Department of Medical, Oral and Biotechnological Sciences, University "G. d'Annunzio" Chieti-Pescara, 66100 Chieti, Italy.
ASL02 Lanciano-Vasto-Chieti, Chieti "Ss. Annunziata" Hospital, 66100 Chieti, Italy.
Int J Mol Sci. 2018 Dec 6;19(12):3916. doi: 10.3390/ijms19123916.
The aim of the present research was the evaluation of the behavior of human periodontal ligament stem cells (hPDLSCs), cultured in presence of Endobon Xenograft Granules (G), a fully deproteinated hydroxyapatite ceramic scaffold derived from cancellous bovine bone. hPDLSCs were seeded with and without G for 24 h to 1 week. The cell growth, morphological features, adhesiveness, differentiation ability, modulation of miR-210 and Vascular Endothelial Growth Factor (VEGF) secretion were analyzed by means of MTT assay, Scanning Electron Microscopy (SEM), Confocal Laser Scanning Microscopy (CLSM), Alizarin Red S assay, RT-PCR and ELISA test, respectively. hPDLSCs grown on the biomaterial showed the ability to form focal adhesion on the substrate, as demonstrated by vinculin expression. These data were supported by SEM analysis showing that an adhesiveness process associated to cell growth occurs between cells and biomaterials. The osteogenic differentiation, evaluated by morphological, biochemical, and RT-PCR analysis, was pronounced in the hPDLSCs grown in the three-dimensional inorganic bovine bone substitute in the presence of osteoinductive conditions. In addition, an upregulation of miR-210 and VEGF was evident in cells cultured in presence of the biomaterial. Our results inspire us to consider granules not only an adequate biocompatible three-dimensional biomaterial, but also an effective inductor of miR-210 and VEGF; in fact, the involvement of miR-210 in VEGF secretion could offer a novel regulatory system in the early steps of the bone-regeneration process.
本研究旨在评估人牙周膜干细胞（hPDLSCs）在 Endobon Xenograft 颗粒（G）存在下的行为，G 是一种源自松质牛骨的完全去蛋白羟磷灰石陶瓷支架。将 hPDLSCs 与 G 共培养 24 小时至 1 周。通过 MTT 测定、扫描电子显微镜（SEM）、共聚焦激光扫描显微镜（CLSM）、茜素红 S 测定、RT-PCR 和 ELISA 试验分别分析细胞生长、形态特征、黏附性、分化能力、miR-210 和血管内皮生长因子（VEGF）分泌的调节。在生物材料上生长的 hPDLSCs 表现出在底物上形成焦点黏附的能力，这由 vinculin 表达证明。SEM 分析支持了这一数据，表明细胞与生物材料之间发生了与细胞生长相关的黏附过程。在存在成骨诱导条件下，通过形态学、生化和 RT-PCR 分析评估的成骨分化在三维无机牛骨替代物中生长的 hPDLSCs 中明显。此外，在存在生物材料的情况下培养的细胞中，miR-210 和 VEGF 的表达明显上调。我们的结果使我们不仅将颗粒视为一种合适的生物相容性三维生物材料，而且还将其视为 miR-210 和 VEGF 的有效诱导物；事实上，miR-210 在 VEGF 分泌中的参与可能为骨再生过程的早期步骤提供了一种新的调节系统。
