Sun Xiaoyu, Su Wen, Ma Xiaomin, Zhang Huaiying, Sun Zhe, Li Xudong
National Engineering Research Center for Biomaterials, Sichuan University, Chengdu 610064, P.R. China.
Regen Biomater. 2018 Mar;5(2):93-103. doi: 10.1093/rb/rbx018. Epub 2017 Jun 30.
Collagen (COL), collagen/hydroxyapatite (COL/HA), HA and biphasic calcium phosphate were prepared as representative bone grafting materials with composition analogous to bone, and their structural characteristics were analyzed. The rat bone mesenchymal stem cells (BMSCs) were further seeded onto four groups of materials, and BMSCs grown in basic medium and standard osteogenic medium were set as controls of a reference model to show the basic and osteogenic behavior of cells without the intervention of materials. Cellular behaviors were characterized, including proliferation, spreading morphology and expression of osteogenesis factors. The rat BMSCs proliferated properly with time on four groups of materials as well on two groups of controls, and typical cuboidal, polygonal and extremely-elongated morphologies of cells were observed. According to the real-time polymerase chain reaction data, a higher osteogenic gene expression level was dependent upon the growing morphology but not the proliferation rate of cells, and the osteogenic differentiation capacity of cells onto four groups of materials varied in specific genes. In general, BMSCs exhibited the highest osteogenic capacity onto COL/HA, but the poorest onto HA. The growing behaviors of cells on materials were further discussed in comparison with the cases of OC and BC of the reference model. The present attempt to comparatively analyze cell experimental data with a reference model is expected to be useful for revealing the difference in the osteogenic capability of MSCs onto materials or even the bioactivity of materials.
制备了胶原蛋白(COL)、胶原/羟基磷灰石(COL/HA)、羟基磷灰石(HA)和双相磷酸钙作为具有与骨类似组成的代表性骨移植材料,并分析了它们的结构特征。将大鼠骨髓间充质干细胞(BMSCs)进一步接种到四组材料上,并将在基础培养基和标准成骨培养基中生长的BMSCs作为参考模型的对照,以显示无材料干预时细胞的基本和成骨行为。对细胞行为进行了表征,包括增殖、铺展形态和成骨因子的表达。大鼠BMSCs在四组材料以及两组对照上均随时间正常增殖,并观察到典型的立方形、多边形和极度伸长的细胞形态。根据实时聚合酶链反应数据,较高的成骨基因表达水平取决于细胞的生长形态而非增殖速率,并且细胞在四组材料上的成骨分化能力在特定基因中有所不同。总体而言,BMSCs在COL/HA上表现出最高的成骨能力,但在HA上表现最差。与参考模型的OC和BC情况相比,进一步讨论了细胞在材料上的生长行为。目前用参考模型比较分析细胞实验数据的尝试有望有助于揭示间充质干细胞在材料上的成骨能力差异甚至材料的生物活性差异。