Thrombopoietin inhibits in vitro osteoclastogenesis from murine bone marrow cells.
To determine whether thrombopoietin (TPO) can modulate the osteoclastic differentiation from hematopoietic stem cells, we investigated the effect of TPO on in vitro osteoclastogenesis by using the coculture of murine bone marrow cells with the stromal cell line (ST2) in the presence of 1alpha,25-dihydroxyvitamin D3 and dexamethasone. Recombinant human TPO inhibited the formation of tartrate-resistant acid phosphatase-positive multinucleated cells in a dose-dependent manner (0.02-200 ng/ml). The effect of TPO on differentiation of bone-resorbing capacity was investigated by pit assay. TPO dose dependently decreased the areas oftoluidine blue-stained resorption pits (2.0-200 ng/ml). To identify the cellular target of TPO, we used a variety of bone marrow/stromal cell coculture methods. Initially, we found that TPO mainly exerted its effect on the early stage of osteoclastic differentiation in delayed addition experiments. Consequently, the majority of TPO's inhibition of osteoclastic cell formation was due to its effect on bone marrow cells. Finally, we examined whether transforming growth factor-beta (TGFbeta) and platelet-derived growth factor (PDGF), major cytokines produced by megakaryocytes, mediate the inhibitory effect of TPO. The addition of either anti-TGFbeta or anti-PDGF antibody to bone marrow cell culture completely antagonized the effect of TPO on osteoclastogenesis. Furthermore, treatment of bone marrow cells with TGFbeta or PDGF mimicked the inhibitory effect of TPO. These data suggest that TPO inhibits osteoclastogenesis through stimulating thrombopoiesis and that TGFbeta and PDGF mediate the effect of TPO by impacting on macrophage-lineage cells as osteoclast precursors.
Wakikawa
T
T
Department of Biochemistry, Osaka City University Medical School, Osaka, Japan.
Shioi
A
A
Hino
M
M
Inaba
M
M
Nishizawa
Y
Y
Tatsumi
N
N
Morii
H
H
Otani
S
S
D000704:Analysis of Variance; D000818:Animals; D001853:Bone Marrow / Q000187:drug effects; D001854:Bone Marrow Cells; D002117:Calcitriol / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002478:Cells, Cultured; D018920:Coculture Techniques; D016207:Cytokines / Q000378:metabolism; D003907:Dexamethasone / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006651:Histocytochemistry; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008264:Macrophages / Q000166:cytology / Q000187:drug effects; D008533:Megakaryocytes / Q000187:drug effects; D051379:Mice; D008809:Mice, Inbred C3H; D010010:Osteoclasts / Q000166:cytology* / Q000187:drug effects; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects; D013926:Thrombopoietin / Q000494:pharmacology*; D016211:Transforming Growth Factor alpha / Q000494:pharmacology
D016207:Cytokines; D010982:Platelet-Derived Growth Factor; D016211:Transforming Growth Factor alpha; D003907:Dexamethasone; D013926:Thrombopoietin; D002117:Calcitriol