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目的研究以硅在天然骨中的含量为掺杂量制备的硅-羟基磷灰石(Si-HA)对成骨细胞活性的影响。方法以水热法制备微量硅掺杂改性的Si-HA;通过X射线衍射(X-ray diffraction,XRD)及扫描电镜观察材料表面构相;MG63细胞在材料表面分别接种培养1、6、12h及1、4、7、14d,通过CCK8方法分别检测MG63细胞在材料表面的黏附、增殖情况;通过DAPI染色法验证CCK8检测细胞黏附数量;并对1、4、7、14d成骨细胞碱性磷酸酶(ALP)活性进行检测;采用RTPCR检测成骨细胞特异性基因ALP、Ⅰ型胶原(Col-Ⅰ)、骨钙素(OC)在1、4、7、14d的表达。结果 Si的掺杂未改变HA表面结构及组成;MG63细胞在材料表面培养6、12h,Si-HA表面细胞附着数量明显高于HA表面(P<0.05);DAPI染色法与CCK8实验结果相同;细胞培养4、7d时,Si-HA较HA表面的增殖细胞数量增多(P<0.05)。细胞培养4、7d时,Si-HA材料表面ALP活性及基因表达高于HA表面(P<0.05);4、7、14d时Col-Ⅰ基因在Si-HA表面的表达量明显高于HA表面(P<0.05)。7、14d时,OC基因在Si-HA表面的表达量明显高于HA表面(P<0.05)。结论以硅在天然骨中的含量为掺杂量的Si-HA能够促进MG63成骨细胞黏附、增殖,上调成骨特异性基因的表达,提高了HA材料的生物学活性。
Objective To study the effect of silicon-hydroxyapatite (Si-HA) prepared by the content of silicon in natural bone on the activity of osteoblasts. Methods The Si-HA modified by silicon was prepared by hydrothermal method. The surface morphology of the material was observed by X-ray diffraction (XRD) and scanning electron microscopy (SEM). MG63 cells were seeded on the surface of the material for 1,6, 12 h and 1, 4, 7 and 14 days. The adhesion and proliferation of MG63 cells on the surface of the material were detected by CCK8 assay. The number of cells adhered by CCK8 was detected by DAPI staining. The activity of ALP was detected. The expression of osteoblast-specific genes ALP, Col-Ⅰ and OC were detected by RT-PCR at 1, 4, 7 and 14 days. Results The doping of Si did not change the structure and composition of the surface of HA. MG63 cells were cultured on the surface of the material for 6 and 12 hours, and the number of cells attached to the surface of Si-HA was significantly higher than that of HA surface (P <0.05) At 4 and 7 days after culture, the number of proliferating cells in Si-HA increased compared with HA surface (P <0.05). At 4 and 7 days, the ALP activity and gene expression on Si-HA surface were higher than those on HA surface (P <0.05), and the expression of Col-Ⅰ gene on Si-HA surface was significantly higher than that on HA surface (P <0.05). At 7 and 14 days, the expression of OC gene on Si-HA surface was significantly higher than that on HA surface (P <0.05). Conclusion Si-HA, which contains silicon in natural bone, can promote the osteoblast adhesion and proliferation, up-regulate the expression of osteoblast-specific gene and increase the biological activity of HA material.