title: Hydrostatic Pressure Regulates MicroRNA Expression Levels in Osteoarthritic Chondrocyte Cultures via the Wnt/β-Catenin Pathway.
abstract: Mechanical loading and hydrostatic pressure (HP) regulate chondrocytes' metabolism; however, how mechanical stimulation acts remain unclear. MicroRNAs (miRNAs) play an important role in cartilage homeostasis, mechanotransduction, and in the pathogenesis of osteoarthritis (OA). This study investigated the effects of a cyclic HP (1-5 MPa), in both normal and OA human chondrocytes, on the expression ofmiR-27a/b,miR-140,miR-146a/b, andmiR-365, and of their target genes (MMP-13,ADAMTS-5,IGFBP-5, andHDAC-4). Furthermore, we assessed the possible involvement of Wnt/β-catenin pathway in response to HP. Chondrocytes were exposed to HP for 3h and the evaluations were performed immediately after pressurization, and following 12, 24, and 48 h. Total RNA was extracted and used for real-time PCR. β-catenin was detected by Western blotting analysis and immunofluorescence. In OA chondrocytes, HP induced a significant increase (p< 0.01) of the expression levels ofmiR-27a/b,miR-140, andmiR-146a, and a significant reduction (p< 0.01) ofmiR-365at all analyzed time points.MMP-13,ADAMTS-5, andHDAC-4were significantly downregulated following HP, while no significant modification was found forIGFBP-5. β-catenin levels were significantly increased (p< 0.001) in OA chondrocytes at basal conditions and significantly reduced (p< 0.01) by HP. Pressurization did not cause any significant modification in normal cells. In conclusion, in OA chondrocytes, HP restores the expression levels of some miRNAs, downregulates MMP-13, ADAMTS-5, and HDAC-4, and modulates the Wnt/β-catenin pathway activation.
authors.affiliation: Department of Medicine, Surgery and Neuroscience, Rheumatology Unit, University of Siena, Policlinico Le Scotte, Viale Bracci 1, 53100 Siena, Italy. saracheleschi@hotmail.com.|Department of Medical Biotechnologies, University of Siena, Policlinico Le Scotte, Viale Bracci 1, 53100 Siena, Italy. saracheleschi@hotmail.com.
authors.forename: Sara
authors.initials: S
authors.lastname: Cheleschi
authors.affiliation: Department of Medicine, Surgery and Neuroscience, Rheumatology Unit, University of Siena, Policlinico Le Scotte, Viale Bracci 1, 53100 Siena, Italy. annadepalma90@live.it.|Department of Medical Biotechnologies, University of Siena, Policlinico Le Scotte, Viale Bracci 1, 53100 Siena, Italy. annadepalma90@live.it.
authors.forename: Anna
authors.initials: A
authors.lastname: De Palma
authors.affiliation: Department of Life Sciences and Biotechnology, University of Ferrara, Via Borsari, 44121 Ferrara, Italy. ale.pecorelli@gmail.com.|Child Neuropsychiatry Unit, University Hospital, AOUS, Viale Bracci, 53100 Siena, Italy. ale.pecorelli@gmail.com.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Pecorelli
authors.affiliation: Department of Medicine, Surgery and Neuroscience, Rheumatology Unit, University of Siena, Policlinico Le Scotte, Viale Bracci 1, 53100 Siena, Italy. pascarelli@unisi.it.
authors.forename: Nicola Antonio
authors.initials: NA
authors.lastname: Pascarelli
authors.affiliation: Department of Life Sciences and Biotechnology, University of Ferrara, Via Borsari, 44121 Ferrara, Italy. giuseppe.valacchi@unife.it.|Plants for Human Health Institute, Department of Animal Sciences, NC State University, NC Research Campus, 600 Laureate Way, Kannapolis, NC 2808, USA. giuseppe.valacchi@unife.it.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Valacchi
authors.affiliation: Department of Life Sciences and Biotechnology, University of Ferrara, Via Borsari, 44121 Ferrara, Italy. belmonte.giuseppe@tiscali.it.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Belmonte
authors.affiliation: Department of Medicine, Surgery and Neurosciences, Orthopaedic Section, University of Siena, Policlinico Le Scotte, Viale Bracci 1, 53100 Siena, Italy. s.carta@ao-siena.toscana.it.
authors.forename: Serafino
authors.initials: S
authors.lastname: Carta
authors.affiliation: Department of Medicine, Surgery and Neuroscience, Rheumatology Unit, University of Siena, Policlinico Le Scotte, Viale Bracci 1, 53100 Siena, Italy. mauro.galeazzi@unisi.it.
authors.forename: Mauro
authors.initials: M
authors.lastname: Galeazzi
authors.affiliation: Department of Medicine, Surgery and Neuroscience, Rheumatology Unit, University of Siena, Policlinico Le Scotte, Viale Bracci 1, 53100 Siena, Italy. fioravanti7@virgilio.it.
authors.forename: Antonella
authors.initials: A
authors.lastname: Fioravanti
mesh_terms: D000071116:ADAMTS5 Protein / Q000235:genetics; D000368:Aged; D015153:Blotting, Western; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation; D006655:Histone Deacetylases / Q000235:genetics; D006801:Humans; D006874:Hydrostatic Pressure; D018975:Insulin-Like Growth Factor Binding Protein 5 / Q000235:genetics; D008297:Male; D053509:Matrix Metalloproteinase 13 / Q000235:genetics; D035683:MicroRNAs / Q000235:genetics*; D008875:Middle Aged; D010003:Osteoarthritis / Q000473:pathology; D012097:Repressor Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D060449:Wnt Signaling Pathway; D051176:beta Catenin / Q000378:metabolism*
chemical_list: D018975:Insulin-Like Growth Factor Binding Protein 5; C542542:MIRN146 microRNA, human; C531388:MIRN27 microRNA, human; C546819:MIRN365 microRNA, human; D035683:MicroRNAs; C544302:Mirn140 microRNA, human; D012097:Repressor Proteins; D051176:beta Catenin; D000071116:ADAMTS5 Protein; C121532:ADAMTS5 protein, human; D053509:Matrix Metalloproteinase 13; C400300:HDAC4 protein, human; D006655:Histone Deacetylases
keywords: Wnt/β-catenin; chondrocyte; hydrostatic pressure; mechanical loading; microRNA; osteoarthritis