Mitochondria-rich extracellular vesicles derived from the culture supernatant of human synovial Fluid-derived mesenchymal stem cells Inhibited senescence of Stressed/inflammatory Licensed chondrocytes and Delayed Osteoarthritis progression.

Abstract

Background: Mitochondrial dysfunction induces chondrocyte senescence, thereby precipitating articular cartilage (AC) degeneration in the pathogenesis of osteoarthritis (OA). Although the transfer of mitochondria from mesenchymal stem cells (MSCs) to host cells and their potential protective role have been demonstrated, whether MSCs can alleviate chondrocyte mitochondrial dysfunction or reverse OA progression remains unclear.

Methods: A mitochondrial tracer was used to investigate the transfer of mitochondria-rich extracellular vesicles (MEV) derived from the culture supernatant of human synovial fluid-derived mesenchymal stem cells (hSF-MSCs). Human articular chondrocytes (hACs) impaired by oxidative stress co-incubated with MEV were used for experimental research in vitro. Healthy hACs and stressed hACs were cultured separately acting as the control groups. The MEV was injected into the OA rats' knee joint serving as experimental group. Healthy and OA rats were served as the control groups. Quantitative reverse transcription polymerase chain reaction (qRT-PCR), western blot (WB), enzyme- linked immunosorbent assay (ELISA), flow cytometry (FC), immunofluorescence (IF), fluorescence spectrophotometer (FS), immunohistochemistry (IHC) and other methods are used to analyze the effect of MEV on hACs and OA progression.

Results: MEV derived from hSF-MSCs could transfer into hACs. Compared to the negative control group, co-incubation with MEV resulted in a significant down-regulation of oxidative stress markers and senescence-associated proteins in hACs, while improved mitochondrial function of hACs. Moreover, the MEV could traverse the dense interstitial layer and migrate towards the deeper cartilage, while intra-articular injection of MEV could effectively attenuate AC degeneration.

Conclusion: The transfer of MEV derived from hSF-MSCs represents a promising strategy for safeguarding AC, thereby offering a potential avenue and mechanism for the treatment of OA.