A Third‐Generation Bisphosphonate, YM175, Inhibits Osteoclast Formation in Murine Cocultures by Inhibiting Proliferation of Precursor Cells via Supporting Cell‐Dependent Mechanisms

The theory that bisphosphonates inhibit osteoclast formation through their effects on osteoblastic cells remains controversial. To confirm the inhibitory effect of bisphosphonates on osteoclast formation and gain some insights into the underlying mechanisms, we examined the effect of disodium dihydrogen (cycloheptylamino)‐methylenebisphosphonate monohydrate (YM175) on osteoclast‐like multinucleated cell (OCL) formation in various mouse coculture systems. When different origins of osteoclast precursors (bone marrow, spleen, or nonspecific esterase‐positive cells) were cocultured with the same supporting cells (calvarial osteoblasts), YM175 inhibited OCL formation similarly in all cultures. When the same osteoclast precursors (spleen cells) were cocultured with supporting cells of different origin, the results were variable. YM175 inhibited OCL formation almost completely in cocultures with calvarial osteoblasts or osteoblastic cell line KS4, while it did not, or only slightly, inhibit OCL formation in cocultures with stromal cell lines, ST2 or MC3T3‐G2/PA6. Temporal addition of YM175 in cocultures of spleen cells with osteoblastic cells revealed that YM175 was effective when it was present at an early phase of the culture period. Consistent with this observation, YM175 in the presence of osteoblastic cells inhibited proliferation of preosteoclastic cells, but did not inhibit the fusion of mononuclear prefusion osteoclasts. In conclusion, the inhibitory effect of YM175 on OCL formation was confirmed in various murine coculture systems, but the effect was dependent on the types of bone‐derived cells supporting osteoclastogenesis. The findings suggest that YM175 inhibits osteoclastogenesis by inhibiting the proliferation of osteoclast precursors through its action on supporting cells of osteoblast lineage rather than acting directly on osteoclast precursors.

[1]  Roodman Gd.  Advances in bone biology: the osteoclast. , 1996, Endocrine reviews.

[2]  G. Rodan,et al.  Bisphosphonates: mechanisms of action , 1998, Endocrine reviews.

[3]  H. Fleisch,et al.  Bisphosphonates induce osteoblasts to secrete an inhibitor of osteoclast-mediated resorption. , 1996, Endocrinology.

[4]  C. Leu,et al.  Protein-tyrosine phosphatase activity regulates osteoclast formation and function: inhibition by alendronate. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[5]  A. Parfitt,et al.  Theoretical perspective: A new model for the regulation of bone resorption, with particular reference to the effects of bisphosphonates , 1996 .

[6]  G. Roodman,et al.  Advances in bone biology: the osteoclast. , 1996, Endocrine reviews.

[7]  N. Kugai,et al.  Bisphosphonates act on osteoblastic cells and inhibit osteoclast formation in mouse marrow cultures. , 1996, Bone.

[8]  T. Yoneda,et al.  Bisphosphonates promote apoptosis in murine osteoclasts in vitro and in vivo , 1995, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[9]  G. Wesolowski,et al.  Isolation and characterization of highly enriched, prefusion mouse osteoclastic cells. , 1995, Experimental cell research.

[10]  N. Udagawa,et al.  A possible mechanism of the specific action of bisphosphonates on osteoclasts: tiludronate preferentially affects polarized osteoclasts having ruffled borders. , 1995, Bone.

[11]  T. Martin,et al.  Mechanisms by which cells of the osteoblast lineage control osteoclast formation and activity , 1994, Journal of cellular biochemistry.

[12]  E. Jimi,et al.  Dexamethasone stimulates osteoclast-like cell formation by inhibiting granulocyte-macrophage colony-stimulating factor production in mouse bone marrow cultures. , 1994, Endocrinology.

[13]  C. E. Evans,et al.  Effects of two novel bisphosphonates on bone cells in vitro. , 1994, Bone and mineral.

[14]  G. Stein,et al.  Glucocorticoids promote development of the osteoblast phenotype by selectively modulating expression of cell growth and differentiation associated genes , 1992, Journal of cellular biochemistry.

[15]  H. Fleisch.  Bisphosphonates , 2012, Drugs.

[16]  G. Rodan,et al.  Bisphosphonate action. Alendronate localization in rat bone and effects on osteoclast ultrastructure. , 1991, The Journal of clinical investigation.

[17]  N. Udagawa,et al.  Role of colony‐stimulating factors in osteoclast development , 1991, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[18]  F. Bauss,et al.  BM 21.0955, a potent new bisphosphonate to inhibit bone resorption , 1991, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[19]  N. Udagawa,et al.  Cloning of an osteoblastic cell line involved in the formation of osteoclast‐like cells , 1990, Journal of cellular physiology.

[20]  T. Martin,et al.  Origin of osteoclasts: mature monocytes and macrophages are capable of differentiating into osteoclasts under a suitable microenvironment prepared by bone marrow-derived stromal cells. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[21]  T. Martin,et al.  The bone marrow-derived stromal cell lines MC3T3-G2/PA6 and ST2 support osteoclast-like cell differentiation in cocultures with mouse spleen cells. , 1989, Endocrinology.

[22]  B V Morukov,et al.  Effects of a bisphosphonate (1-hydroxy ethylidene-1,1 bisphosphonic acid) on osteoclast number during prolonged bed rest in healthy humans. , 1989, Metabolism: clinical and experimental.

[23]  Gowen,et al.  Inhibition of osteoclast-like cell formation by bisphosphonates in long-term cultures of human bone marrow. , 1989, The Journal of clinical investigation.

[24]  R J Gould,et al.  Echistatin. A potent platelet aggregation inhibitor from the venom of the viper, Echis carinatus. , 1988, The Journal of biological chemistry.

[25]  T. Martin,et al.  Osteoblastic cells are involved in osteoclast formation. , 1988, Endocrinology.

[26]  M. Ogawa,et al.  B cell ontogeny in murine embryo studied by a culture system with the monolayer of a stromal cell clone, ST2: B cell progenitor develops first in the embryonal body rather than in the yolk sac. , 1988, The EMBO journal.

[27]  Sheila J. Jones,et al.  Osteoclast-like cell formation and its regulation by osteotropic hormones in mouse bone marrow cultures. , 1988, Endocrinology.

[28]  M. Cecchini,et al.  Effect of bisphosphonates on proliferation and viability of mouse bone marrow‐derived macrophages , 1987, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[29]  P. Boonekamp,et al.  Two modes of action of bisphosphonates on osteoclastic resorption of mineralized matrix. , 1986, Bone and mineral.

[30]  H. Koyama,et al.  Hormonal responsiveness of a preadipose cell line derived from newborn mouse calvaria , 1982, Journal of cellular physiology.

[31]  K. Paigen,et al.  A simple, rapid, and sensitive DNA assay procedure. , 1980, Analytical biochemistry.