Exosomes from TNF-α-treated human gingiva-derived MSCs enhance M2 macrophage polarization and inhibit periodontal bone loss

Yuki Nakao,Takao Fukuda,Qunzhou Zhang,Terukazu Sanui,Takanori Shinjo,Xiaoxing Kou,Chider Chen,Dawei Liu,Yukari Watanabe,Chikako Hayashi,Hiroaki Yamato,Karen Yotsumoto,Urara Tanaka,Takaharu Taketomi,Takeshi Uchiumi,Anh D Le,Songtao Shi,Fusanori Nishimura

doi:10.1016/j.actbio.2020.12.046

Abstract

Mesenchymal stem cell (MSC)–derived exosome plays a central role in the cell-free therapeutics involving MSCs and the contents can be customized under disease-associated microenvironments. However, optimal MSC-preconditioning to enhance its therapeutic potential is largely unknown. Here, we show that preconditioning of gingival tissue-derived MSCs (GMSCs) with tumor necrosis factor-alpha (TNF-α) is ideal for the treatment of periodontitis. TNF-α stimulation not only increased the amount of exosome secreted from GMSCs, but also enhanced the exosomal expression of CD73, thereby inducing anti-inflammatory M2 macrophage polarization. The effect of GMSC-derived exosomes on inflammatory bone loss were examined by ligature-induced periodontitis model in mice. Local injection of GMSC-derived exosomes significantly reduced periodontal bone resorption and the number of tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts, and these effects were further enhanced by preconditioning of GMSCs with TNF-α. Thus, GMSC-derived exosomes also exhibited anti-osteoclastogenic activity. Receptor activator of NF-κB ligand (RANKL) expression was regulated by Wnt5a in periodontal ligament cells (PDLCs), and exosomal miR-1260b was found to target Wnt5a-mediated RANKL pathway and inhibit its osteoclastogenic activity. These results indicate that significant ability of the TNF-α-preconditioned GMSC-derived exosomes to regulate inflammation and osteoclastogenesis paves the way for establishment of a therapeutic approach for periodontitis.

Highlights

Mesenchymal stem cells (MSCs) are multipotent progenitor cells that possess self-renewal with multiple differentiation capacities [1]
gingival tissuederived MSCs (GMSCs) were independently attached to the plastic culture dishes and to form colony forming unit-fibroblasts (CFU-Fs), which demonstrated that GMSCs exhibited greater clonal expansion and differentiation (Fig. 1A)
Since periodontal ligament (PDL)–alveolar bone interface plays a critical role in periodontal bone homeostasis [40], we examined the effect of GMSC-derived exosomes on RANKL/OPG signaling in PDL cells

Summary

Introduction

Mesenchymal stem cells (MSCs) are multipotent progenitor cells that possess self-renewal with multiple differentiation capacities [1]. Immunoregulatory effects of MSCs have contributed to clinical investigations for many diseases involving tissue inflammation and autoimmune disorders [2]. Such striking observations of MSCs exerting functional improvement without engraftment or differentiation [3] have led to the development of a therapeutic approach that utilizes the secretome of MSCs [4]. MSCs secretes multiple trophic factors including cytokines, growth factors and exosomes, all of which serve as paracrine mediators for immunoregulation and tissue regeneration [5]. The establishment of optical molecular-based protocol for MSCpreconditioning needs to be investigated

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Discussion

Conclusion