Sun Junying, Wei Li, Liu Xuanyong, Li Jianyou, Li Baoe, Wang Guocheng, Meng Fanhao
Orthopedic Department, The First Affiliated Hospital of Soochow University, 188 Shizi Street, Suzhou, Jiangsu 215006, People's Republic of China.
Acta Biomater. 2009 May;5(4):1284-93. doi: 10.1016/j.actbio.2008.10.011. Epub 2008 Nov 5.
This work aims to explore the influence of the ionic products of dicalcium silicate coating on osteoblastic proliferation and differentiation, as well as on the expression of BMP2 and its signal transducers Smad1, 6 and 7 in MG-63 osteoblast-like cells. Plasma-sprayed dicalcium silicate coatings were soaked in DMEM to obtain culture media containing the ionic dissolution products of dicalcium silicate coating (Ca2SiO4-DMEM). MG63 osteoblast-like cells were cultured in Ca2SiO4-DMEM (experimental group) for 3-12 days, while those cultured in normal DMEM served as control (control group). MTT assay was used to evaluate cell viability and proliferation. Alkaline phosphatase activity (ALP), osteocalcin (OC) and type I collagen (COLI) were investigated as differentiation markers. Gene expression of BMP2 and Smad1, 6, 7 was also detected. BMP2 protein was examined by ELISA assay. Alizarin Red-S (AR-S) assay was used to detect mineralization. The results demonstrated that Si concentration in Ca2SiO4-DMEM is markedly higher than that in normal DMEM. Compared to the control group, MG63 cells of the experimental group exhibited upregulated proliferation on day 3, and markedly upregulated gene expression of the differentiation markers, especially on days 9 and 12 for OC and on days 3, 6 and 9 for ALP. Gene expression of BMP2 and Smad1, as well as BMP2 protein secreted into culture media, was also upregulated in the experimental group, while gene expression of Smad6 and 7 was not influenced. AR-S assay indicated a higher calcium mineral content deposition in cells of the experimental group. In conclusion, the ionic products of plasma-sprayed dicalcium silicate coating are beneficial to the proliferation and differentiation of MG63 osteoblast-like cells.
本研究旨在探讨硅酸二钙涂层的离子产物对成骨细胞增殖与分化的影响，以及对MG-63成骨样细胞中骨形态发生蛋白2（BMP2）及其信号转导分子Smad1、6和7表达的影响。将等离子喷涂的硅酸二钙涂层浸泡于DMEM中，以获得含有硅酸二钙涂层离子溶解产物的培养基（Ca2SiO4-DMEM）。将MG63成骨样细胞在Ca2SiO4-DMEM中培养3至12天（实验组），而在正常DMEM中培养的细胞作为对照（对照组）。采用MTT法评估细胞活力和增殖情况。检测碱性磷酸酶活性（ALP）、骨钙素（OC）和I型胶原蛋白（COLI）作为分化标志物。同时检测BMP2和Smad1、6、7的基因表达。通过ELISA法检测BMP2蛋白。采用茜素红S（AR-S）法检测矿化情况。结果表明，Ca2SiO4-DMEM中的硅浓度显著高于正常DMEM。与对照组相比，实验组的MG63细胞在第3天增殖上调，分化标志物的基因表达明显上调，尤其是OC在第9天和第12天，ALP在第3天、第6天和第9天。实验组中BMP2和Smad1的基因表达以及分泌到培养基中的BMP2蛋白也上调，而Smad6和7的基因表达未受影响。AR-S法显示实验组细胞中钙矿物质含量沉积更高。总之，等离子喷涂硅酸二钙涂层的离子产物有利于MG63成骨样细胞的增殖与分化。
