Nonsteroidal anti-inflammatory drugs arrest cell cycle in G0/G1 phase and induce cell death in osteoblast-enriched cultures

Mei Ling Ho, Je Ken Chang, Hsiu Ting Tsai, Ming Hsuang Cho, Gwo Jaw Wang

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Nonsteroidal anti-inflammatory drugs have been widely prescribed for orthopaedic patients to relieve pain and chronic inflammation. However, it has been demonstrated that NSAIDs suppress bone repair and remodeling in vivo. We have reported that ketorolac inhibits bone repair in vivo and its critical effective timing is at the early stage of endochondral ossification. Our previous results showed that ketorolac and indomethacin inhibit osteoblast proliferation in vitro, suggesting that this effect may be one of the mechanisms contributing to the suppressive effect of NSAIDs on bone remodeling. Cell proliferation and death of osteoblasts should be well regulated through some relative apoptotic and mitotic factors during normal bone remodeling process. Accordingly, we proposed that the induction of osteoblastic cell death of NSAIDs might be one of the mechanisms involving their suppressive effect on bone remodeling in vivo. In this study, we investigated whether NSAIDs arrest osteoblastic cell cycle and/or induce cell death. Whether the mechanism was mediated through prostaglandin (PG) pathway. We tested the effects of ketorolac, indomethacin, diclofenac, piroxicam on cell cycle kinetics, cytotoxicity, and cell death pattern in osteoblast-enriched cultures derived from fetal rat calvaria. Our results showed that ketorolac and indomethacin arrested cell cycle at G0/G1 phase. All the 4 NSAIDs had cytotoxic effects and these effects were concentration dependent. The sequence of the cytotoxic effects of these four NSAIDs at 10-4 M were indomethacin > diclofenac > ketorolac > piroxicam. Both PGE1 and PGE2 (10-10-10-8 M) also significantly elevated the LDH leakage of osteoblasts, while PGF had no significant effect. These results revealed that the cytotoxic effects of NSAIDs on osteoblasts might not be through inhibiting prostaglandin synthesis. They may be through PG-independent pathways. The results from flow cytometry followed by AnnexinV-FITC and propidium iodide double staining showed that 24 hours treatment of all the 4 NSAIDs (10-6 and 10-4 M) significantly induced both apoptosis (p <0.01) and necrosis (p <0.01, or p <0.05) in osteoblast cultures. These effects of NSAIDs on cell cycle arrest and cell death induction in osteoblasts may be one of the important mechanisms contributing to their suppressive effect on bone repair and bone remodeling in vivo.

Original languageEnglish
Pages (from-to)279-289
Number of pages11
JournalJournal of Musculoskeletal Research
Volume5
Issue number4
DOIs
Publication statusPublished - 2001
Externally publishedYes

Fingerprint

Cell Cycle Resting Phase
G1 Phase
Non-Steroidal Anti-Inflammatory Agents
Cell Cycle Checkpoints
Osteoblasts
Cell Death
Anti-Inflammatory Agents
Ketorolac
Bone Remodeling
Pharmaceutical Preparations
Indomethacin
Prostaglandins
Piroxicam
Diclofenac
Cell Cycle
Bone and Bones
Dinoprost
Propidium
Fluorescein-5-isothiocyanate
Alprostadil

Keywords

  • Apoptosis
  • Necrosis
  • NSAIDs
  • Osteoblasts

ASJC Scopus subject areas

  • Surgery

Cite this

Nonsteroidal anti-inflammatory drugs arrest cell cycle in G0/G1 phase and induce cell death in osteoblast-enriched cultures. / Ho, Mei Ling; Chang, Je Ken; Tsai, Hsiu Ting; Cho, Ming Hsuang; Wang, Gwo Jaw.

In: Journal of Musculoskeletal Research, Vol. 5, No. 4, 2001, p. 279-289.

