title: Platelet-rich plasma stimulates cytokine expression and alkaline phosphatase activity in osteoblast-derived osteosarcoma cells.
abstract: OBJECTIVE
  The aim of this study was to investigate the effects of PRP on SAOS-2 cells in terms of cytokine expression, cell activity and oxidative stress.
  
  
  METHODS
  Cell line SAOS-2 (1×10(5)cells/mL) were grown in culture medium α-MEM with 10% FBS for 24h and stimulated (or not) with PRP at concentrations of 3, 10 and 20%, LPS (E. coli, 10g/mL) and IL-1β (1mg/mL) for 24h. The supernatant was collected and analyzed for the expression of cytokines in a panel array, ALP using a commercial kit and NO(2)(-) with Griess reaction method. Also, the cells were analyzed using Western blot for RANKL and slot blotting for nitrotyrosine expression.
  
  
  RESULTS
  There were no significant differences amongst the groups in terms of NO(2)(-), protein nitrotyrosine content and RANKL expression. However, all stimuli increased ALP activity and in case of PRP, it was in a dose-dependent manner (p<0.001). Also, all stimuli induced an increase in cytokines and chemokines expression, but only PRP promoted an increase of component C5, sICAM-1 and RANTES expression. Whilst IL-1 receptor antagonist (IL-1ra) expression was down-regulated by PRP, both LPS and IL-1β caused up-regulation of this cytokine.
  
  
  CONCLUSIONS
  PRP can stimulate osteoblast activity and cytokine/chemokine release, as well as indicate some of the mediators that can (and cannot) be involved in this activation.
authors.affiliation: Department of Physiology and Pathology, FOAr, University of São Paulo, Araraquara, SP, Brazil. brunoherreras@gmail.com
authors.forename: Bruno S
authors.initials: BS
authors.lastname: Herrera
authors.forename: Leila S
authors.initials: LS
authors.lastname: Coimbra
authors.forename: Alliny S
authors.initials: AS
authors.lastname: Bastos
authors.forename: Simone A
authors.initials: SA
authors.lastname: Teixeira
authors.forename: Joao P
authors.initials: JP
authors.lastname: Steffens
authors.forename: Marcelo N
authors.initials: MN
authors.lastname: Muscara
authors.forename: Luis C
authors.initials: LC
authors.lastname: Spolidorio
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis*; D045744:Cell Line, Tumor; D018946:Chemokine CCL5 / Q000032:analysis; D054360:Chemokine CXCL1 / Q000032:analysis; D003182:Complement C5 / Q000032:analysis; D016207:Cytokines / Q000032:analysis*; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000032:analysis; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000032:analysis; D053590:Interleukin 1 Receptor Antagonist Protein / Q000032:analysis; D053583:Interleukin-1beta / Q000494:pharmacology; D007378:Interleukins / Q000032:analysis; D008070:Lipopolysaccharides / Q000494:pharmacology; D009569:Nitric Oxide / Q000032:analysis; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism*; D018384:Oxidative Stress / Q000502:physiology; D053657:Platelet-Rich Plasma / Q000502:physiology*; D053245:RANK Ligand / Q000032:analysis; D014443:Tyrosine / Q000031:analogs & derivatives / Q000032:analysis
chemical_list: C483024:CCL5 protein, human; D018946:Chemokine CCL5; D054360:Chemokine CXCL1; D003182:Complement C5; D016207:Cytokines; C507763:IL1RN protein, human; C496971:IL32 protein, human; D053590:Interleukin 1 Receptor Antagonist Protein; D053583:Interleukin-1beta; D007378:Interleukins; D008070:Lipopolysaccharides; D053245:RANK Ligand; C505688:TNFSF11 protein, human; D018799:Intercellular Adhesion Molecule-1; D009569:Nitric Oxide; C002744:3-nitrotyrosine; D014443:Tyrosine; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D000469:Alkaline Phosphatase