Stomatology department, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, No. 32 West Second Section, First Ring Road, Chengdu, 610072, Sichuan, China.
School of Medicine, University of Electronic Science and Technology of China, No. 4, Section 2, Jianshe North Road, Chengdu, 610054, Sichuan, China.
J Biol Inorg Chem. 2019 Aug;24(5):733-745. doi: 10.1007/s00775-019-01687-w. Epub 2019 Jul 19.
Researches have revealed the vital roles of the generated immune environment via the response of immune cells growing on biomaterial surfaces in the bone healing process. HAS and novel constructed microgrooved patterns of HAS (HAS-G) are widely used as biocompatible ceramic, especially as a mimic of the natural bone matrix. However, it is unclear whether osteoimmune response induced by HAS and HAS-G affects the osteogenic differentiation of bone marrow stromal cells (BMSCs). RAW264.7 cells were seeded on different surface of materials and cytokines released by macrophages were detected by enzyme-linked immunosorbent assay. The cell viability and mitochondrial function of macrophages seeded on different surface of materials were detected. Then, the effects of modified inflammatory microenvironment by macrophages on osteogenesis of BMSCs were measured by performing ALP staining, Alizarin Red S staining, and western blot. We confirmed that HAS-G is more favorable for RAW cell attaching and subsequently regulated the expression and release of cytokines/chemokines. Decrease in interleukin-6 (IL-6) release was further confirmed for contributing significantly to improve mitochondrial function in RAW cells. HAS-G-conditioned medium promoted osteogenic differentiation in BMSCs and was reversed by IL-6 addition. Decrease in IL-6 contributes to downregulation of miR-214 and subsequently upregulated p38/JNK pathway, which is potentially contributes to osteogenic promotion by HAS-G. This study is the first report to reveal the effects of HAS-G on osteogenesis via immune response, which could lead to a new insight into novel material for the advantage of biomaterials for tissue engineering applications.
研究揭示了免疫细胞在骨愈合过程中在生物材料表面生长时产生的免疫环境的重要作用。透明质酸（HAS）及其新型构建的微凹槽图案透明质酸（HAS-G）广泛用作生物相容性陶瓷，特别是作为天然骨基质的模拟物。然而，尚不清楚 HAS 和 HAS-G 诱导的骨免疫反应是否会影响骨髓基质细胞（BMSCs）的成骨分化。将 RAW264.7 细胞接种在不同材料的表面上，并通过酶联免疫吸附试验检测巨噬细胞释放的细胞因子。检测接种在不同材料表面的巨噬细胞的细胞活力和线粒体功能。然后，通过进行碱性磷酸酶染色、茜素红 S 染色和 Western blot 来测量巨噬细胞对 BMSCs 成骨作用的改性炎症微环境的影响。我们证实 HAS-G 更有利于 RAW 细胞附着，并随后调节细胞因子/趋化因子的表达和释放。白细胞介素 6（IL-6）释放的减少进一步证实有助于改善 RAW 细胞中的线粒体功能。HAS-G 条件培养基促进 BMSCs 的成骨分化，并且通过添加 IL-6 被逆转。IL-6 的减少有助于下调 miR-214，随后上调 p38/JNK 途径，这可能有助于 HAS-G 的成骨促进作用。这项研究首次揭示了 HAS-G 通过免疫反应对成骨的影响，这可能为组织工程应用中的新型材料提供新的见解。
