Annexin 5 overexpression increased articular chondrocyte apoptosis induced by basic calcium phosphate crystals

H. K. Ea, V. Monceau, Emmanuel Camors, M. Cohen-Solal, D. Charlemagne, Frédéric Lioté

Research output: Contribution to journalArticle

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Abstract

Objectives: Basic calcium phosphate (BCP) crystals (octacalcium phosphate (OCP), carbapatite (CA) and hydroxyapatite (HA)) are associated with severe forms of osteoarthritis. In advanced osteoarthritis, cartilage shows chondrocyte apoptosis, overexpression of annexin 5 (A5) and BCP crystal deposition within matrix vesicles. We assessed in vitro whether BCP crystals and overexpression of A5 increased chondrocyte apoptosis. Methods: Apoptosis was induced by BCP crystals, tumour necrosis factor (TNF)-α (20 ng/ml) and Fas ligand (20 ng/ml) in normal articular chondrocytes (control) and in A5 overexpressed chondrocytes, performed by adenovirus infection. Apoptosis was assessed by caspase 3 (Cas3) activity, and DNA fragmentation. Results: All BCP crystals, TNF-α and Fas ligand induced chondrocyte apoptosis as demonstrated by decreased cell viability and increased Cas3 activity and DNA fragmentation. TUNEL (terminal deoxyribonucleotide transferase-mediated dUTP nick end-labelling)-positive staining chondrocytes were increased by OCP (12.4 (5.2)%), CA (9.6 (2.6)%) and HA (9.2 (3.0)%) crystals and TNF-α (9.6 (2.4)%) stimulation compared with control (3.1 (1.9)%). BCP crystals increased Cas3 activity in a dose-dependent fashion. BCP-crystal-induced chondrocyte apoptosis was independent from TNF-α and interleukin-1β pathways but required cell-crystal contact and intralysosomal crystal dissolution. Indeed, preincubation with ammonium chloride, a lysosomal inhibitor of BCP crystal dissolution, significantly decreased BCP-crystal-induced Cas3 activity. Finally, overexpression of A5 enhanced BCP crystal- and TNF-α-induced chondrocyte apoptosis. Conclusions: Overexpression of A5 and the presence of BCP crystals observed in advanced osteoarthritis contributed to chondrocyte apoptosis. Our results suggest a new pathophysiological mechanism for calcium-containing crystal arthropathies.

Original languageEnglish (US)
Pages (from-to)1617-1625
Number of pages9
JournalAnnals of the Rheumatic Diseases
Volume67
Issue number11
DOIs
StatePublished - Nov 1 2008
Externally publishedYes

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Annexins
Chondrocytes
Joints
Apoptosis
Crystals
Tumor Necrosis Factor-alpha
Caspase 3
Osteoarthritis
Fas Ligand Protein
DNA Fragmentation
Durapatite
calcium phosphate
Deoxyribonucleotides
Adenoviridae Infections
Ammonium Chloride
Dissolution
Transferases
Interleukin-1
Cartilage
Cell Survival

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Rheumatology
  • Immunology
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Annexin 5 overexpression increased articular chondrocyte apoptosis induced by basic calcium phosphate crystals. / Ea, H. K.; Monceau, V.; Camors, Emmanuel; Cohen-Solal, M.; Charlemagne, D.; Lioté, Frédéric.

In: Annals of the Rheumatic Diseases, Vol. 67, No. 11, 01.11.2008, p. 1617-1625.

Research output: Contribution to journalArticle

Ea, H. K. ; Monceau, V. ; Camors, Emmanuel ; Cohen-Solal, M. ; Charlemagne, D. ; Lioté, Frédéric. / Annexin 5 overexpression increased articular chondrocyte apoptosis induced by basic calcium phosphate crystals. In: Annals of the Rheumatic Diseases. 2008 ; Vol. 67, No. 11. pp. 1617-1625.
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abstract = "Objectives: Basic calcium phosphate (BCP) crystals (octacalcium phosphate (OCP), carbapatite (CA) and hydroxyapatite (HA)) are associated with severe forms of osteoarthritis. In advanced osteoarthritis, cartilage shows chondrocyte apoptosis, overexpression of annexin 5 (A5) and BCP crystal deposition within matrix vesicles. We assessed in vitro whether BCP crystals and overexpression of A5 increased chondrocyte apoptosis. Methods: Apoptosis was induced by BCP crystals, tumour necrosis factor (TNF)-α (20 ng/ml) and Fas ligand (20 ng/ml) in normal articular chondrocytes (control) and in A5 overexpressed chondrocytes, performed by adenovirus infection. Apoptosis was assessed by caspase 3 (Cas3) activity, and DNA fragmentation. Results: All BCP crystals, TNF-α and Fas ligand induced chondrocyte apoptosis as demonstrated by decreased cell viability and increased Cas3 activity and DNA fragmentation. TUNEL (terminal deoxyribonucleotide transferase-mediated dUTP nick end-labelling)-positive staining chondrocytes were increased by OCP (12.4 (5.2){\%}), CA (9.6 (2.6){\%}) and HA (9.2 (3.0){\%}) crystals and TNF-α (9.6 (2.4){\%}) stimulation compared with control (3.1 (1.9){\%}). BCP crystals increased Cas3 activity in a dose-dependent fashion. BCP-crystal-induced chondrocyte apoptosis was independent from TNF-α and interleukin-1β pathways but required cell-crystal contact and intralysosomal crystal dissolution. Indeed, preincubation with ammonium chloride, a lysosomal inhibitor of BCP crystal dissolution, significantly decreased BCP-crystal-induced Cas3 activity. Finally, overexpression of A5 enhanced BCP crystal- and TNF-α-induced chondrocyte apoptosis. Conclusions: Overexpression of A5 and the presence of BCP crystals observed in advanced osteoarthritis contributed to chondrocyte apoptosis. Our results suggest a new pathophysiological mechanism for calcium-containing crystal arthropathies.",
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T1 - Annexin 5 overexpression increased articular chondrocyte apoptosis induced by basic calcium phosphate crystals

