title: Real-Time H2O2Measurements in Bone Marrow Mesenchymal Stem Cells (MSCs) Show Increased Antioxidant Capacity in Cells From Osteoporotic Women.
abstract: Oxidative stress (OS) derived from an increase in intracellular reactive oxygen species (ROS) is a major determinant of aging and lifespan. It has also been associated with several age-related disorders, like postmenopausal osteoporosis of Mesenchymal stem cells (MSCs). MSCs are the common precursors for osteoblasts and adipocytes; appropriate commitment and differentiation of MSCs into a specific phenotype is modulated, among other factors, by ROS balance. MSCs have shown more resistance to ROS than differentiated cells, and their redox status depends on complex and abundant anti-oxidant mechanisms. The purpose of this work was to analyze in real time, H2O2signaling in individual h-MSCs, and to compare the kinetic parameters of H2O2management by cells derived from both control (c-) and osteoporotic (o-) women. For these purposes, cells were infected with a genetically encoded fluorescent biosensor named HyPer, which is specific for detecting H2O2inside living cells. Subsequently, cells were sequentially challenged with 50 and 500 μM H2O2pulses, and the cellular response was recorded in real time. The results demonstrated adequate expression of the biosensor allowing registering fluorescence from HyPer at a single cell level. Comparison of the response of c- and o-MSCs to the oxidant challenges demonstrated improved antioxidant activity in o-MSCs. This was further corroborated by measuring the relative expression of mRNAs for catalase, superoxide dismutase-1, thioredoxine, and peroxiredoxine, as well as by cell-surviving capacity under short-term H2O2treatment. We conclude that functional differences exist between healthy and osteoporotic human MSCs. The mechanism for these differences requires further study. J. Cell. Biochem. 118: 585-593, 2017. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Laboratorio de Biología Celular, Instituto de Nutrición y Tecnología de los Alimentos, Universidad de Chile, Santiago, Chile.
authors.forename: Flavia
authors.initials: F
authors.lastname: Román
authors.affiliation: Laboratorio de Biología Celular, Instituto de Nutrición y Tecnología de los Alimentos, Universidad de Chile, Santiago, Chile.
authors.forename: Carla
authors.initials: C
authors.lastname: Urra
authors.affiliation: Laboratorio de Biología Celular, Instituto de Nutrición y Tecnología de los Alimentos, Universidad de Chile, Santiago, Chile.
authors.forename: Omar
authors.initials: O
authors.lastname: Porras
authors.affiliation: Laboratorio de Biología Celular, Instituto de Nutrición y Tecnología de los Alimentos, Universidad de Chile, Santiago, Chile.
authors.forename: Ana María
authors.initials: AM
authors.lastname: Pino
authors.affiliation: Maine Medical Center Research Institute, Scarborough, Maine.
authors.forename: Clifford J
authors.initials: CJ
authors.lastname: Rosen
authors.affiliation: Laboratorio de Biología Celular, Instituto de Nutrición y Tecnología de los Alimentos, Universidad de Chile, Santiago, Chile.
authors.forename: Juan Pablo
authors.initials: JP
authors.lastname: Rodríguez
mesh_terms: D000368:Aged; D000975:Antioxidants / Q000378:metabolism*; D001854:Bone Marrow Cells / Q000378:metabolism* / Q000473:pathology; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism*; D059630:Mesenchymal Stem Cells / Q000378:metabolism* / Q000473:pathology; D008875:Middle Aged; D015663:Osteoporosis, Postmenopausal / Q000378:metabolism* / Q000473:pathology; D010088:Oxidoreductases / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D000975:Antioxidants; D006861:Hydrogen Peroxide; D010088:Oxidoreductases
keywords: ANTIOXIDANT CAPACITY; MESENCHYMAL STEM CELLS; OSTEOPOROSIS