WITHDRAWN: Human gingiva-derived mesenchymal stem cells promote osteogenic differentiation through their immunosuppressive function

Abstract

Human gingiva-derived mesenchymal stem cells (GMSCs) have emerged as a new MSC population exhibiting robust immune regulatory functions, multipotent differentiation potential, and regenerative ability. However, the effects of GMSCs on T cells remain unexplored. Herein, we aimed to evaluate whether GMSCs promote osteogenic differentiation by regulating immune cells. The GMSC phenotype was confirmed using the colony-forming assay, immunophenotyping, Oil red O staining, and Alizarin red staining. mRNA expression levels of proinflammatory molecules (interleukin-1β [IL-1β] and tumor necrosis factor-α [TNF-α]) and anti-inflammatory factors (IL-10) were measured by quantitative reverse-transcription PCR (qRT-PCR). Then, MC3T3-E1 cells were treated with the collected co-culture supernatant, followed by alkaline phosphatase (ALP) and immunofluorescence staining to evaluate osteogenic differentiation of MC3T3-E1 cells. qRT-PCR and western blotting were employed to analyze the expression levels of osteogenic differentiation proteins, including collagen type I (COL-1), ALP, osteopontin (OPN), and runt-related transcription factor 2 (RUNX2). GMSCs were successfully isolated and identified. We observed that GMSCs suppressed the activated T-cell function by downregulating IL-1β and TNF-α and upregulating IL-10. Simultaneously, the expression levels of osteogenesis-related genes (COL-1, ALP, OPN, and RUNX2) were markedly lower in the co-culture supernatant and Jurkat T cell supernatant groups than those in the normal culture medium group; however, expression levels were significantly increased in the co-culture supernatant group when compared with the Jurkat T cell supernatant group. Our findings indicate that GMSCs could promote the osteogenic differentiation of MC3T3-E1 cells by inhibiting the biological activity of activated T cells.