title: Particle-Induced Osteolysis Is Mediated by TIRAP/Mal in Vitro and in Vivo: Dependence on Adherent Pathogen-Associated Molecular Patterns.
abstract: BACKGROUND
  Proinflammatory signaling by toll-like receptors (TLRs) likely contributes to biologic responses to wear particles causing aseptic loosening. We recently reported associations with aseptic loosening in patients with polymorphisms in the locus encoding an adapter protein specific for TLR-2 and TLR-4 known as toll/interleukin-1 receptor domain-containing adapter protein/MyD88 adapter-like (TIRAP/Mal). To directly examine the contribution of TIRAP/Mal, we tested the hypothesis that TIRAP/Mal deficiency reduces the activity of wear particles. Signaling by TLR-2 and TLR-4 through TIRAP/Mal can be activated by bacterial pathogen-associated molecular patterns (PAMPs) such as lipopolysaccharide or endogenous alarmins. To distinguish between those possibilities, we tested the hypothesis that the effects of TIRAP/Mal depend on the adherence of bacterial PAMPs to the particles.
  
  
  METHODS
  In vitro mRNA levels and secretion of tumor necrosis factor-α, interleukin (IL)-1β, and IL-6 were measured after incubating wild-type and TIRAP/Mal(-/-) macrophages in the presence or absence of titanium particles with adherent bacterial debris, so-called endotoxin-free particles, or particles with adherent lipopolysaccharide. In vivo osteolysis was measured after implanting titanium particles on the calvaria of wild-type and TIRAP/Mal(-/-) mice.
  
  
  RESULTS
  TIRAP/Mal deficiency significantly inhibited the activity of titanium particles with adherent bacterial debris to stimulate in vivo osteolysis and in vitro cytokine mRNAs and secretion. Those effects are dependent on adherent PAMPs because removal of >99% of the adherent bacterial debris from the particles significantly reduced their activity and the remaining activity was not dependent on TIRAP/Mal. Moreover, adherence of highly purified lipopolysaccharide to the endotoxin-free particles reconstituted the activity and the dependence on TIRAP/Mal.
  
  
  CONCLUSIONS
  TIRAP/Mal deficiency reduces inflammatory responses and osteolysis induced by particles with adherent PAMPs.
  
  
  CONCLUSIONS
  Our results, coupled with the genetic associations between aseptic loosening and polymorphisms within the TIRAP/Mal locus, support TLR signaling through TIRAP/Mal as one of the factors that enhances the activity of wear particles and further support the hypothesis that bacterial PAMPs likely contribute to aseptic loosening in a subset of patients.
authors.affiliation: Departments of Orthopaedics (C.P.B., J.J.G., J.M.T., and E.M.G.) and Pathology (E.M.G.), University Hospitals Case Medical Center, Case Western Reserve University, Cleveland, Ohio.
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Bechtel
authors.affiliation: Departments of Orthopaedics (C.P.B., J.J.G., J.M.T., and E.M.G.) and Pathology (E.M.G.), University Hospitals Case Medical Center, Case Western Reserve University, Cleveland, Ohio.
authors.forename: Jeremy J
authors.initials: JJ
authors.lastname: Gebhart
authors.affiliation: Departments of Orthopaedics (C.P.B., J.J.G., J.M.T., and E.M.G.) and Pathology (E.M.G.), University Hospitals Case Medical Center, Case Western Reserve University, Cleveland, Ohio.
authors.forename: Joscelyn M
authors.initials: JM
authors.lastname: Tatro
authors.affiliation: Departments of Cardiovascular Science (E.K.-T.) and Human Metabolism (J.M.W.), University of Sheffield, Sheffield, United Kingdom.
authors.forename: Endre
authors.initials: E
authors.lastname: Kiss-Toth
authors.affiliation: Departments of Cardiovascular Science (E.K.-T.) and Human Metabolism (J.M.W.), University of Sheffield, Sheffield, United Kingdom.
authors.forename: J Mark
authors.initials: JM
authors.lastname: Wilkinson
authors.affiliation: Departments of Orthopaedics (C.P.B., J.J.G., J.M.T., and E.M.G.) and Pathology (E.M.G.), University Hospitals Case Medical Center, Case Western Reserve University, Cleveland, Ohio emg3@cwru.edu.
authors.forename: Edward M
authors.initials: EM
authors.lastname: Greenfield
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D005260:Female; D007595:Joint Prosthesis / Q000009:adverse effects*; D008070:Lipopolysaccharides / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000172:deficiency / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010014:Osteolysis / Q000209:etiology* / Q000378:metabolism / Q000382:microbiology; D000069452:Pathogen-Associated Molecular Pattern Molecules / Q000378:metabolism*; D011475:Prosthesis Failure; D011897:Random Allocation; D017472:Receptors, Interleukin-1 / Q000172:deficiency / Q000378:metabolism*; D014025:Titanium / Q000009:adverse effects*
chemical_list: D015415:Biomarkers; D016207:Cytokines; D008070:Lipopolysaccharides; D008562:Membrane Glycoproteins; D000069452:Pathogen-Associated Molecular Pattern Molecules; D017472:Receptors, Interleukin-1; C499045:TIRAP protein, mouse; D014025:Titanium