Osteopontin deficiency impairs wear debris-induced osteolysis via regulation of cytokine secretion from murine macrophages.
OBJECTIVE
To investigate the molecular mechanisms underlying particle-induced osteolysis, we focused on osteopontin (OPN), a cytokine and cell-attachment protein that is associated with macrophage chemoattractant and osteoclast activation.


METHODS
We compared OPN protein levels in human periprosthetic osteolysis tissues with those in osteoarthritis (OA) synovial tissues. To investigate the functions of OPN during particle-induced osteolysis in vivo, titanium particles were implanted onto the calvaria of OPN-deficient mice and their wild-type (WT) littermates. Mice were killed on day 10 and evaluated immunohistologically. The effects of OPN deficiency on the secretion of inflammatory cytokines were examined using cultured bone marrow-derived macrophages (BMMs). BMMs from OPN-deficient and WT mice were cultured with titanium particles for 12 hours, and the concentrations of inflammatory cytokines in the conditioned media were measured by enzyme-linked immunosorbent assay.


RESULTS
Expression of OPN protein was enhanced in human periprosthetic osteolysis tissues as compared with OA synovial tissues. In the particle-induced model of osteolysis of the calvaria, bone resorption was significantly suppressed by OPN deficiency via inhibition of osteoclastogenesis, whereas an inflammatory reaction was observed regardless of the genotype. Results of immunostaining indicated that OPN protein was highly expressed in the membrane and bone surface at the area of bone resorption in WT mice. When BMMs were exposed to titanium particles, the concentration of proinflammatory cytokines, such as tumor necrosis factor alpha, interleukin-1alpha (IL-1alpha), IL-1beta, and IL-6, as well as chemotactic factors, such as monocyte chemoattractant protein 1 and macrophage inflammatory protein 1alpha, in the conditioned medium were significantly reduced by OPN deficiency. Whereas phagocytic activity of BMMs was not attenuated by OPN deficiency, phagocytosis-mediated NF-kappaB activation was impaired in OPN-deficient BMMs. These data indicated that OPN was implicated in the development of particle-induced osteolysis via the orchestration of pro-/antiinflammatory cytokines secreted from macrophages.


CONCLUSIONS
OPN plays critical roles in wear debris-induced osteolysis, suggesting that OPN is a candidate therapeutic target for periprosthetic osteolysis.
Shimizu
Sadanori
S
International Research Center for Molecular Science in Tooth and Bone Diseases, and Tokyo Medical and Dental University, Tokyo, Japan.
Okuda
Naoki
N
Kato
Norihiko
N
Rittling
Susan R
SR
Okawa
Atsushi
A
Shinomiya
Kenichi
K
Muneta
Takeshi
T
Denhardt
David T
DT
Noda
Masaki
M
Tsuji
Kunikazu
K
Asou
Yoshinori
Y
D000818:Animals; D002478:Cells, Cultured; D054405:Chemokine CCL3 / Q000276:immunology / Q000378:metabolism; D016207:Cytokines / Q000276:immunology / Q000378:metabolism*; D004195:Disease Models, Animal; D005260:Female; D020672:I-kappa B Proteins / Q000378:metabolism; D053582:Interleukin-1alpha / Q000276:immunology / Q000378:metabolism; D053583:Interleukin-1beta / Q000276:immunology / Q000378:metabolism; D015850:Interleukin-6 / Q000276:immunology / Q000378:metabolism; D008264:Macrophages / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D010014:Osteolysis / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D053495:Osteopontin / Q000235:genetics* / Q000378:metabolism*; D010587:Phagocytosis / Q000276:immunology; D012886:Skull / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D014025:Titanium / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology / Q000378:metabolism
C496534:Ccl3 protein, mouse; D054405:Chemokine CCL3; D016207:Cytokines; D020672:I-kappa B Proteins; D053582:Interleukin-1alpha; D053583:Interleukin-1beta; D015850:Interleukin-6; C506896:Spp1 protein, mouse; D014409:Tumor Necrosis Factor-alpha; D053495:Osteopontin; D014025:Titanium