硅酸钙、β-磷酸三钙和羟基磷灰石三相陶瓷复合物与骨细胞活性的体外鉴定(英文)

In vitro characterization of bone cell activity on triphasic ceramic composites from calcium silicate,beta-tricalcium phosphate and hydroxyapatite

由于羟基磷灰石陶瓷与骨无机质的化学成份相似,所以10年来研究者对羟基磷灰石陶瓷在骨修复替换中的应用进行了深入的研究。羟基磷灰石因其高生物相容性和骨传导性已被广泛的应用,但羟基磷灰石的再吸收能力差。文章对由硅酸钙、β-磷酸三钙和羟基磷灰石组成的三相陶瓷复合物进行了研究。在三相陶瓷复合物环境下,采用不同比率硅酸钙、β-磷酸三钙和羟基磷灰石对骨髓夹源间充质干细胞进行体外研究。细胞在加入和没有加入成骨细胞的情况下培养28d。采用碱性磷酸酶活性、碱性磷酸酶基因码转录和骨唾液蛋白Ⅱ表达评估成骨分化水平。成骨细胞和破骨细胞在陶瓷复合80%羟基磷灰石材料上完成预实验。将鼠颅盖骨成骨细胞与人单核细胞共同培养以分析其向破骨细胞分化的可能性。结果显示人骨髓间充质干细胞在陶瓷复合物材料上表现出快速黏附和高度增殖率,在所有实验材料上,成骨诱导的骨髓间充质干细胞碱性磷酸酶活性增加,碱性磷酸酶基因码转录和骨唾液酸蛋白Ⅱ的表达在3种材料上同样增加。扫描电镜和聚焦激光扫描显微镜结果证明在陶瓷复合80%羟基磷灰石材料上的人单核细胞向破骨样细胞分化。结果提示制备的三相陶瓷复合材料支持人骨髓间充质干细胞的黏附、增殖和成骨分化,以及人单核细胞与成骨细胞联合培养条件下破骨细胞形成。

Hydroxyapatite（HA）ceramics have been deeply studied over the last decades for application in bone replacement.Due to its close chemical similarity to bone mineral,synthetic HA has been extensively used in clinical applications by virtue of its high biocompatibility and osteoconductivity but shows poor resorbability.We developed a triphasic ceramic composite comprising of HA,beta-tricalcium phosphate（beta-TCP）and calcium silicate.In vitro studies applying human bone marrow-derived mesenchymal stem cells（hMSC）were performed on three different types of triphasic ceramic composites,containing varying weight ratios of HA and beta-TCP/calcium silicate.Cells were cultivated over 28 days with and without osteogenic supplements.To study remodelling,a preliminary experiment for co-culture of osteoblasts and osteoclasts was performed on ceramic composite containing 80% HA.Rat calvarial osteoblasts were cultured together with human monocytes which can be differentiated into osteoclasts.Results showed that hMSC exhibited rapid adhesion and high proliferation rates on the ceramic composite materials.Osteogenic induced hMSC showed an increase of alkaline phosphatase activity on all examined materials.Transcription of genes coding for alkaline phosphatase and bone sialoprotein II was also increased for all three materials.Scanning electron microscopy and confocal laser scanning microscopy investigations demonstrated that osteoclast-like cells were developed from human monocytes on ceramics containing 80% HA.These findings indicate that the novel prepared triphasic ceramic composites support adhesion,proliferation and osteogenic differentiation of hMSC as well as osteoclastogenesis of human monocytes in co-culture with osteoblasts.