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A Trial to Establish New Methods for the Evaluation of Bio-affinity of Metal Surfaces by using Cell Culture System

利用细胞培养系统建立金属表面生物亲和力评价新方法的尝试

基本信息

批准号：
18590186
负责人：
KOBAYASHI Naoto
金额：
$ 2.52万
依托单位：
Ehime University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

The present study aimed to analyze, by using in vitro cell cultures of human osteoblasts, the dynamic cellular behavior on the clinical materials including metal specimens, and to establish new methods for the evaluation of bio-affinity of such materials. The present study has revealed following data. 1) By using a human cell line of osteoblasts (Saos-2), their behavior on the titanium surface was analyzed in the view point of cell spreading and cell motility with a special reference on the surface smoothness (Li C, Gao S, Kobayashi N, et. al., 2006). Furthermore, intracellular signals regulating cell motility were assayed by the same system to show that actin cytoskeleton has a primary role in the cell motility of Saos-2 cells and that the direction of cell motility is thought to be regulated by IP3-kinase signals. 2) By using a primary culture system of human osteoblasts, a new method has been established to evaluate cell motility on clinical materials. When cells were seeded, a glass cover slip was placed in the center of a plastic culture dish and the cover slip was pressed by a weight. After cells were attached on the dish, the cover slip was removed to obtain a cell-free rectangle region, into which cells started to migrate allowing the measurement of cell motility along the time course. After this simple method, cell motility of the primary cultured cells is shown to be analyzed with a significant reproductivity.

本研究旨在利用体外培养的人成骨细胞，分析包括金属标本在内的临床材料的动态细胞行为，并建立评价此类材料生物亲和力的新方法。本研究揭示了以下数据。1)以人成骨细胞系(SAOS-2)为研究对象，从细胞铺展和细胞运动的角度分析了其在钛表面的行为，特别是表面光滑度(Li C，高S，小林N等)。Al.，2006)。此外，同一系统还检测了调节细胞运动的细胞内信号，表明肌动蛋白细胞骨架在Saos-2细胞的运动中起主要作用，细胞运动的方向被认为是由IP3-激酶信号调节的。2)利用人成骨细胞原代培养体系，建立了一种评价细胞在临床材料上运动性的新方法。当细胞被接种时，将一块玻璃盖子放在塑料培养皿的中央，盖子被重物压住。细胞贴在培养皿上后，取下盖子以获得无细胞的长方形区域，细胞开始迁移到该区域，允许测量细胞沿时间进程的运动性。在这种简单的方法之后，原代培养细胞的细胞运动性被证明具有显著的重复性。