Research output: Contribution to journalArticle

@article{92d09042621a4ee19bcfb9c8821c320d,
title = "Nonsteroidal anti-inflammatory drugs arrest cell cycle in G0/G1 phase and induce cell death in osteoblast-enriched cultures",
abstract = "Nonsteroidal anti-inflammatory drugs have been widely prescribed for orthopaedic patients to relieve pain and chronic inflammation. However, it has been demonstrated that NSAIDs suppress bone repair and remodeling in vivo. We have reported that ketorolac inhibits bone repair in vivo and its critical effective timing is at the early stage of endochondral ossification. Our previous results showed that ketorolac and indomethacin inhibit osteoblast proliferation in vitro, suggesting that this effect may be one of the mechanisms contributing to the suppressive effect of NSAIDs on bone remodeling. Cell proliferation and death of osteoblasts should be well regulated through some relative apoptotic and mitotic factors during normal bone remodeling process. Accordingly, we proposed that the induction of osteoblastic cell death of NSAIDs might be one of the mechanisms involving their suppressive effect on bone remodeling in vivo. In this study, we investigated whether NSAIDs arrest osteoblastic cell cycle and/or induce cell death. Whether the mechanism was mediated through prostaglandin (PG) pathway. We tested the effects of ketorolac, indomethacin, diclofenac, piroxicam on cell cycle kinetics, cytotoxicity, and cell death pattern in osteoblast-enriched cultures derived from fetal rat calvaria. Our results showed that ketorolac and indomethacin arrested cell cycle at G0/G1 phase. All the 4 NSAIDs had cytotoxic effects and these effects were concentration dependent. The sequence of the cytotoxic effects of these four NSAIDs at 10-4 M were indomethacin > diclofenac > ketorolac > piroxicam. Both PGE1 and PGE2 (10-10-10-8 M) also significantly elevated the LDH leakage of osteoblasts, while PGF2α had no significant effect. These results revealed that the cytotoxic effects of NSAIDs on osteoblasts might not be through inhibiting prostaglandin synthesis. They may be through PG-independent pathways. The results from flow cytometry followed by AnnexinV-FITC and propidium iodide double staining showed that 24 hours treatment of all the 4 NSAIDs (10-6 and 10-4 M) significantly induced both apoptosis (p <0.01) and necrosis (p <0.01, or p <0.05) in osteoblast cultures. These effects of NSAIDs on cell cycle arrest and cell death induction in osteoblasts may be one of the important mechanisms contributing to their suppressive effect on bone repair and bone remodeling in vivo.",
keywords = "Apoptosis, Necrosis, NSAIDs, Osteoblasts",
author = "Ho, {Mei Ling} and Chang, {Je Ken} and Tsai, {Hsiu Ting} and Cho, {Ming Hsuang} and Wang, {Gwo Jaw}",
year = "2001",
doi = "10.1142/S0218957701000623",
language = "English",
volume = "5",
pages = "279--289",
journal = "Journal of Musculoskeletal Research",
issn = "0218-9577",
publisher = "World Scientific Publishing Co. Pte Ltd",
number = "4",

}

TY - JOUR

T1 - Nonsteroidal anti-inflammatory drugs arrest cell cycle in G0/G1 phase and induce cell death in osteoblast-enriched cultures

