Effects of Addition of Nano-hydroxyapatite to Highly-pressed Collagen on Osteogenic Differentiation in Osteoblastic SaOS-2 Cells
Effects of Addition of Nano-hydroxyapatite to Highly-pressed Collagen on Osteogenic Differentiation in Osteoblastic SaOS-2 Cells
复制标题
高压胶原蛋白中添加纳米羟基磷灰石对成骨细胞 SaOS-2 成骨分化的影响
DOI：
10.11344/nano.8.91
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发表时间：
2016
期刊：
影响因子：
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通讯作者：
H. Kondo
中科院分区：
文献类型：
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作者：
K. Ikeda;M. Taira;J. Yokota;M. Hattori;A. Ishisaki;H. Kondo
我们通过牛顿压机生产了高度压制的纳米羟基磷灰石/胶原复合材料（P-nHAP/COL）。本研究的目的是研究在 P-nHAP/COL 和压制胶原 (P-COL) 上培养的成骨细胞 SaOS-2 细胞的成骨分化特征，作为对照，培养 1、2、3 和 4 周。从培养细胞中提取总RNA，并使用碱性磷酸酶（ALP）、1型胶原（COL1）、骨唾液酸蛋白（BSP）和骨钙素前体（BGLAP）基因以及β-肌动蛋白基因的引物组通过实时定量（RT）PCR评估成骨分化相关基因的表达。还用扫描电子显微镜（SEM）观察培养的细胞。 RT-PCR 分析显示，P-nHAP/COL 上的成骨分化比 P-COL 上更快，特别是通过 BGLAP 基因随时间的上调而揭示。在 SEM 图像中，P-COL 上的 SaOS-2 细胞在长达 4 周内呈立方体或成纤维细胞状，但 P-nHAP/COL 上的 SaOS-2 细胞表现为颗粒状或高度拉伸。所得结果表明P-nHAP/COL可作为一种新型骨传导骨替代材料。
We produced highly pressed nano-hydroxyapatite/collagen composites (P-nHAP/COL) by Newton press. The aim of this study was to investigate osteogenic differentiation profiles in osteoblastic SaOS-2 cells cultured on P-nHAP/COL and pressed collagen (P-COL) as control for 1, 2, 3 and 4 weeks. Total RNA were extracted from cultured cells, and expressions of osteogenic-differentiation related genes were evaluated with quantitative real-time (RT) PCR using primer sets of alkaline phosphatase (ALP), type 1 collagen (COL1), bone Sialo protein (BSP) and osteocalcin precursor (BGLAP) genes as well as β-actin gene. The cultured cells were also observed with scanning electron microscopy (SEM). RT-PCR analyses revealed that osteogenic differentiation was more accelerated on P-nHAP/COL than on P-COL, especially revealed by up-regulation with time of BGLAP gene. In SEM images, SaOS-2 cells on P-COL were either cubic or fibroblastic up to 4 weeks, but those on P-nHAP/COL appeared as granular or highly stretched. The obtained results suggested that P-nHAP/COL could be employed as a new osteo-conductive bone substitute material.