IADR Abstract Archives
Supracrestal Gingival Connective Tissue-Derived Mesenchymal Stem Cells Regulated Macrophage Cytokine Secretion
Objectives: To evaluate the effects of supracrestal gingival connective tissue-derived mesenchymal stem cells (SG-MSCs) on macrophage cell surface marker expression and cytokine expression.
Methods: SG-MSCs were isolated using enzymatic digestion from three healthy patients. MSCs characteristics comprising cell surface markers, colony-forming ability, and adipogenic and osteogenic differentiation potential were determined. THP-1 cells, a human monocyte cell line, were used to evaluate the effect of SG-MSCs on macrophage behavior. SG-MSCs were co-cultured with phorbol 12-myristate 13-acetate (PMA)-differentiated THP-1 cells directly in 6-well plates and indirectly in transwell plates at ratios of 0:1, 1:1 and 1:0. After 72 h, M1 and M2 cytokine markers (tumor necrosis factor-alpha (TNF-α) and interleukin (IL-10), respectively) were determined from culture supernatants by enzyme-linked immunosorbent assays. Macrophage CD80 (M1 marker) or CD206 (M2 marker) expression were also evaluated by flow cytometry after co-culture.
Results: MSCs from supracrestal gingival connective tissue expressed MSC markers, CD29, CD44, CD73, CD90, CD105, CD146, and STRO-1. SG-MSCs formed colonies and displayed osteogenic and adipogenic differentiation potential. SG-MSCs exhibited immunomodulatory effects on macrophage cytokine expression in both direct and indirect co-culture. Macrophages co-cultured with SG-MSCs produced significantly higher IL-10 levels (p<0.05) and significantly lower TNF-α levels (p<0.05) compared with macrophages cultured alone. However, changes in CD80 (M1 marker) nor CD206 (M2 marker) expression were not detected.
Conclusions: This study has demonstrated that SG-MSCs reduce TNF-α production while inducing IL-10 secretion from macrophages in co-culture, suggesting the potential of SG-MSCs in controlling inflammation of periodontitis and enhancing periodontal regeneration.
Methods: SG-MSCs were isolated using enzymatic digestion from three healthy patients. MSCs characteristics comprising cell surface markers, colony-forming ability, and adipogenic and osteogenic differentiation potential were determined. THP-1 cells, a human monocyte cell line, were used to evaluate the effect of SG-MSCs on macrophage behavior. SG-MSCs were co-cultured with phorbol 12-myristate 13-acetate (PMA)-differentiated THP-1 cells directly in 6-well plates and indirectly in transwell plates at ratios of 0:1, 1:1 and 1:0. After 72 h, M1 and M2 cytokine markers (tumor necrosis factor-alpha (TNF-α) and interleukin (IL-10), respectively) were determined from culture supernatants by enzyme-linked immunosorbent assays. Macrophage CD80 (M1 marker) or CD206 (M2 marker) expression were also evaluated by flow cytometry after co-culture.
Results: MSCs from supracrestal gingival connective tissue expressed MSC markers, CD29, CD44, CD73, CD90, CD105, CD146, and STRO-1. SG-MSCs formed colonies and displayed osteogenic and adipogenic differentiation potential. SG-MSCs exhibited immunomodulatory effects on macrophage cytokine expression in both direct and indirect co-culture. Macrophages co-cultured with SG-MSCs produced significantly higher IL-10 levels (p<0.05) and significantly lower TNF-α levels (p<0.05) compared with macrophages cultured alone. However, changes in CD80 (M1 marker) nor CD206 (M2 marker) expression were not detected.
Conclusions: This study has demonstrated that SG-MSCs reduce TNF-α production while inducing IL-10 secretion from macrophages in co-culture, suggesting the potential of SG-MSCs in controlling inflammation of periodontitis and enhancing periodontal regeneration.