Objectives: Human periodontal ligament stem cells (hPDLSCs) bear multilineage differentiation potential and represent the cytological basis of periodontal tissue regeneration. microRNA (miR) is accepted as a critical regulator of cell differentiation. This study explored the molecular mechanism of miR-200a-3p in osteogenesis of hPDLSCs.
Material and methods: hPDLSCs were cultured and identified in vitro. miR-200a-3p expression during osteogenic differentiation of hPDLSCs was detected. hPDLSCs were transfected with miR-200a-3p mimic or miR-200a-3p inhibitor. Alkaline phosphatase (ALP) activity, calcified nodules and osteogenesis-related genes of hPDLSCs were measured. The binding relationship between miR-200a-3p and ZEB2 was predicted and verified. hPDLSCs were infected with sh-ZEB2, and then the osteogenic capacity was examined. miR-200a-3p inhibitor-transfected hPDLSCs were infected with sh-ZEB2. The key proteins of the NF-κB pathway were measured.
Results: miR-200a-3p expression was downregulated during osteogenic differentiation of hPDLSCs. Upregulation of miR-200a-3p reduced ALP activity, calcified nodules and osteogenesis-related genes of hPDLSCs, while downregulation of miR-200a-3p facilitated the osteogenesis of hPDLSCs. miR-200a-3p targeted ZEB2. ZEB2 silencing repressed osteogenesis of hPDLSCs. ZEB2 silencing attenuated the promoting effect of miR-200a-3p inhibitor on osteogenesis of hPDLSCs. miR-200a-3p activated the NF-κB pathway by targeting ZEB2.
Conclusion: miR-200a-3p repressed osteogenesis of hPDLSCs by targeting ZEB2 and activating the NF-κB pathway. This study may offer insights for periodontal tissue regeneration engineering.
Keywords: NF-κB pathway; Osteogenesis; ZEB2; human periodontal ligament stem cells; miR-200a-3p; molecular mechanism.