Effects of low crystalline carbonate apatite on proliferation and osteoblastic differentiation of human bone marrow cells
Effects of low crystalline carbonate apatite on proliferation and osteoblastic differentiation of human bone marrow cells
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DOI：
10.1007/s10856-015-5431-5
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发表时间：
2015-02-01
影响因子：
3.7
通讯作者：
Miyamoto, Youji
中科院分区：
文献类型：
--
作者：
Nagai, Hirokazu;Kobayashi-Fujioka, Masako;Miyamoto, Youji
碳化磷灰石(CO(3)AP)是骨的无机成分。我们提出了一种利用合成前驱体的溶解-沉淀法制备CO(3)AP块的新方法。本研究的目的是以烧结型羟基磷灰石和组织培养板为对照，研究低结晶CO(3)AP对人骨髓细胞(HBMCs)初始贴壁、增殖和成骨分化的影响。用四甲基偶氮唑盐比色法检测细胞的初始贴壁和增殖情况。逆转录-聚合酶链式反应检测成骨细胞标志物的表达。X-射线衍射仪和傅立叶变换红外光谱分析结果表明，形成了结晶度较低的B型碳酸盐磷灰石。HAP和CO(3)AP盘的初始细胞贴壁率无明显差异。HBMSC在组织培养板上的贴壁率明显高于HAP和CO(3)AP贴壁板。至第7天，HAP上的细胞数高于CO(3)AP上的细胞数，此后细胞数无明显变化。HBMSC在组织培养板上的增殖明显高于在HAP和CO(3)AP平板上的增殖。而在CO(3)AP上培养的hBMCs，其成骨分化标志物I型胶原、碱性磷酸酶、骨桥蛋白和骨钙素的表达明显高于HAP上的hBMCs。在组织培养板上，它们在整个培养过程中没有任何变化。这些结果表明，低结晶CO(3)AP具有比HAP更高的成骨活性。
Carbonated apatite (CO(3)Ap) is the inorganic component of bone. We have proposed a new method for the fabrication of CO(3)Ap blocks based on a dissolution-precipitation method using a synthetic precursor. The aim of this study is to examine the effects of low crystalline CO(3)Ap on initial cell attachment, proliferation and osteoblastic differentiation of human bone marrow cells (hBMCs) using sintered hydroxyapatite and tissue culture plates as controls. Initial cell attachment and proliferation were assessed with a MTT assay. Expression of osteoblastic markers was examined by reverse transcription-polymerase chain reaction. XRD and FT-IR results showed formation of B-type carbonate apatite with lower crystallinity. No difference was observed for initial cell attachment between HAp and CO(3)Ap discs. hBMSC attached more significantly on tissue culture plate than on HAp and CO(3)Ap discs. The number of cells on HAp was higher than that on CO(3)Ap until day 7, after which the number of cells was similar. hBMSC proliferated more significantly on tissue culture plate than on HAp and CO(3)Ap discs. In contrast, hBMCs incubated on CO(3)Ap demonstrated much higher expression of osteoblastic markers of differentiation, such as type I collagen, alkaline phosphatase, osteopontin and osteocalcin, than hBMCs on HAp. On the tissue culture plate, they were not any change throughout the culture period. These results demonstrated that low crystalline CO(3)Ap exhibit higher osteoinductivity than HAp.