Comprehensive analysis of M2 macrophage-derived exosomes facilitating osteogenic differentiation of human periodontal ligament stem cells

Abstract

Background The role of periodontal ligament stem cells (PDLSCs) and macrophage polarization in periodontal tissue regeneration and bone remodeling during orthodontic tooth movement (OTM) has been well documented. Nevertheless, the interactions between macrophages and PDLSCs in OTM remain to be investigated. Consequently, the present study was proposed to explore the effect of different polarization states of macrophages on the osteogenic differentiation of PDLSCs. Methods After M0, M1 and M2 macrophage-derived exosomes (M0-exo, M1-exo and M2-exo) treatment of primary cultured human PDLSCs, respectively, mineralized nodules were observed by Alizarin red S staining, and the expression of ALP and OCN mRNA and protein were detected by RT-qPCR and Western blotting, correspondingly. Identification of differentially expressed microRNAs (DE-miRNA) in M0-exo and M2-exo by miRNA microarray, and GO and KEGG enrichment analysis of DE-miRNA targets, and construction of protein–protein interaction networks. Results M2-exo augmented mineralized nodule formation and upregulated ALP and OCN expression in PDLSCs, while M0-exo had no significant effect. Compared to M0-exo, a total of 52 DE-miRNAs were identified in M2-exo. The expression of hsa-miR-6507-3p, hsa-miR-4731-3p, hsa-miR-4728-3p, hsa-miR-3614-5p and hsa-miR-6785-3p was significantly down-regulated, and the expression of hsa-miR-6085, hsa-miR-4800-5p, hsa-miR-4778-5p, hsa-miR-6780b-5p and hsa-miR-1227-5p was significantly up-regulated in M2-exo compared to M0-exo. GO and KEGG enrichment analysis revealed that the downstream targets of DE-miRNAs were mainly involved in the differentiation and migration of multiple cells. Conclusions In summary, we have indicated for the first time that M2-exo can promote osteogenic differentiation of human PDLSCs, and have revealed the functions and pathways involved in the DE-miRNAs of M0-exo and M2-exo and their downstream targets.

M2 macrophages-derived exosomes regulate osteoclast differentiation by the CSF2/TNF-α axis

“…Moreover, M2-exos play an important role in bone formation and resorption balance. Previous studies have shown that M2-exos increase pro-inflammatory cytokines and suppress osteoblast differentiation of bone marrow mesenchymal stem cells and PDLSCs [ 27 , 28 ]. In addition, Chen et al [ 29 ] have reported that M2-exos impede bone marrow-derived macrophage osteoclast formation via the IL-10/IL-10R pathway, thereby reducing alveolar bone resorption.…”

Genetically engineered M2-like macrophage-derived exosomes for P. gingivalis-suppressed cementum regeneration: From mechanism to therapy

“…Due to their high permeability, low clearance, non-toxicity, therapeutic functions and the application of drug carriers, exosomes were widely studied for tissue regeneration [ 14 ]. Similar to M2 macrophages, M2-EXO was well-documented in inflammation targeting and elimination, angiogenesis, tumor growth, and bone formation [ 17 , [27] , [28] , [29] ]. However, its role in periodontal regeneration was rarely reported.…”

The Applications and Potentials of Extracellular Vesicles from Different Cell Sources in Periodontal Regeneration

“…However, most studies confirmed that M2-EVs could promote the osteogenesis of BMMSCs by delivering multiple miRNAs, such as miR-690 , miR-5106 and miR-26a-5p [ 80 , 82 , 83 ]. In terms of periodontal bone regeneration, Liao et al pointed out that M2-EVs might also promote the osteogenic differentiation of PDLSCs through a variety of miRNAs [ 84 ]. In addition to miRNAs, Chen et al found that M2-EVs were rich in IL-10, which promoted BMMSC osteogenic differentiation through IL-10/IL-10R signaling pathway, inhibited osteoclast formation, and reduced bone resorption in mice with periodontitis [ 56 ].…”