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Differential miRNA expression of healthy and inflammatory periodontal ligament stem cells-derived exosomes

ABSTRACT: Differential miRNA expression of healthy and inflammatory periodontal ligament stem cells-derived exosomes

PROVIDER: PRJNA686378 | ENA |

REPOSITORIES: ENA

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Differential miRNA expression of healthy and inflammatory periodontal ligament stem cells-derived exosomes

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Project description:Objective: To investigate the effects of stretch-induced periodontal ligament cell (PDLC) exosomes on osteoblast differentiation, and to explore their regulatory role in mechanical force-related periodontal tissue remodeling.Design: After applying 20% stretch loading to human periodontal ligament cells, exosomes were extracted from the supernatant and co-cultured with osteoblasts to detect their effects on osteogenic differentiation. Meanwhile, the exosomes were sequenced by high-throughput microRNA sequencing for bioinformatic analysis and validation to explore exosome signaling pathways through miRNAs.Results: Stretch-induced PDLC exosomes could be taken up by osteoblasts and promoted osteogenic differentiation of osteoblasts, as demonstrated by the increased expression levels of osteogenesis-related factors and enhanced ALP staining. In the miRNA profile of differentially expressed PDLC exosomes under stretch loading, miRNA-181d-5p was up-regulated significantly. The expression levels of osteogenesis-related factors and ALP staining were also increased in osteoblasts transfected with miR-181d-5p, and this effect might be related to the inhibitory role of exosomal miR-181d-5p on tumor necrosis factor (TNF).Conclusions: Stretch-induced PDLC exosomes exhibited a promoting effect on osteogenic differentiation, which might result from the inhibition of TNF via exosomal miR-181d-5p.

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Project description:Maresin-1 (MaR1) and Resolvin E1 (RvE1) are specialized pro-resolving lipid mediators (SPMs) that regulate inflammatory processes. We have previously demonstrated the hard and soft tissue regenerative capacity of RvE1 in an in vivo model of periodontal disease characterized by inflammatory tissue destruction. Regeneration of periodontal tissues requires a well-orchestrated processes mediated by periodontal ligament stem-cells. However, limited data are available on how SPMs can regulate the regenerative properties of human periodontal ligament stem cells (hPDLSCs) under inflammatory conditions. Thus, we measured the impact of MaR1 and RvE1 in an in vitro model of hPDLSCs after stimulation with IL-1β and TNF-α by evaluating pluripotency, migration, proliferation, cell death, periodontal ligament markers (α-smooth muscle actin, tenomodulin, and periostin), cementum-osteogenic differentiation and phosphoproteomic perturbations. The data showed that the inflammatory milieu suppresses pluripotency, proliferation and migration of hPDLSCs; MaR1 and RvE1 both restored regenerative capacity by increasing hPDLSC proliferation, accelerating wound healing/migration, and upregulating periodontal ligament markers and cementum-osteogenic differentiation. Protein phosphorylation perturbations were associated with the SPM-induced regenerative capacity of hPDLSCs. Together, these results demonstrate that MaR1 and RvE1 restore or improve the regenerative properties of highly specialized stem cells when inflammation is present and offer opportunities for direct pharmacologic treatment of lost tissue integrity.

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