Abstract
Objectives: Several in vitro models have been established to investigate the effects of mechanical stimuli on osteoclasts (OCs). Despite these models, the mechanisms by which mechanical forces trigger responses and the nature of such responses are still poorly understood. Generation of reactive oxygen species (ROS) levels and their relationship with bone resorption activity of OCs under the influence of these mechanical forces have not yet been reported.
Methods: Centrifugal force was applied to human macrophage cell line U937, peripheral blood mononuclear cells (PBMCs) or periodontal ligament (PDL) fibroblasts that had been treated or not with polyethylene glycol (PEG) or N-acetylcysteine (NAC). Osteoclast markers such as tartrate-resistant acid phosphatase (TRAP) and bone resorption activities were measured. ROS levels and actin ring formation were also determined. Responses of U937 cells to centrifugal force were compared to those of PEG-induced cell fusion.
Results: The present findings show that individual cells subjected to centrifugal force stress responded by increasing intracellular levels of ROS, forming actin-like rings, expressing TRAP and bone resorption activities, and expressing typical osteoclast markers. All these responses were also detected in PEG-fused U937 cells used as a control. All the above responses were decreased by treating the cells with NAC.
Conclusion: Centrifugal force and/or PEG-induced cell fusion can induce osteoclast-like activities, including oxidative stress. The present experimental model allows us to hypothesize mechanisms underlying bone resorption involved in pathological
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