Bisphosphonates induce apoptosis in mouse macrophage-like cells in vitro by a nitric oxide-independent mechanism

Michael John Rogers, K M Chilton, Fraser Coxon, J Lawry, M. Olivia Smith, S Suri, R. Graham G. Russell

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Abstract

Bisphosphonates (BPs) are an important class of antiresorptive drugs used in the treatment of bone diseases, including osteoporosis. Although their mechanism of action has not been identified at the molecular level, there is substantial evidence that BPs can have a direct effect on osteoclasts by mechanisms that may lead to osteoclast cell death by apoptosis. BPs can also inhibit proliferation and cause cell death in macrophages in vitro. We have now shown that the toxic effect of BPs on macrophages is also due to the induction of apoptotic, rather than necrotic, cell death. Morphological and biochemical features that are definitive of apoptosis (chromatin condensation, nuclear fragmentation, and endonuclease-mediated internucleosomal cleavage of DNA) could be identified in mouse macrophage-like J774 and RAW264 cells, following treatment with 100 microM pamidronate, alendronate, and ibandronate for 24 h or more. Clodronate was much less potent, even at 2000 microM, while 2000 microM etidronate did not cause apoptosis. Apoptosis was not due to increased synthesis of nitric oxide and could not be prevented by inhibitors of nitric oxide synthases. Since macrophages, like osteoclasts, are particularly susceptible to BPs, these observations support the recent suggestion that the mechanism by which BPs inhibit bone resorption may involve osteoclast apoptosis. Furthermore, the macrophage-like cell lines used in this study may be a convenient model with which to identify the molecular mechanisms by which BPs promote apoptosis in osteoclasts. Induction of macrophage apoptosis by BPs in vivo may also account, at least in part, for the anti-inflammatory properties of BPs as well as the ability of BPs to cause an acute phase response.
Original languageEnglish
Pages (from-to)1482-1491
Number of pages10
JournalJournal of Bone and Mineral Research
Volume11
Issue number10
DOIs
Publication statusPublished - 1 Oct 1996

Fingerprint

Diphosphonates
Nitric Oxide
Macrophages
Apoptosis
Osteoclasts
Cell Death
pamidronate
In Vitro Techniques
Bone Density Conservation Agents
Etidronic Acid
Clodronic Acid
Alendronate
DNA Cleavage
Acute-Phase Reaction
Poisons
Endonucleases
Bone Diseases
Bone Resorption
Nitric Oxide Synthase
Osteoporosis

Keywords

  • Alendronate
  • Animals
  • Apoptosis
  • Bone Marrow
  • Bone Marrow Cells
  • Bone Resorption
  • Cell Division
  • Cell Nucleus
  • Cells, Cultured
  • DNA
  • DNA Fragmentation
  • Diphosphonates
  • Electrophoresis, Polyacrylamide Gel
  • Macrophages
  • Mice
  • Necrosis
  • Nitric Oxide
  • Protein Biosynthesis

Cite this

Rogers, M. J., Chilton, K. M., Coxon, F., Lawry, J., Smith, M. O., Suri, S., & Russell, R. G. G. (1996). Bisphosphonates induce apoptosis in mouse macrophage-like cells in vitro by a nitric oxide-independent mechanism. Journal of Bone and Mineral Research, 11(10), 1482-1491. https://doi.org/10.1002/jbmr.5650111015

Bisphosphonates induce apoptosis in mouse macrophage-like cells in vitro by a nitric oxide-independent mechanism. / Rogers, Michael John; Chilton, K M; Coxon, Fraser; Lawry, J; Smith, M. Olivia; Suri, S; Russell, R. Graham G. .

In: Journal of Bone and Mineral Research, Vol. 11, No. 10, 01.10.1996, p. 1482-1491.

Research output: Contribution to journalArticle

Rogers, Michael John ; Chilton, K M ; Coxon, Fraser ; Lawry, J ; Smith, M. Olivia ; Suri, S ; Russell, R. Graham G. . / Bisphosphonates induce apoptosis in mouse macrophage-like cells in vitro by a nitric oxide-independent mechanism. In: Journal of Bone and Mineral Research. 1996 ; Vol. 11, No. 10. pp. 1482-1491.
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T1 - Bisphosphonates induce apoptosis in mouse macrophage-like cells in vitro by a nitric oxide-independent mechanism

AU - Rogers, Michael John

AU - Chilton, K M

AU - Coxon, Fraser

AU - Lawry, J

AU - Smith, M. Olivia

AU - Suri, S

AU - Russell, R. Graham G.

