title: Bone resorptive activity of human peripheral blood mononuclear cells after fusion with polyethylene glycol.
abstract: The bone remodeling process occurs through bone formation by osteoblasts and bone resorption by osteoclasts, a process involving the contribution of endocrine and nervous systems. The mechanisms associated to differentiation and proliferation of osteoclasts and osteoblasts are considered a potential therapeutic target for treating some erosive bone diseases. The aim of the present study is to explore the feasibility of generating active osteoclast-like cells from peripheral blood mononuclear cells (PBMCs) following polyethylene glycol (PEG)-induced fusion. PEG-fused PBMCs showed TRAP+-multinucleated cells and bone resorption activity, and were also positive for osteoclast markers such as carbonic anhydrase II, calcitonin receptor, vacuolar ATPase, and cathepsin K, when examined by reverse transcription-polymerase chain reaction, immunochemistry and Western blotting. TRAP expression and bone resorptive activity were higher in whole PEG-fused PBMCs than in separated T lymphocytes, B lymphocytes or monocytes. Both TRAP expression and bone resorptive activity were also higher in osteogenesis imperfecta patients compared to PEG-fused PBMCs from healthy individuals. PEG-induced fusion was more efficient in inducing TRAP and bone resorptive activities than macrophage colony-stimulating factor or dexamethasone treatment. Bone resorptive activity of PEG-fused PMBCs was inhibited by bisphosphonates. Evidence is provided that the use of PEG-based cell fusion is a straightforward and amenable method for studying human osteoclast differentiation and testing new therapeutic strategies.
authors.affiliation: Departamento de Ciencias Fisiológicas, Facultad de Medicina, Universidad Nacional de Colombia, Carrera 30 No. 45-03, Bogotá, Colombia.
authors.forename: Edwin
authors.initials: E
authors.lastname: Manrique
authors.affiliation: Departamento de Ciencias Fisiológicas, Facultad de Medicina, Universidad Nacional de Colombia, Carrera 30 No. 45-03, Bogotá, Colombia.
authors.forename: Luz M
authors.initials: LM
authors.lastname: Castillo
authors.affiliation: Departamento de Ortopedia, Facultad de Medicina, Universidad Nacional de Colombia, Bogotá, Colombia.
authors.forename: Oswaldo
authors.initials: O
authors.lastname: Lazala
authors.affiliation: Departamento de Ciencias Fisiológicas, Facultad de Medicina, Universidad Nacional de Colombia, Carrera 30 No. 45-03, Bogotá, Colombia. caguerrerof@unal.edu.co.
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Guerrero
authors.affiliation: Departamento de Ciencias Fisiológicas, Facultad de Medicina, Universidad Nacional de Colombia, Carrera 30 No. 45-03, Bogotá, Colombia.
authors.forename: Orlando
authors.initials: O
authors.lastname: Acosta
mesh_terms: D000293:Adolescent; D001402:B-Lymphocytes / Q000166:cytology / Q000378:metabolism; D001862:Bone Resorption / Q000378:metabolism*; D018929:Cell Culture Techniques; D002459:Cell Fusion; D002478:Cells, Cultured; D002648:Child; D003907:Dexamethasone / Q000494:pharmacology; D004164:Diphosphonates / Q000494:pharmacology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000378:metabolism; D016173:Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D009000:Monocytes / Q000166:cytology / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism; D010010:Osteoclasts / Q000166:cytology* / Q000378:metabolism; D010013:Osteogenesis Imperfecta / Q000503:physiopathology; D011092:Polyethylene Glycols / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism; D000071681:Tartrate-Resistant Acid Phosphatase / Q000378:metabolism
chemical_list: D004164:Diphosphonates; D011092:Polyethylene Glycols; D003907:Dexamethasone; D016173:Macrophage Colony-Stimulating Factor; C000605510:ACP5 protein, human; D000071681:Tartrate-Resistant Acid Phosphatase
keywords: Bone resorption; Monocyte fusion; Osteoclast activity; Peripheral blood mononuclear cells; Polyethylene glycol