Abstract
Mechanical loading is essential for articular cartilage homeostasis and plays a central role in the cartilage pathology, yet the mechanotransduction processes that underlie these effects remain unclear. Previously, we showed that lethal amounts of reactive oxygen species (ROS) were liberated from the mitochondria in response to mechanical insult and that chondrocyte deformation may be a source of ROS. To this end, we hypothesized that mechanically induced mitochondrial ROS is related to the magnitude of cartilage deformation. To test this, we measured axial tissue strains in cartilage explants subjected to semi-confined compressive stresses of 0, 0.05, 0.1, 0.25, 0.5, or 1.0 MPa. The presence of ROS was then determined by confocal imaging with dihydroethidium, an oxidant sensitive fluorescent probe. Our results indicated that ROS levels increased linearly relative to the magnitude of axial strains (\(r^2 = 0.87, p < 0.05\)), and significant cell death was observed at strains \(>\)40 %. By contrast, hydrostatic stress, which causes minimal tissue strain, had no significant effect. Cell-permeable superoxide dismutase mimetic Mn(III)tetrakis (1-methyl-4-pyridyl) porphyrin pentachloride significantly decreased ROS levels at 0.5 and 0.25 MPa. Electron transport chain inhibitor, rotenone, and cytoskeletal inhibitor, cytochalasin B, significantly decreased ROS levels at 0.25 MPa. Our findings strongly suggest that ROS and mitochondrial oxidants contribute to cartilage mechanobiology.
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Abbreviations
- ROS:
-
Reactive oxygen species
- DHE:
-
Dihydroethidium
- MPa:
-
Megapascal
- MnTMPyP:
-
Mn(III)tetrakis (1-methyl-4-pyridyl) porphyrin pentachloride
- EthD-2:
-
Ethidium homodimer-2
- QCIP\(^\mathrm{TM}\) :
-
Quantitative cell image processing
- Hif:
-
Hypoxia inducible factor
- tBHP:
-
Tertiary butyl hydroperoxide
- MMP-13:
-
Matrix metalloproteinase 13
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Acknowledgments
We thank Rachel Brouillette for editing. Confocal images were all taken at the Central Microscopy Research Facility, University of Iowa, Iowa City. This work was supported by the National Institutes of Health (CORT NIH P50 AR055533) and by a Merit Review Award from the Department of Veterans Affairs.
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Brouillette, M.J., Ramakrishnan, P.S., Wagner, V.M. et al. Strain-dependent oxidant release in articular cartilage originates from mitochondria. Biomech Model Mechanobiol 13, 565–572 (2014). https://doi.org/10.1007/s10237-013-0518-8
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DOI: https://doi.org/10.1007/s10237-013-0518-8