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目的:柞蚕丝素(tussah silk fibroin,TSF)和羟基磷灰石(hydroxyapatite,HA)均具有良好的生物活性和生物相容性,是组织工程研究的热点。但结构单一及微米级的材料所表现出的性能简单,不能满足人们对生物材料支架性能的要求,本课题将两者按不同比例进行复合,探讨不同皮芯比例羟基磷灰石/柞蚕丝素(HA/TSF)的骨仿生纳米纤维的生物学性能。方法:首先利用同轴静电纺丝技术,以TSF水溶液为皮,HA水溶液为芯,制备不同皮芯比例的HA/TSF骨仿生纳米纤维,然后将人成骨肉瘤细胞(MG-63)种植在不同皮芯比例的HA/TSF纳米纤维上。在不同的时间点分别通过倒置显微镜、扫描电镜观察细胞形态学变化;通过四甲基偶氮噻唑蓝比色(Four methyl azo thiazole blue colorimetric,MTT)法、碱性磷酸酶(alkaline phosphatase,ALP)活性检测法观察细胞在材料表面的增殖和分化,从多角度来评价材料的生物学性能。结果:通过形态学观察,SEM观察以及MTT检测,发现除空白对照组外,各组样品均显示良好的生物相容性,均能促进细胞的黏附、增殖,尤以HA/TSF为2:1时最明显;通过MG-63细胞的ALP活性检测,发现当HA/TSF比例为2:1时,最能促进细胞ALP活性的表达,有利于诱导成骨细胞的分化。结论:皮芯结构的HA/TSF骨仿生纳米纤维具有良好的生物学性能,且二者在自然界来源丰富,价格便宜,为临床骨组织缺损修复的应用奠定了一定的实验基础。
OBJECTIVE: Tussah silk fibroin (TSF) and hydroxyapatite (HA) have good biological activity and biocompatibility, which are the hot topics in tissue engineering. However, the properties of single and micron-sized materials are simple and can not meet people’s requirements for the performance of biomaterial scaffolds. In this project, the two materials are compounded in different proportions to investigate the effects of different ratios of hydroxyapatite / tussah silk fibroin (HA / TSF) bone biomimetic nanofibers biological properties. METHODS: HA / TSF bone biomimetic nanofibers with different core / sheath ratios were prepared by coaxial electrospinning using TSF aqueous solution and HA aqueous solution as the core. Then, human osteosarcoma cells (MG-63) were seeded on Different sheath / core ratios of HA / TSF nanofibers. The morphological changes of the cells were observed by inverted microscope and scanning electron microscope at different time points. The cells were stained with MTT (alkaline phosphatase), alkaline phosphatase (ALP) Activity assay was used to observe the proliferation and differentiation of cells on the surface of the material, and to evaluate the biological properties of the material from multiple perspectives. Results: Morphological observation, SEM observation and MTT assay showed that all the samples showed good biocompatibility except for the blank control group, which could promote the cell adhesion and proliferation, especially the HA / TSF was 2: 1 When the ratio of HA / TSF was 2: 1, it was found that ALP activity of MG-63 cells could promote the differentiation of osteoblasts. CONCLUSION: HA / TSF bone biomimetic nanofibers with sheath-core structure have good biological properties. Both of them are abundant in nature and cheap in price, which has laid a solid experimental foundation for the clinical application of bone tissue repair.