Bioenergetic reprogramming of articular chondrocytes by exposure to exogenous and endogenous reactive oxygen species and its role in the anabolic response to low oxygen

Journal of Tissue Engineering and Regenerative Medicine
H K Heywood, D A Lee

Abstract

Monolayer culture is integral to many cell-based cartilage repair strategies, but chondrocytes lose regenerative potential with increasing duration in vitro. This coincides with elevated reactive oxygen species (ROS) levels and a bioenergetic transformation characterized by increasing mitochondrial function. This study investigates ROS as stimuli for bioenergetic reprogramming and the effect of antioxidants on the propensity of chondrocytes to regenerate a cartilaginous matrix. Articular chondrocytes were cultured in monolayer under a 2% O2 atmosphere. Oxidative stress was increased using 50 μm H2 O2 or a 20% O2 culture atmosphere, or decreased using the antioxidant N-acetyl-cysteine (NAC). Mitochondrial function was characterized using 200 nm Mitotracker green and an oxygen biosensor. After two population doublings ± NAC, chondrocytes were encapsulated in alginate beads (1 × 107 cells/ml) for an additional 10 days before DMB assay of glycosaminoglycan content. The beads were cultured under both 20% O2 and the more physiological 5% O2 condition. Chondrocytes expanded in 20% O2 exhibited elevated mitochondrial mass and functional capacity, which was partially mimicked by the exogenous ROS, H2 O2 . Oligomycin treatment revealed t...Continue Reading

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Citations

Jul 20, 2016·Oxidative Medicine and Cellular Longevity·Jiyuan BuGang Chen
Jul 21, 2017·Analytical Chemistry·Brenda BakkerBerta Cillero-Pastor
Oct 26, 2018·Physiological Reviews·Thomas P Keeley, Giovanni E Mann
Mar 8, 2021·Free Radical Biology & Medicine·Ryan A Denu, Peiman Hematti

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Methods Mentioned

BETA
light microscopy
biosensor
PCR

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