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

本研究旨在通过体外培养人成骨细胞，分析其在临床材料（包括金属材料）上的动态细胞行为，建立评价材料生物亲和性的新方法。本研究揭示了以下数据。1)通过使用成骨细胞的人细胞系（Saos-2），从细胞铺展和细胞运动的角度分析它们在钛表面上的行为，特别参考表面光滑度（Li C，Gao S，小林N，et.例如，2006年）。此外，细胞内信号调节细胞运动性测定相同的系统，显示肌动蛋白细胞骨架在Saos-2细胞的细胞运动性的主要作用，细胞运动的方向被认为是由IP 3-激酶信号调节。2)利用人成骨细胞原代培养体系，建立了一种新的评价细胞在临床材料上运动能力的方法。当接种细胞时，将玻璃盖玻片置于塑料培养皿的中心，并用重物按压盖玻片。细胞附着在培养皿上后，除去盖玻片以获得无细胞的矩形区域，细胞开始迁移到该区域中，从而允许测量沿着时间进程的细胞运动性。在这种简单的方法之后，原代培养细胞的细胞运动性显示出以显著的再现性进行分析。

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.