title: Radiation-induced alterations of osteogenic and chondrogenic differentiation of human mesenchymal stem cells.
abstract: While human mesenchymal stem cells (hMSCs), either in the bone marrow or in tumour microenvironment could be targeted by radiotherapy, their response is poorly understood. The oxic effects on radiosensitivity, cell cycle progression are largely unknown, and the radiation effects on hMSCs differentiation capacities remained unexplored. Here we analysed hMSCs viability and cell cycle progression in 21% O2 and 3% O2 conditions after medical X-rays irradiation. Differentiation towards osteogenesis and chondrogenesis after irradiation was evaluated through an analysis of differentiation specific genes. Finally, a 3D culture model in hypoxia was used to evaluate chondrogenesis in conditions mimicking the natural hMSCs microenvironment. The hMSCs radiosensitivity was not affected by O2 tension. A decreased number of cells in S phase and an increase in G2/M were observed in both O2 tensions after 16 hours but hMSCs released from the G2/M arrest and proliferated at day 7. Osteogenesis was increased after irradiation with an enhancement of mRNA expression of specific osteogenic genes (alkaline phosphatase, osteopontin). Osteoblastic differentiation was altered since matrix deposition was impaired with a decreased expression of collagen I, probably through an increase of its degradation by MMP-3. After induction in monolayers, chondrogenesis was altered after irradiation with an increase in COL1A1 and a decrease in both SOX9 and ACAN mRNA expression. After induction in a 3D culture in hypoxia, chondrogenesis was altered after irradiation with a decrease in COL2A1, ACAN and SOX9 mRNA amounts associated with a RUNX2 increase. Together with collagens I and II proteins decrease, associated to a MMP-13 expression increase, these data show a radiation-induced impairment of chondrogenesis. Finally, a radiation-induced impairment of both osteogenesis and chondrogenesis was characterised by a matrix composition alteration, through inhibition of synthesis and/or increased degradation. Alteration of osteogenesis and chondrogenesis in hMSCs could potentially explain bone/joints defects observed after radiotherapy.
authors.affiliation: Normandy University, Caen, France; UNICAEN, Laboratoire Microenvironnement Cellulaire et Pathologies (MILPAT), Caen, France; Laboratoire Accueil en Radiobiologie avec les Ions Accélérés (CEA-DSV-IRCM-LARIA), Bd Becquerel, Caen Cedex 5, France; Regenerative Medicine Institute (REMEDI), National University of Ireland Galway, Galway, Ireland.
authors.forename: Séverine
authors.initials: S
authors.lastname: Cruet-Hennequart
authors.affiliation: Normandy University, Caen, France; UNICAEN, Laboratoire Microenvironnement Cellulaire et Pathologies (MILPAT), Caen, France.
authors.forename: Carole
authors.initials: C
authors.lastname: Drougard
authors.affiliation: Regenerative Medicine Institute (REMEDI), National University of Ireland Galway, Galway, Ireland.
authors.forename: Georgina
authors.initials: G
authors.lastname: Shaw
authors.affiliation: Normandy University, Caen, France; UNICAEN, Laboratoire Microenvironnement Cellulaire et Pathologies (MILPAT), Caen, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Legendre
authors.affiliation: Normandy University, Caen, France; UNICAEN, Laboratoire Microenvironnement Cellulaire et Pathologies (MILPAT), Caen, France.
authors.forename: Magali
authors.initials: M
authors.lastname: Demoor
authors.affiliation: Regenerative Medicine Institute (REMEDI), National University of Ireland Galway, Galway, Ireland.
authors.forename: Frank
authors.initials: F
authors.lastname: Barry
authors.affiliation: Laboratoire Accueil en Radiobiologie avec les Ions Accélérés (CEA-DSV-IRCM-LARIA), Bd Becquerel, Caen Cedex 5, France.
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Lefaix
authors.affiliation: Normandy University, Caen, France; UNICAEN, Laboratoire Microenvironnement Cellulaire et Pathologies (MILPAT), Caen, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Galéra
mesh_terms: D000293:Adolescent; D000328:Adult; D002453:Cell Cycle / Q000528:radiation effects; D002454:Cell Differentiation / Q000528:radiation effects*; D002460:Cell Line; D049109:Cell Proliferation / Q000528:radiation effects; D002470:Cell Survival / Q000528:radiation effects; D016922:Cellular Senescence / Q000528:radiation effects; D020219:Chondrogenesis; D003094:Collagen / Q000235:genetics / Q000378:metabolism; D059565:G2 Phase Cell Cycle Checkpoints / Q000528:radiation effects; D015870:Gene Expression; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000528:radiation effects*; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism / Q000528:radiation effects; D010012:Osteogenesis; D010101:Oxygen Consumption; D014965:X-Rays; D055815:Young Adult
chemical_list: D003094:Collagen
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