Mitochondria-Rich Extracellular Vesicles From Bone Marrow Stem Cells Mitigate Muscle Degeneration in Rotator Cuff Tears in a Rat Model Through Macrophage M2 Phenotype Conversion.

PURPOSE: To investigate the protective effects of extracellular vesicles derived from bone marrow stem cells (BMSC-EVs) on muscle degeneration in a rat model of rotator cuff tendon and suprascapular nerve (SSN) transection (termed the RCT-SSN model), focusing on mitochondrial transfer.

METHODS

The EVs were identified and characterized. The RCT-SSN model was established by transecting the supraspinatus, infraspinatus tendons, and suprascapular nerve. Ninety-six rats were divided into 4 groups (n = 24 each): sham surgery, RCT-SSN treated with BMSC-EVs, RCT-SSN treated with EVs from rhodamine 6G-pretreated BMSCs (Rho-EVs), or phosphate-buffered saline. Intramuscular injections were administered every 2 weeks. After 12 weeks, supraspinatus muscles were analyzed for atrophy, fibrosis, oxidative stress, macrophage phenotypes, serum cytokines, and mitochondrial characteristics. In vitro experiments included EVs tracking in macrophages, macrophage phenotype characterization, and inflammatory cytokine profiling.

RESULTS

BMSC-EVs and Rho-EVs displayed similar morphology, but only BMSC-EVs contained functional mitochondria. BMSC-EVs significantly reduced muscle weight loss (0.047 ± 0.010% vs 0.145 ± 0.013%, P < .001), increased muscle fiber cross-sectional area (2037 ± 231.9 μm2 vs 527.9 ± 92.01 μm2, P < .001), and decreased fibrosis (12.09 ± 3.31% vs 25.69 ± 4.84%, P < .001) compared with phosphate-buffered saline. BMSC-EVs enhanced superoxide dismutase activity (93.3 ± 11.8 U/mg protein vs 53.4 ± 8.0 U/mg protein, P < .001), improved mitochondrial function, density and structure, and induced an anti-inflammatory macrophage shift, suppressing proinflammatory cytokines in vitro and in vivo. Rho-EVs showed no such effects.

CONCLUSIONS

This study showed that transecting the supraspinatus, infraspinatus tendons, and suprascapular nerve in a rat model induced muscle degeneration and fibrosis. BMSC-EVs, but not Rho-EVs, mitigated these effects by promoting an anti-inflammatory macrophage phenotype and protecting mitochondrial function through mitochondrial transfer.

CLINICAL RELEVANCE: Mitochondrial transfer via BMSC-EVs may offer a therapeutic strategy to prevent muscle degeneration in patients with rotator cuff tear.