Masi L, Franchi A, Santucci M, Danielli D, Arganini L, Giannone V, Formigli L, Benvenuti S, Tanini A, Beghè F
Department of Clinical Physiopathology, University of Florence Viale Pieraccini 6 50139, Italy.
Calcif Tissue Int. 1992 Sep;51(3):202-12. doi: 10.1007/BF00334548.
A number of studies have demonstrated the pivotal role of collagen molecules in modulating cell growth and differentiation. In order to analyze the direct effects of collagen type I on the osteoblastic phenotype, we have devised an in vitro culture system for studying the interactions between bovine collagen type I and Saos-2 cells, a human osteoblastic cell line. Saos-2 cells were cultured both on top of collagen-coated culture dishes as well as inside a three-dimensional collagen network. Plating on dishes treated with collagen induced maximal adhesion of Saos-2 cells after 24-hour incubation. Cells cultured on collagen gel matrix expressed about 2.5-fold more alkaline phosphatase when compared with untreated plastic dishes. On collagen-coated dishes the responsiveness of Saos-2 cells to parathyroid hormone was decreased, whereas no modifications were observed in the effect of vasoactive intestinal peptide on these cells. Using a microfluorimetric measurement of DNA, an increase of proliferation was observed in Saos-2 cells cultured on collagen gel. Saos-2 cells were also able to colonize collagen sponges and in this three-dimensional network they were able to synthesize osteocalcin, as assessed both by immunocytochemistry and radioimmunoassay. In this study we have demonstrated that bovine collagen type I exhibits favorable effects on attachment and functional and growth activities of a human osteoblastic cell line, encouraging its use as a bone graft material.
多项研究已证明胶原蛋白分子在调节细胞生长和分化中起关键作用。为了分析I型胶原蛋白对成骨细胞表型的直接影响,我们设计了一种体外培养系统,用于研究牛I型胶原蛋白与人类成骨细胞系Saos-2细胞之间的相互作用。Saos-2细胞既培养在胶原蛋白包被的培养皿上,也培养在三维胶原蛋白网络内部。接种在经胶原蛋白处理的培养皿上,孵育24小时后,Saos-2细胞的黏附达到最大值。与未处理的塑料培养皿相比,培养在胶原蛋白凝胶基质上的细胞碱性磷酸酶表达量高出约2.5倍。在胶原蛋白包被的培养皿上,Saos-2细胞对甲状旁腺激素的反应性降低,而血管活性肠肽对这些细胞的作用未观察到改变。通过微荧光法测量DNA,发现培养在胶原蛋白凝胶上的Saos-2细胞增殖增加。Saos-2细胞也能够在胶原蛋白海绵上定植,并且在这个三维网络中,通过免疫细胞化学和放射免疫测定评估,它们能够合成骨钙素。在本研究中,我们证明牛I型胶原蛋白对人类成骨细胞系的黏附、功能和生长活性具有有利影响,这促使其可作为骨移植材料使用。