Diomede F, Merciaro I, Martinotti S, Cavalcanti M F X B, Caputi S, Mazzon E, Trubiani O
Laboratory of Stem Cells and Regenerative Medicine, University “G. d’Annunzio”, Chieti-Pescara, Chieti, Italy.
Department of Pharmacy, University “G. d’Annunzio”; Chieti-Pescara, Chieti, Italy.
J Biol Regul Homeost Agents. 2016 Oct-Dec;30(4):1009-1018.
miR-2861 endorsing osteoblast differentiation through the overexpression of Runt-related transcription factor 2 (RUNX2) protein has been recently described. In this study we evaluated: the performance of living construct, composed by human Periodontal Ligament Stem Cells (hPDLSCs) and 3D scaffold (EXg), and the behaviour of miR-2861/RUNX2 expression pathway on the osteogenic commitment. Human PDLSCs were seeded with and without EXg scaffold and cultured under basal and osteogenic conditions. Morphological features, adhesiveness and differentiation abilities were analysed using scanning electron and confocal laser scanning microscopy. Time-course of RUNX2, ALP, OPN and miR-2861 were evaluated through RT-PCR analysis. Our results highlighted that the osteogenic differentiation was mostly obvious in the hPDLSCs, grown onto 3D scaffold in presence of osteoinductive medium. Moreover, the overexpression of miR-2861 and RUNX2 in hPDLSCs cultured in presence of EXg under osteogenic and standard conditions was demonstrated. In synthesis, the increased expression of miR-2861/RUNX2 provides new insights regarding miRNA signaling network in the presence of scaffold providing an additional method to evaluate the performance of biomaterial in bone regeneration.
最近有研究描述了通过过表达 runt 相关转录因子 2(RUNX2)蛋白来支持成骨细胞分化的 miR - 2861。在本研究中,我们评估了:由人牙周膜干细胞(hPDLSCs)和 3D 支架(EXg)组成的活性构建体的性能,以及 miR - 2861/RUNX2 表达途径在成骨过程中的行为。将人 PDLSCs 接种在有和没有 EXg 支架的情况下,并在基础和成骨条件下培养。使用扫描电子显微镜和共聚焦激光扫描显微镜分析形态特征、粘附性和分化能力。通过 RT - PCR 分析评估 RUNX2、ALP、OPN 和 miR - 2861 的时间进程。我们的结果表明,在存在骨诱导培养基的情况下,在 3D 支架上生长的 hPDLSCs 中,成骨分化最为明显。此外,还证明了在成骨和标准条件下,在存在 EXg 的情况下培养的 hPDLSCs 中 miR - 2861 和 RUNX2 的过表达。总之,miR - 2861/RUNX2 表达的增加为在存在支架的情况下 miRNA 信号网络提供了新的见解,为评估生物材料在骨再生中的性能提供了一种额外的方法。