteoblasts were co-cultured with N-acetylcysteine fibroin microspheres calcium phosphate bone cement in vitro. The adhesion and growth of the cells on the surface of the materials were observed by scanning electron microscope. Silk fibroin microspheres calcium phosphate N-acetylcysteine fibroin microspheres calcium phosphate bone cement were extracted from three kinds of bone cement. The 伪 -MEM medium containing 10% fetal bovine serum and 1% penicillin streptomycin was co-cultured with MC3T3-E1 cells. Results the diameter of fibroin microspheres was in the range of 2-10 渭 m, and the average diameter was 3.20 渭 m. Scanning electron microscopy showed that the surface of the composite bone cement was smooth and smooth, the size of the microspheres was almost uniform, and the microspheres were scattered. It can be seen that the microspheres remained stable during the preparation of the composite bone cement. The initial setting time of calcium phosphate bone cement with silk fibroin solution as solidification solution was significantly shorter than that of deionized water as curing solution. The cumulative release of calcium phosphate cement was 5.35 卤2.19% in the first 24 hours, which was significantly lower than that in the control group (13.45 卤1.04%). On the 28th day, the cumulative drug release of the control group was 37.64 卤1.73 and that of the experimental group was 18.99 卤0.92 in the proliferation test of MTS cells. There was no significant difference in the growth of MC3T3-E1 cells in each group compared with the control group. The morphology of the cells on the surface of the materials was fusiform by scanning electron microscope. Conclusion: the addition of silk fibroin microspheres can slow down the slow release rate of calcium phosphate cement. 2. Silk fibroin solution can shorten the solidification time of calcium phosphate cement. The composite bone cement has good biocompatibility, which is expected to provide a reference for the study of bone repair biomaterials.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R318.08;R687

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