Semaphorin3A has been identified as a potent osteoprotective factor that simultaneously inhibits bone resorption and promotes bone formation. The present study demonstrates the effect of the overexpression of Sema3A on the proliferation and differentiation of rat gingival mesenchymal stem cells (GMSCs) in the lipopolysaccharide (LPS)-induced inflammatory environment. rGMSCs were transfected with viral stocks of pLenO-GTP-Sema3A (Lv-Sema3A group) or pLenO-GTP (Lv-NC group), with rGMSCs as a control. The transfection efficiency was determined by flow cytometry. Cell proliferation was assessed using a Cell Counting Kit-8 assay. The expressions of alkaline phosphatase (ALP), osteocalcin (OCN), and runt-related transcription factor 2 (Runx2) were determined at 3, 7 and 14 days after the osteogenic induction culture with or without LPS using real-time PCR and Western blot. Alizarin Red staining was performed at 28 days. A pLenO-GTP-Sema3A-mediated transfection of rGMSC stably overexpressing Sema3A was built up. The overexpression of Sema3A promoted cell proliferation in the LPS-induced inflammatory environment. In addition, osteogenesis-related genes were upregulated in the Lv-Sema3A group compared with the control group. Also, after LPS administration, the overexpression of Sema3A enhanced the expression of the osteogenic genes in the LPS-induced inflammatory environment. Hence, Sema3A gene-modified rGMSCs show better osteogenic differentiation and proliferation capacities compared with rGMSCs in the LPS-induced inflammatory environment.
Keywords: Semaphorin3A; bone tissue engineering; gingival mesenchymal stem cells; lipopolysaccharide; transfection.
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