Osteogenic differentiation of dental pulp stem cells (DPSCs) is considered as a promising strategy in posterior maxilla tooth implantation. Information on the function and mechanisms of long non-coding RNAs (lncRNAs) in osteogenic differentiation of DPSCs is growing, however, the mechanism of LINC00968 and miR-3658 in regulating osteogenic differentiation of DPSCs still needs to be explored. In this study, the LINC00968 and miR-3658 expression level was upregulated and downregulated in DPSCs and peri-implantitis DPSCs (pDPSCs) treated with bone morphogenic protein (BMP)2, respectively. Moreover, the effects of LINC00968 and miR-3658 on BMP2-induced osteogenic differentiation of DPSCs in vitro using Alizarin Red S staining, alkaline phosphatase (ALP) activity, quantitative real time PCR and Western blot assays showed that overexpression of LINC00968 significantly promoted mineralized bone matrix, alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2), and osterix (OSX) expression levels for osteogenic differentiation of DPSCs and pDPSCs; and overexpression of miR-3658 showed an opposite result that inhibited osteogenic differentiation of DPSCs and pDPSCs. Luciferase reporter assay showed that luciferase activities of LINC00968-WT reporter and RUNX2-WT reporter were strongly suppressed by miR-3658 overexpression. In addition, the miR-3658 upregulation interfered ectopic bone formation in vivo stimulated by LINC00968. In general, we had identified a novel molecular pathway involving LINC00968/miR-3658/RUNX2 during DPSCs and pDPSCs differentiation into osteoblasts, which might facilitate bone anabolism.

牙髓干细胞（DPSCs）的成骨分化被认为是上颌后牙种植的一种有前景的策略。关于长链非编码 RNA (lncRNAs) 在 DPSCs 成骨分化中的功能和机制的信息越来越多，但是，LINC00968 和 miR-3658 调节 DPSCs 成骨分化的机制仍有待探索。在本研究中，LINC00968 和 miR-3658 的表达水平分别在用骨形态发生蛋白 (BMP)2 处理的 DPSCs 和种植体周围炎 DPSCs (pDPSCs) 中上调和下调。此外，LINC00968和miR-3658对BMP2诱导的DPSCs体外成骨分化的影响使用茜素红 S 染色、碱性磷酸酶 (ALP) 活性、实时定量 PCR 和蛋白质印迹分析表明，LINC00968 的过表达显着促进矿化骨基质、碱性磷酸酶 (ALP)、矮小相关转录因子 2 (RUNX2) 和 osterix (OSX) DPSCs 和 pDPSCs 成骨分化的表达水平；miR-3658 的过表达显示出相反的结果，即抑制 DPSCs 和 pDPSCs 的成骨分化。荧光素酶报告基因检测表明，LINC00968-WT 报告基因和 RUNX2-WT 报告基因的荧光素酶活性被 miR-3658 过表达强烈抑制。此外，miR-3658的上调干扰了体内异位骨形成由 LINC00968 刺激。总的来说，我们在 DPSCs 和 pDPSCs 分化为成骨细胞期间确定了一个新的分子途径，涉及 LINC00968/miR-3658/RUNX2，这可能促进骨合成代谢。