Elsevier

Pharmacological Research

Volume 147, September 2019, 104381
Pharmacological Research

MOTS-c inhibits Osteolysis in the Mouse Calvaria by affecting osteocyte-osteoclast crosstalk and inhibiting inflammation

https://doi.org/10.1016/j.phrs.2019.104381Get rights and content

Abstract

The Mitochondrial-derived peptide MOTS-c has recently been reported as a 16-amino acid peptide regulating metabolism and homeostasis in different cells. However, its effects on immune cells and bone metabolism are rarely reported. Here we demonstrate that MOTS-c treatment in ultra-high molecular weight polyethylene (UHMWPE) particle-induced osteolysis mouse model alleviated bone erosion and inflammation. MOTS-c increased osteoprotegerin (OPG)/ receptor activator of nuclear factor kappa-B ligand (RANKL) ratio in osteocytes, leading to inhibition of osteoclastogenesis. In primary bone marrow macrophages (BMMs) MOTS-c alleviated STAT1 and NF-κB phosphorylation triggered by UHMWPE particles. Promoting ROS production or suppressing peroxisome proliferator-activated receptor γ (PPARγ) coactivator-1α (PGC-1α) by adenosine 5‘-monophosphate (AMP)-activated protein kinase (AMPK) repression blocked these anti-inflammatory effects of MOTS-c treatment. Taken together, these findings provide evidence that the small peptide inhibits osteoclastogenesis by regulating osteocyte OPG/RANKL secretion and suppressing inflammation via restraining NF-κB and STAT1 pathway. Moreover, its effects on NF-κB activation is dependent on the AMPK-PGC-1α-ROS axis, suggesting its potential use in osteolysis and other inflammation disorders.

Introduction

UHMWPE wear-induced osteolysis is a complex process during which prosthetic debris released from the surface of prosthetic joints made of wear-producing-material induces an immune response that favors bone catabolism, thereby resulting in the loosening of the prosthesis, fracture, and other failures [1]. Although many factors account for the process, bone resorption mediated by osteoclasts and the self-propagating immune response initiated by mononuclear macrophages are the main effectors. Schmalzried et al. examined histological samples derived from areas of bone resorption from patients with radiographic evidence of periprosthetic osteolysis and detected polyethylene debris-laden macrophages [2]. Further evidence confirmed the involvement of UHMWPE wear particles in aseptic inflammation [3]. For instance, particulates with diameters of 0.1–1.0 μm can be phagocytosed by macrophages and are the most biologically active, triggering TNF-α, IL-6, IL-1β, and prostaglandin E2 (PGE2) production and causing local infiltration, which promotes osteoclastogenesis and bone resorption and disturbs bone metabolism by inducing RANKL secretion and suppressing that of its decoy, OPG [[4], [5], [6]]. Therefore, studies on therapeutic agents for osteolysis focus on those utilized in catabolic bone diseases such as bisphosphonates [7], TNF-α antagonists, IL-1β antagonists [8], IL-10, OPG vectors [9], and the recently approved monoclonal antibody against RANKL, denosumab [10]. However, none of these compounds can simultaneously block both the main effects of wear debris and the side effects form barriers to a wider use [11]. Here, we identify an mtDNA-encoded peptide, MOTS-c, that can alleviate inflammation and bone resorption in a commonly utilized osteolysis model induced by UHMWPE particles [12] and report that it may be a potential drug for aseptic loosening.

MOTS-c, a mitochondrial-encoded peptide, has been characterized as a 16-amino acid peptide with multiple roles in cell metabolism and Lee et al. first identified the peptide, which was encoded in short open reading frames within the mitochondrial genome, and showed that it promoted metabolic homeostasis and reduced insulin resistance [13]. Kim et al. discovered that the peptide dynamically translocated to the nucleus in response to metabolic stress, where it regulated the expression of nuclear genes [14]. Recently, several studies on the peptide have revealed its role in adipose homeostasis, infectious disease, and endothelial dysfunction [[15], [16], [17]]. However, it is not known whether MOTS-c can affect immune cells during aseptic inflammation. In search of novel drugs for aseptic loosening with fewer side effects and better therapeutic outcomes, we set out to explore the anti-inflammatory effects and bone-protective properties of MOTS-c and to define the mechanism of its anti-inflammatory effects.

Section snippets

Cell culture and animal experiment

Dulbecco’s modified Eagle medium (Gibco, USA) supplemented with 10% fetal bovine serum (Gibco, USA), penicillin (100 U/mL, Gibco), and streptomycin (100 mg/ml) (Gibco) was used to culture primary mouse bone marrow macrophages (BMMs) and primary mouse osteocytes. Cells were obtained as previously described [18]. Osteocytes were obtained from mouse calvaria cultured in osteogenic medium supplemented with 1 nM dexamethasone, 50 μM ascorbic acid-2-phosphate and 20 mM β-glycerophosphate

MOTS-c protects bone mass and alleviates inflammation in UHMWPE particle-induced osteolysis

To validate the effects of MOTS-c on UHMWPE particle-induced osteolysis, we conducted experiments using an osteolysis mouse model as previously described [19]. Briefly, UHMWPE particles were used to simulate periprosthetic aseptic inflammation in vivo. We then evaluated bone mass in different groups with micro-CT scanning. The micro-CT scanning results confirmed UHMWPE particle-induced bone erosion, and multiple bone defects were detected in the control group treated with the UHMWPE particles.

Discussion

According to one study on primary total hip arthroplasties (THA), one of the most common causes for total hip revision surgery is aseptic loosening [41]. Aseptic loosening by periprosthetic osteolysis is the leading cause of failure after total joint arthroplasty (TJA), with the reported incidence rate ranging from 10% to 70% and the incidence of TJA still increasing [[42], [43]]. Wear particles cause inflammation and osteolysis, thereby leading to continuous synovitis which causes pain,

Conclusion

In conclusion, considering its constitutive role in mitochondrial protein expression, our current data suggest that MOTS-c has a potent anti-inflammatory effect in osteolysis therapy with fewer side effects, and it holds promise in anti-inflammatory therapy in the future.

Ethics statement

This study was carried out in accordance with the recommendations of the Laboratory Animal Center of Fourth Military Medical University (permit no. 20161003).

CRediT authorship contribution statement

Zhao Yan: Experiment design, Writing - review & editing. Shu Zhu: Data analysis, Investigation. Hanli Wang: Methodology, Investigation. Li Wang: Conceptualization, Investigation. Tianshu Du: Investigation. Zichen Ye: Investigation. Dongsheng Zhai: Investigation. Zheng Zhu: Data analysis. Xiaoxi Tian: Funding acquisition. Zifan Lu: Funding acquisition, Experiment design. Xiaorui Cao: Project administration, Funding acquisition.

Declaration of Competing Interest

The authors declare that they have no conflict of interest.

Acknowledgements

This work was supported by the National Natural Science Foundation of China (81501936), (31571215) and (81802168).

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    These authors have contributed equally to this work.

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