Abstract
Background aims
Skeletal muscle regeneration after severe damage is reliant on local stem cell proliferation
and differentiation, processes that are tightly regulated by macrophages. Peripheral
artery disease is a globally prevalent cardiovascular disease affecting millions of
people. Progression of the disease leads to intermittent claudication, subsequent
critical limb ischemia and muscle injury. Tissue-derived and ex vivo–expanded mesenchymal stromal cells (MSCs) for skeletal muscle regeneration have been
studied, but pre-clinical and clinical results have not been consistent. As a result,
the potential therapeutic efficacy and associated repair mechanisms of MSCs remain
unclear. Numerous studies have demonstrated the vulnerability of delivered MSCs, with
a precipitous drop in cell viability upon transplantation. This has prompted investigation
into the therapeutic benefit of apoptotic cells, microvesicles, exosomes and soluble
signals that are released upon cell death.
Methods
In this study, we characterized various components produced by MSCs after cell death
induction under different conditions. We discovered anti-inflammatory and pro-regenerative
effects produced by cell components following a freeze and thaw (F&T) process on macrophage
polarization in vitro. We further investigated the underlying mechanisms of macrophage polarization by
those components resulting from severe cell death induction.
Results
We found potent therapeutic effects from F&T-induced cell debris are dependent on
the externalization of phosphatidylserine on the plasma membrane. In contrast, effects
from the supernatant of F&T-induced cell death primarily depends on the released protein
content. We then applied the F&T-induced cell supernatant to an animal model of peripheral
artery disease to treat muscle injury caused by severe ischemia. Treatment with the
F&T supernatant but not the vulnerable MSCs resulted in significantly improved recovery
of muscle function, blood flow and morphology and inflammation resolution in the affected
muscles 2 weeks after injury.
Conclusions
This study validates the therapeutic potential of F&T-induced supernatant obviating
the need for a viable population from vulnerable MSCs to treat injury, thus providing
a roadmap for cell-free therapeutic approaches for tissue regeneration.
Graphical Abstract
Graphical Abstract
Key Words
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Article info
Publication history
Published online: December 20, 2022
Accepted:
November 28,
2022
Received:
August 3,
2022
Publication stage
In Press Corrected ProofIdentification
Copyright
© 2022 International Society for Cell & Gene Therapy. Published by Elsevier Inc. All rights reserved.