AU - Ho, Mei Ling

AU - Chang, Je Ken

AU - Tsai, Hsiu Ting

AU - Cho, Ming Hsuang

AU - Wang, Gwo Jaw

PY - 2001

Y1 - 2001

N2 - Nonsteroidal anti-inflammatory drugs have been widely prescribed for orthopaedic patients to relieve pain and chronic inflammation. However, it has been demonstrated that NSAIDs suppress bone repair and remodeling in vivo. We have reported that ketorolac inhibits bone repair in vivo and its critical effective timing is at the early stage of endochondral ossification. Our previous results showed that ketorolac and indomethacin inhibit osteoblast proliferation in vitro, suggesting that this effect may be one of the mechanisms contributing to the suppressive effect of NSAIDs on bone remodeling. Cell proliferation and death of osteoblasts should be well regulated through some relative apoptotic and mitotic factors during normal bone remodeling process. Accordingly, we proposed that the induction of osteoblastic cell death of NSAIDs might be one of the mechanisms involving their suppressive effect on bone remodeling in vivo. In this study, we investigated whether NSAIDs arrest osteoblastic cell cycle and/or induce cell death. Whether the mechanism was mediated through prostaglandin (PG) pathway. We tested the effects of ketorolac, indomethacin, diclofenac, piroxicam on cell cycle kinetics, cytotoxicity, and cell death pattern in osteoblast-enriched cultures derived from fetal rat calvaria. Our results showed that ketorolac and indomethacin arrested cell cycle at G0/G1 phase. All the 4 NSAIDs had cytotoxic effects and these effects were concentration dependent. The sequence of the cytotoxic effects of these four NSAIDs at 10-4 M were indomethacin > diclofenac > ketorolac > piroxicam. Both PGE1 and PGE2 (10-10-10-8 M) also significantly elevated the LDH leakage of osteoblasts, while PGF2α had no significant effect. These results revealed that the cytotoxic effects of NSAIDs on osteoblasts might not be through inhibiting prostaglandin synthesis. They may be through PG-independent pathways. The results from flow cytometry followed by AnnexinV-FITC and propidium iodide double staining showed that 24 hours treatment of all the 4 NSAIDs (10-6 and 10-4 M) significantly induced both apoptosis (p <0.01) and necrosis (p <0.01, or p <0.05) in osteoblast cultures. These effects of NSAIDs on cell cycle arrest and cell death induction in osteoblasts may be one of the important mechanisms contributing to their suppressive effect on bone repair and bone remodeling in vivo.

AB - Nonsteroidal anti-inflammatory drugs have been widely prescribed for orthopaedic patients to relieve pain and chronic inflammation. However, it has been demonstrated that NSAIDs suppress bone repair and remodeling in vivo. We have reported that ketorolac inhibits bone repair in vivo and its critical effective timing is at the early stage of endochondral ossification. Our previous results showed that ketorolac and indomethacin inhibit osteoblast proliferation in vitro, suggesting that this effect may be one of the mechanisms contributing to the suppressive effect of NSAIDs on bone remodeling. Cell proliferation and death of osteoblasts should be well regulated through some relative apoptotic and mitotic factors during normal bone remodeling process. Accordingly, we proposed that the induction of osteoblastic cell death of NSAIDs might be one of the mechanisms involving their suppressive effect on bone remodeling in vivo. In this study, we investigated whether NSAIDs arrest osteoblastic cell cycle and/or induce cell death. Whether the mechanism was mediated through prostaglandin (PG) pathway. We tested the effects of ketorolac, indomethacin, diclofenac, piroxicam on cell cycle kinetics, cytotoxicity, and cell death pattern in osteoblast-enriched cultures derived from fetal rat calvaria. Our results showed that ketorolac and indomethacin arrested cell cycle at G0/G1 phase. All the 4 NSAIDs had cytotoxic effects and these effects were concentration dependent. The sequence of the cytotoxic effects of these four NSAIDs at 10-4 M were indomethacin > diclofenac > ketorolac > piroxicam. Both PGE1 and PGE2 (10-10-10-8 M) also significantly elevated the LDH leakage of osteoblasts, while PGF2α had no significant effect. These results revealed that the cytotoxic effects of NSAIDs on osteoblasts might not be through inhibiting prostaglandin synthesis. They may be through PG-independent pathways. The results from flow cytometry followed by AnnexinV-FITC and propidium iodide double staining showed that 24 hours treatment of all the 4 NSAIDs (10-6 and 10-4 M) significantly induced both apoptosis (p <0.01) and necrosis (p <0.01, or p <0.05) in osteoblast cultures. These effects of NSAIDs on cell cycle arrest and cell death induction in osteoblasts may be one of the important mechanisms contributing to their suppressive effect on bone repair and bone remodeling in vivo.

KW - Apoptosis

KW - Necrosis

KW - NSAIDs

KW - Osteoblasts

UR - http://www.scopus.com/inward/record.url?scp=0035727175&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035727175&partnerID=8YFLogxK

U2 - 10.1142/S0218957701000623

DO - 10.1142/S0218957701000623

M3 - Article

VL - 5

SP - 279

EP - 289

JO - Journal of Musculoskeletal Research

JF - Journal of Musculoskeletal Research

SN - 0218-9577

IS - 4

ER -