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¹ Institut National de la Santé et de la Recherche Médicale, Unité 841, Institut Mondor de Recherche Biomédicale, "Cell Interactions in the Neuromuscular System" Team, Université Paris 12 Val-de-Marne, 94000 Créteil, France
² Plateforme Cytométrie Institut Mondor de Medecine Moleculaire, IFR10, Université Paris 12 Val-de-Marne, 94000 Créteil, France
³ Vrije Universiteit, Vrije Universiteit Medisch Centrum, Department of Molecular Cell Biology, Faculty of Medicine, 1081 BT Amsterdam, Netherlands
⁴ Assistance Publique-Hôpitaux de Paris, Groupe Henri Mondor-Albert Chenevier, Service d'immunologie-Biologie, 94000 Créteil, France

CORRESPONDENCE Bénédicte Chazaud: benedicte.chazaud{at}creteil.inserm.fr

Macrophages (MPs) are important for skeletal muscle regeneration in vivo and may exert beneficial effects on myogenic cell growth through mitogenic and antiapoptotic activities in vitro. However, MPs are highly versatile and may exert various, and even opposite, functions depending on their activation state. We studied monocyte (MO)/MP phenotypes and functions during skeletal muscle repair. Selective labeling of circulating MOs by latex beads in CX3CR1^GFP/+ mice showed that injured muscle recruited only CX3CR1^lo/Ly-6C⁺ MOs from blood that exhibited a nondividing, F4/80^lo, proinflammatory profile. Then, within muscle, these cells switched their phenotype to become proliferating antiinflammatory CX3CR1^hi/Ly-6C^– cells that further differentiated into F4/80^hi MPs. In vitro, phagocytosis of muscle cell debris induced a switch of proinflammatory MPs toward an antiinflammatory phenotype releasing transforming growth factor ß1. In co-cultures, inflammatory MPs stimulated myogenic cell proliferation, whereas antiinflammatory MPs exhibited differentiating activity, assessed by both myogenin expression and fusion into myotubes. Finally, depletion of circulating MOs in CD11b–diphtheria toxin receptor mice at the time of injury totally prevented muscle regeneration, whereas depletion of intramuscular F4/80^hi MPs at later stages reduced the diameter of regenerating fibers. In conclusion, injured skeletal muscle recruits MOs exhibiting inflammatory profiles that operate phagocytosis and rapidly convert to antiinflammatory MPs that stimulate myogenesis and fiber growth.

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