AU - Ea, H. K.

AU - Monceau, V.

AU - Camors, Emmanuel

AU - Cohen-Solal, M.

AU - Charlemagne, D.

AU - Lioté, Frédéric

PY - 2008/11/1

Y1 - 2008/11/1

N2 - Objectives: Basic calcium phosphate (BCP) crystals (octacalcium phosphate (OCP), carbapatite (CA) and hydroxyapatite (HA)) are associated with severe forms of osteoarthritis. In advanced osteoarthritis, cartilage shows chondrocyte apoptosis, overexpression of annexin 5 (A5) and BCP crystal deposition within matrix vesicles. We assessed in vitro whether BCP crystals and overexpression of A5 increased chondrocyte apoptosis. Methods: Apoptosis was induced by BCP crystals, tumour necrosis factor (TNF)-α (20 ng/ml) and Fas ligand (20 ng/ml) in normal articular chondrocytes (control) and in A5 overexpressed chondrocytes, performed by adenovirus infection. Apoptosis was assessed by caspase 3 (Cas3) activity, and DNA fragmentation. Results: All BCP crystals, TNF-α and Fas ligand induced chondrocyte apoptosis as demonstrated by decreased cell viability and increased Cas3 activity and DNA fragmentation. TUNEL (terminal deoxyribonucleotide transferase-mediated dUTP nick end-labelling)-positive staining chondrocytes were increased by OCP (12.4 (5.2)%), CA (9.6 (2.6)%) and HA (9.2 (3.0)%) crystals and TNF-α (9.6 (2.4)%) stimulation compared with control (3.1 (1.9)%). BCP crystals increased Cas3 activity in a dose-dependent fashion. BCP-crystal-induced chondrocyte apoptosis was independent from TNF-α and interleukin-1β pathways but required cell-crystal contact and intralysosomal crystal dissolution. Indeed, preincubation with ammonium chloride, a lysosomal inhibitor of BCP crystal dissolution, significantly decreased BCP-crystal-induced Cas3 activity. Finally, overexpression of A5 enhanced BCP crystal- and TNF-α-induced chondrocyte apoptosis. Conclusions: Overexpression of A5 and the presence of BCP crystals observed in advanced osteoarthritis contributed to chondrocyte apoptosis. Our results suggest a new pathophysiological mechanism for calcium-containing crystal arthropathies.

AB - Objectives: Basic calcium phosphate (BCP) crystals (octacalcium phosphate (OCP), carbapatite (CA) and hydroxyapatite (HA)) are associated with severe forms of osteoarthritis. In advanced osteoarthritis, cartilage shows chondrocyte apoptosis, overexpression of annexin 5 (A5) and BCP crystal deposition within matrix vesicles. We assessed in vitro whether BCP crystals and overexpression of A5 increased chondrocyte apoptosis. Methods: Apoptosis was induced by BCP crystals, tumour necrosis factor (TNF)-α (20 ng/ml) and Fas ligand (20 ng/ml) in normal articular chondrocytes (control) and in A5 overexpressed chondrocytes, performed by adenovirus infection. Apoptosis was assessed by caspase 3 (Cas3) activity, and DNA fragmentation. Results: All BCP crystals, TNF-α and Fas ligand induced chondrocyte apoptosis as demonstrated by decreased cell viability and increased Cas3 activity and DNA fragmentation. TUNEL (terminal deoxyribonucleotide transferase-mediated dUTP nick end-labelling)-positive staining chondrocytes were increased by OCP (12.4 (5.2)%), CA (9.6 (2.6)%) and HA (9.2 (3.0)%) crystals and TNF-α (9.6 (2.4)%) stimulation compared with control (3.1 (1.9)%). BCP crystals increased Cas3 activity in a dose-dependent fashion. BCP-crystal-induced chondrocyte apoptosis was independent from TNF-α and interleukin-1β pathways but required cell-crystal contact and intralysosomal crystal dissolution. Indeed, preincubation with ammonium chloride, a lysosomal inhibitor of BCP crystal dissolution, significantly decreased BCP-crystal-induced Cas3 activity. Finally, overexpression of A5 enhanced BCP crystal- and TNF-α-induced chondrocyte apoptosis. Conclusions: Overexpression of A5 and the presence of BCP crystals observed in advanced osteoarthritis contributed to chondrocyte apoptosis. Our results suggest a new pathophysiological mechanism for calcium-containing crystal arthropathies.

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