Department of Veterinary Clinical Sciences, Small Animal Clinic c/o Institute of Veterinary-Anatomy, -Histology and -Embryology, Justus-Liebig-University Giessen, Giessen, Germany.
Biomed Mater. 2017 Jul 4;12(4):045003. doi: 10.1088/1748-605X/aa6e29.
Herein, we aim to elucidate osteogenic effects of two silica-based xerogels with different degrees of bioactivity on human bone-derived mesenchymal stromal cells by means of scanning electron microscopy, quantitative PCR enhanced osteogenic effects and the formation of an extracellular matrix which could be ascribed to the sample with lower bioactivity. Given the high levels of bioactivity, the cells revealed remarkable sensitivity to extremely low calcium levels of the media. Therefore, additional experiments were performed to elucidate cell behavior under calcium deficient conditions. The results refer to capacity of the bone-derived stromal cells to overcome calcium deficiency even though proliferation, migration and osteogenic differentiation capabilities were diminished. One reason for the differences of the cellular response (on tissue culture plates versus xerogels) to calcium deficiency seems to be the positive effect of silica. The silica could be detected intracellularly as shown by time of flight-secondary ion mass spectrometry after cultivation of primary cells for 21 days on the surfaces of the xerogels. Thus, the present findings refer to different osteogenic differentiation potentials of the xerogels according to the different degrees of bioactivity, and to the role of silica as a stimulator of osteogenesis. Finally, the observed pattern of connexin-based hemichannel gating supports the assumption that connexin 43 is a key factor for calcium-mediated osteogenesis in bone-derived mesenchymal stromal cells.
在此,我们旨在通过扫描电子显微镜、定量 PCR 增强成骨效果和细胞外基质的形成来阐明两种具有不同生物活性程度的硅基干凝胶对人源骨髓间充质基质细胞的成骨作用,这可以归因于生物活性较低的样本。鉴于高生物活性,细胞对培养基中极低的钙水平表现出显著的敏感性。因此,进行了额外的实验来阐明细胞在缺钙条件下的行为。结果表明,骨髓基质细胞有能力克服钙缺乏,尽管增殖、迁移和成骨分化能力减弱。细胞对钙缺乏的反应(在组织培养板上与干凝胶上)存在差异的一个原因似乎是硅的积极作用。在用干凝胶表面培养原代细胞 21 天后,通过飞行时间二次离子质谱法可以在细胞内检测到硅。因此,本研究结果表明,根据不同的生物活性程度,干凝胶具有不同的成骨分化潜力,以及硅作为成骨刺激剂的作用。最后,观察到的连接蛋白基于半通道门控的模式支持这样的假设,即连接蛋白 43 是钙介导的骨髓间充质基质细胞成骨的关键因素。