Exosomal microRNA‐223 from neutrophil‐like cells inhibits osteogenic differentiation of PDLSCs through the cGMP‐PKG signaling pathway

Abstract

AbstractBackground and ObjectiveNeutrophils‐derived exosomes have been shown to cause tissue inflammation in many diseases, but their role in periodontitis, a neutrophil‐mediated disease, is unknown. Here, we investigated the effect of neutrophil‐like cells derived exosomes on osteogenic dysfunction of periodontal ligament stem cells (PDLSCs) in periodontitis.MethodsNeutrophil‐like cells were derived from HL‐60 cells by dimethylsulfoxide stimulation. Exosomes were isolated by ultracentrifugation and characterized using transmission electron microscopy, nanoflow cytometry and western blot. MicroRNA‐223 (miR‐223) expression were analyzed by real‐time PCR. Western blot, alkaline phosphatase (ALP), and alizarin red staining were conducted to assess whether exosomes could affect the osteogenic differentiation of PDLSCs. The expression of miR‐223 was inhibited in PDLSCs by transfecting with miR‐223 inhibitor. Cyclic guanosine monophosphate (cGMP) expression was determined by enzyme‐linked immunosorbent assay.ResultsWe found that miR‐223 was significantly increased in neutrophils and neutrophil‐like cells derived exosomes. Treatment with exosomes derived from neutrophil‐like cells upregulated miR‐223 expression and inhibited the osteogenic differentiation of PDLSCs, while transfection with miR‐223 inhibitor significantly promoted PDLSCs osteogenic differentiation. In addition, co‐treatment with KT5823, a cGMP–PKG pathway inhibitor, markedly abrogated the rescue effects of miR‐223 inhibitor on the osteogenic differentiation of PDLSCs.ConclusionsOur findings suggest that neutrophil‐like cells derived exosomes might inhibit osteogenic differentiation of PDLSCs by transporting miR‐223 and regulating the cGMP–PKG signaling pathway.