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Biological Research

Print version ISSN 0716-9760

Abstract

MURILLO, Andrea; GUERRERO, Carlos A; ACOSTA, Orlando  and  CARDOZO, Carmen A. Bone resorptive activity of osteoclast-like cells generated in vitro by PEG-induced macrophage fusion. Biol. Res. [online]. 2010, vol.43, n.2, pp.205-224. ISSN 0716-9760.  http://dx.doi.org/10.4067/S0716-97602010000200009.

Normal bone remodeling is maintained by a balance between osteoclast and osteoblast activity, whereas defects in osteoclast activity affecting such balance result in metabolic bone disease. Macrophage-macrophage fusion leading to multinucleated osteoclasts being formed is still not well understood. Here we present PEG-induced fusion of macrophages from both U937/A and J774 cell lines and the induced differentiation and activation of osteoclast-like cells according to the expression of osteoclast markers such as tartrate resistant acid phosphatase (TRAP) and bone resorptive activity. PEG-induced macrophage fusion, during the non-confuent stage, signifcantly increased the osteoclastogenic activity of macrophages from cell lines compared to that of spontaneous cell fusion in the absence of PEG (polyethylene glycol). The results shown in this work provide evidence that cell fusion per se induces osteoclast-like activity. PEG-fused macrophage differential response to pretreatment with osteoclastogenic factors was also examined in terms of its ability to form TRAP positive multinucleated cells (TPMNC) and its resorptive activity on bovine cortical bone slices. Our work has also led to a relatively simple method regarding those previously reported involving cell co-cultures. Multinucleated osteoclast-like cells obtained by PEG-induced fusion of macrophages from cell lines could represent a suitable system for conducting biochemical studies related to basic macrophage fusion mechanisms, bone-resorption activity and the experimental search for bone disease therapeutic alternatives.

Keywords : bone resorption; macrophage fusion; osteoclast activity; osteoclast differentiation; PEG; RANKL.

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