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N2 - Bisphosphonates (BPs) are an important class of antiresorptive drugs used in the treatment of bone diseases, including osteoporosis. Although their mechanism of action has not been identified at the molecular level, there is substantial evidence that BPs can have a direct effect on osteoclasts by mechanisms that may lead to osteoclast cell death by apoptosis. BPs can also inhibit proliferation and cause cell death in macrophages in vitro. We have now shown that the toxic effect of BPs on macrophages is also due to the induction of apoptotic, rather than necrotic, cell death. Morphological and biochemical features that are definitive of apoptosis (chromatin condensation, nuclear fragmentation, and endonuclease-mediated internucleosomal cleavage of DNA) could be identified in mouse macrophage-like J774 and RAW264 cells, following treatment with 100 microM pamidronate, alendronate, and ibandronate for 24 h or more. Clodronate was much less potent, even at 2000 microM, while 2000 microM etidronate did not cause apoptosis. Apoptosis was not due to increased synthesis of nitric oxide and could not be prevented by inhibitors of nitric oxide synthases. Since macrophages, like osteoclasts, are particularly susceptible to BPs, these observations support the recent suggestion that the mechanism by which BPs inhibit bone resorption may involve osteoclast apoptosis. Furthermore, the macrophage-like cell lines used in this study may be a convenient model with which to identify the molecular mechanisms by which BPs promote apoptosis in osteoclasts. Induction of macrophage apoptosis by BPs in vivo may also account, at least in part, for the anti-inflammatory properties of BPs as well as the ability of BPs to cause an acute phase response.

AB - Bisphosphonates (BPs) are an important class of antiresorptive drugs used in the treatment of bone diseases, including osteoporosis. Although their mechanism of action has not been identified at the molecular level, there is substantial evidence that BPs can have a direct effect on osteoclasts by mechanisms that may lead to osteoclast cell death by apoptosis. BPs can also inhibit proliferation and cause cell death in macrophages in vitro. We have now shown that the toxic effect of BPs on macrophages is also due to the induction of apoptotic, rather than necrotic, cell death. Morphological and biochemical features that are definitive of apoptosis (chromatin condensation, nuclear fragmentation, and endonuclease-mediated internucleosomal cleavage of DNA) could be identified in mouse macrophage-like J774 and RAW264 cells, following treatment with 100 microM pamidronate, alendronate, and ibandronate for 24 h or more. Clodronate was much less potent, even at 2000 microM, while 2000 microM etidronate did not cause apoptosis. Apoptosis was not due to increased synthesis of nitric oxide and could not be prevented by inhibitors of nitric oxide synthases. Since macrophages, like osteoclasts, are particularly susceptible to BPs, these observations support the recent suggestion that the mechanism by which BPs inhibit bone resorption may involve osteoclast apoptosis. Furthermore, the macrophage-like cell lines used in this study may be a convenient model with which to identify the molecular mechanisms by which BPs promote apoptosis in osteoclasts. Induction of macrophage apoptosis by BPs in vivo may also account, at least in part, for the anti-inflammatory properties of BPs as well as the ability of BPs to cause an acute phase response.

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KW - Apoptosis

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KW - Bone Marrow Cells

KW - Bone Resorption

KW - Cell Division

KW - Cell Nucleus

KW - Cells, Cultured

KW - DNA

KW - DNA Fragmentation

KW - Diphosphonates

KW - Electrophoresis, Polyacrylamide Gel

KW - Macrophages

KW - Mice

KW - Necrosis

KW - Nitric Oxide

KW - Protein Biosynthesis

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DO - 10.1002/jbmr.5650111015

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VL - 11

SP - 1482

EP - 1491

JO - Journal of Bone and Mineral Research

JF - Journal of Bone and Mineral Research

SN - 0884-0431

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ER -