Activation of a chondrocyte volume-sensitive Cl(-) conductance prior to macroscopic cartilage lesion formation in the rabbit knee anterior cruciate ligament transection osteoarthritis model.
The anterior cruciate ligament transection (ACLT) rabbit osteoarthritis (OA) model confers permanent knee instability and induces joint degeneration. The degeneration process is complex, but includes chondrocyte apoptosis and OA-like loss of cartilage integrity. Previously, we reported that activation of a volume-sensitive Cl(-) current (ICl,vol) can mediate cell shrinkage and apoptosis in rabbit articular chondrocytes. Our objective was therefore to investigate whether ICl,vol was activated in the early stages of the rabbit ACLT OA model.



Adult Rabbits underwent unilateral ACLT and contralateral arthrotomy (sham) surgery. Rabbits were euthanized at 2 or 4 weeks. Samples were analyzed histologically and with assays of cell volume, apoptosis and electrophysiological characterization of ICl,vol.



At 2 and 4 weeks post ACLT cartilage appeared histologically normal, nevertheless cell swelling and caspase 3/7 activity were both significantly increased compared to sham controls. In cell-volume experiments, exposure of chondrocytes to hypotonic solution led to a greater increase in cell size in ACLT compared to controls. Caspase-3/7 activity, an indicator of apoptosis, was elevated in both ACLT 2wk and 4wk. Whole-cell currents were recorded with patch clamp of chondrocytes in iso-osmotic and hypo-osmotic external solutions under conditions where Na(+), K(+) and Ca(2+) currents were minimized. ACLT treatment resulted in a large increase in hypotonic-activated chloride conductance.



Changes in chondrocyte ion channels take place prior to the onset of apparent cartilage loss in the ACLT rabbit model of OA. Further studies are needed to investigate if pharmacological inhibition of ICl,vol decreases progression of OA in animal models.
Department of Musculoskeletal Biology, Institute of Aging and Chronic Disease, University of Liverpool, UK; Department of Orthopedic Surgery, Shiga University of Medical Science, Japan.
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Kumagai
Department of Physiology, Shiga University of Medical Science, Japan.
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Toyoda
Department of Musculoskeletal Biology, Institute of Aging and Chronic Disease, University of Liverpool, UK.
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Staunton
Department of Orthopedic Surgery, Shiga University of Medical Science, Japan.
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Maeda
Department of Orthopedic Surgery, Shiga University of Medical Science, Japan.
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Okumura
Department of Physiology, Shiga University of Medical Science, Japan.
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Matsuura
Department of Orthopedic Surgery, Shiga University of Medical Science, Japan.
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Matsusue
Department of Orthopedic Surgery, Shiga University of Medical Science, Japan.
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Imai
Department of Musculoskeletal Biology, Institute of Aging and Chronic Disease, University of Liverpool, UK. Electronic address: rbj@liverpool.ac.uk.
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Barrett-Jolley
D000818:Animals; D016118:Anterior Cruciate Ligament; D000070598:Anterior Cruciate Ligament Injuries; D002358:Cartilage, Articular; D019902:Chondrocytes; D004195:Disease Models, Animal; D010003:Osteoarthritis; D020370:Osteoarthritis, Knee; D011817:Rabbits
ACLT; Anion channel; Apoptosis; Cartilage; Caspase; Chloride channel; Ion channels; Osteoarthritis