After periodontal treatment, the local inflammatory environment surrounding periodontal tissues cannot be entirely eliminated. The means by which alveolar bone repair and regeneration are promoted in inflammatory environments have important clinical significance. As a powerful protein that promotes the differentiation of osteocytes, semaphorin 3A (Sema3A) shows potential for bone regeneration therapy. However, the effect of Sema3A on osteogenic differentiation in an inflammatory environment, as well as the underlying mechanism, have not yet been explored. We used lentivirus to transduce rat bone marrow-derived mesenchymal stem cells (rBMSCs) to stably overexpress Sema3A. Lipopolysaccharide from Escherichia coli (E. coli LPS) was used to stimulate rBMSCs to establish an inflammatory environment. ALP staining, Alizarin red staining, ALP activity tests, quantitative RT-PCR (qRT-PCR), and Western blotting were used to elucidate the effect of Sema3A on the osteogenesis of rBMSCs in inflammatory environments. XAV939 and LiCl were used to determine whether the Wnt/β-catenin signaling pathway was involved in attenuating the inhibition of Sema3A-induced osteogenic differentiation by LPS. The qRT-PCR and Western blot results demonstrated that the lentiviral vector (LV-NC) and lentiviral-Sema3A (LV-Sema3A) were successfully transduced into rBMSCs. An inflammatory environment could be established by stimulating rBMSCs with 1 μg/ml E. coli LPS. After Sema3A overexpression, mineral deposition was exacerbated, and the BSP and Runx2 gene and protein expression levels were increased. Furthermore, E. coli LPS activated the Wnt/β-catenin signaling pathway and decreased rBMSC osteogenesis, but these effects were attenuated by Sema3A. In conclusion, Sema3A could protect BMSCs from LPS-mediated inhibition of osteogenic differentiation in inflammatory environments by suppressing the Wnt/β-catenin pathway.

牙周治疗后，牙周组织周围的局部炎症环境并不能完全消除。在炎症环境中促进牙槽骨修复和再生的手段具有重要的临床意义。作为一种促进骨细胞分化的强大蛋白质，信号素 3A (Sema3A) 显示出用于骨再生治疗的潜力。然而，尚未探索 Sema3A 在炎症环境中对成骨分化的影响，以及潜在的机制。我们使用慢病毒转导大鼠骨髓间充质干细胞 (rBMSCs) 以稳定过表达 Sema3A。来自大肠杆菌（E.coli ）的脂多糖LPS) 用于刺激 rBMSCs 以建立炎症环境。ALP 染色、茜素红染色、ALP 活性测试、定量 RT-PCR (qRT-PCR) 和蛋白质印迹用于阐明 Sema3A 在炎症环境中对 rBMSCs 成骨的影响。XAV939 和 LiCl 用于确定 Wnt/β-catenin 信号通路是否参与减弱 LPS 对 Sema3A 诱导的成骨分化的抑制作用。qRT-PCR 和蛋白质印迹结果表明，慢病毒载体 (LV-NC) 和慢病毒-Sema3A (LV-Sema3A) 已成功转导至 rBMSCs。可以通过用 1 μg/ml大肠杆菌刺激 rBMSCs 来建立炎症环境脂多糖。Sema3A过表达后，矿物质沉积加剧，BSP和Runx2基因及蛋白表达水平升高。此外，大肠杆菌LPS 激活 Wnt/β-catenin 信号通路并减少 rBMSC 成骨，但这些作用被 Sema3A 减弱。总之，Sema3A 可以通过抑制 Wnt/β-catenin 通路来保护 BMSCs 免受 LPS 介导的炎症环境中成骨分化的抑制。