title: Regulation of RANKL by biomechanical loading in fibrochondrocytes of meniscus.
abstract: OBJECTIVE
  We sought to determine whether fibrochondrocytes from menisci express receptor activator of NF-kappaB (RANK), its ligand (RANKL), or osteoprotegerin (OPG) and, if so, whether their expression is modulated by dynamic mechanical loading under inflammatory and normal conditions.
  
  
  METHODS
  Fibrochondrocytes from rat menisci were subjected to cyclic tensile strain (CTS) at various magnitudes and frequencies in the presence or absence of interleukin (IL)-1beta for up to 24 h. In order to determine whether a possible regulatory effect of mechanical loading on RANKL and its receptors under inflamed conditions is sustained, cells were stimulated with IL-1beta for 24 h while being subjected to CTS only for the initial 4 and 8h, respectively. Regulation of RANKL, RANK, and OPG expression and synthesis were determined by semiquantitative and real-time PCR, Western blotting, and immunofluorescence.
  
  
  RESULTS
  Fibrochondrocytes constitutively expressed low levels of RANKL and RANK but marked levels of OPG. IL-1beta upregulated expression and synthesis of RANKL and RANK significantly (p<0.05), whereas expression of OPG was unaffected following 4 and 24 h. When fibrochondrocytes were simultaneously subjected to CTS and IL-1beta, expression of RANKL and RANK was significantly (p<0.05) downregulated as compared to that of IL-1beta-stimulated unstretched cells. The inhibitory effect of CTS on the IL-1beta-induced upregulation of RANKL and RANK was sustained as well as magnitude and frequency dependent.
  
  
  CONCLUSIONS
  Our study provides evidence that RANKL and its receptors are expressed in fibrochondrocytes from meniscus. These data also demonstrate that dynamic mechanical loading can modify the expression of RANKL and RANK in inflammatory conditions.
authors.affiliation: Biomechanics and Tissue Engineering Laboratory, 4010 Postle Hall, College of Dentistry, The Ohio State University, 305 W 12th Avenue, Columbus, 43210, USA.
authors.forename: James
authors.initials: J
authors.lastname: Deschner
authors.forename: Ewa
authors.initials: E
authors.lastname: Wypasek
authors.forename: Mario
authors.initials: M
authors.lastname: Ferretti
authors.forename: Birgit
authors.initials: B
authors.lastname: Rath
authors.forename: Mirela
authors.initials: M
authors.lastname: Anghelina
authors.forename: Sudha
authors.initials: S
authors.lastname: Agarwal
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D007249:Inflammation / Q000378:metabolism; D053583:Interleukin-1beta / Q000494:pharmacology; D008592:Menisci, Tibial / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D053244:Osteoprotegerin / Q000235:genetics / Q000378:metabolism; D053245:RANK Ligand / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D053246:Receptor Activator of Nuclear Factor-kappa B / Q000235:genetics / Q000378:metabolism; D013314:Stress, Mechanical
chemical_list: D053583:Interleukin-1beta; D053244:Osteoprotegerin; D053245:RANK Ligand; D012333:RNA, Messenger; D053246:Receptor Activator of Nuclear Factor-kappa B