Institute for Biomedical Sciences, Interdisciplinary Cluster for Cutting Edge Research, Shinshu University, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan.
Biomedical Engineering Division, Graduate School of Medicine, Science and Technology, Shinshu University, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan.
Int J Mol Sci. 2021 Jul 20;22(14):7752. doi: 10.3390/ijms22147752.
Many biomaterials have been evaluated using cultured cells. In particular, osteoblast-like cells are often used to evaluate the osteocompatibility, hard-tissue-regeneration, osteoconductive, and osteoinductive characteristics of biomaterials. However, the evaluation of biomaterial osteogenesis-inducing capacity using osteoblast-like cells is not standardized; instead, it is performed under laboratory-specific culture conditions with different culture media. However, the effect of different media conditions on bone formation has not been investigated. Here, we aimed to evaluate the osteogenesis of MC3T3-E1 cells, one of the most commonly used osteoblast-like cell lines for osteogenesis evaluation, and assayed cell proliferation, alkaline phosphatase activity, expression of osteoblast markers, and calcification under varying culture media conditions. Furthermore, the various media conditions were tested in uncoated plates and plates coated with collagen type I and poly-L-lysine, highly biocompatible molecules commonly used as pseudobiomaterials. We found that the type of base medium, the presence or absence of vitamin C, and the freshness of the medium may affect biomaterial regeneration. We posit that an in vitro model that recapitulates in vivo bone formation should be established before evaluating biomaterials.
许多生物材料已经过培养细胞进行评估。特别是,成骨细胞样细胞通常用于评估生物材料的骨相容性、硬组织再生、骨传导性和骨诱导性特征。然而,使用成骨细胞样细胞评估生物材料成骨诱导能力尚未标准化;相反,它是在具有不同培养基的实验室特定培养条件下进行的。然而,不同的培养基条件对骨形成的影响尚未得到研究。在这里,我们旨在评估 MC3T3-E1 细胞的成骨作用,MC3T3-E1 细胞是最常用于成骨作用评估的成骨细胞系之一,并在不同的培养基条件下检测细胞增殖、碱性磷酸酶活性、成骨标志物的表达和钙化。此外,还在未涂层板和涂有胶原蛋白 I 和聚-L-赖氨酸的板上测试了各种培养基条件,胶原蛋白 I 和聚-L-赖氨酸是常用的高生物相容性假生物材料。我们发现基础培养基的类型、维生素 C 的存在与否以及培养基的新鲜度可能会影响生物材料的再生。我们假设在评估生物材料之前应该建立一个能够重现体内骨形成的体外模型。