Calcium Homeostasis in Articular Chondrocytes of Two Different Animal Species

R. White, J. Gibson
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Abstract

Introduction: Intracellular calcium concentration ([Ca2+]i) is a critical parameter in cellular homeostasis, including articular chondrocytes. Perturbed [Ca2+]i of chondrocytes may be associated with joint disease. The objective of the study was to compare large animal models for investigating Ca2+ homeostasis in chondrocytes. Materials and Methods: The gross anatomy of the metacarpophalangeal joint (MCP) of cattle and sheep was compared, along with the effect of various manoeuvres used to study the mechanisms of Ca2+ homeostasis in chondrocytes from load-bearing areas. The gross anatomy was observed before and after dissection, and internal architecture was examined after sectioning. Cartilage thickness was measured with a digital micrometer. Chondrocyte yield was determined after isolation. Chondrocytes were incubated with Fura-2 and Ca2+i followed in different extracellular conditions. A hypotonic shock (HTS) was used to mimic removal of a load. Results: The results showed that ovids and bovids were skeletally immature and aspects of Ca2+ homeostasis were similar. Ovine chondrocytes had higher resting fluorescence, consistent with elevated resting Ca2+ levels. Results from ion substitution experiments were consistent with a role for Na+/Ca2+ exchange, and swelling-induced Ca2+ enters into the cytoplasm via the plasma membrane and intracellular stores. Conclusions: Ca2+ homeostasis in chondrocytes from both species behaved in a similar manner to HTS and ion substitutions. Differences in resting [Ca2+]i could be associated with species, stage of maturation, or Fura-2 itself and require further investigation. These findings contribute to our understanding of the physiology of articular cartilage in different species, and their potential use as models for studying joint disease in humans.
两种不同动物关节软骨细胞钙稳态的研究
细胞内钙浓度([Ca2+]i)是细胞稳态的关键参数,包括关节软骨细胞。软骨细胞的[Ca2+]i紊乱可能与关节疾病有关。该研究的目的是比较大型动物模型来研究软骨细胞中的Ca2+稳态。材料和方法:比较了牛和羊掌指关节(MCP)的大体解剖结构,以及用于研究负重区软骨细胞Ca2+稳态机制的各种操作的影响。解剖前后观察大体解剖,切片后检查内部结构。用数字千分尺测量软骨厚度。分离后测定软骨细胞产量。软骨细胞与Fura-2和Ca2+i在不同的细胞外条件下孵育。低渗休克(HTS)被用来模拟负荷的移除。结果:结果表明,卵细胞和牛细胞骨骼发育不成熟,Ca2+稳态方面相似。羊软骨细胞有较高的静息荧光,与升高的静息Ca2+水平一致。离子替代实验的结果与Na+/Ca2+交换的作用一致,肿胀诱导的Ca2+通过质膜和细胞内储存进入细胞质。结论:两种动物软骨细胞中的Ca2+稳态与HTS和离子取代的行为方式相似。静息[Ca2+]i的差异可能与物种、成熟阶段或Fura-2本身有关,需要进一步研究。这些发现有助于我们了解不同物种关节软骨的生理学,以及它们作为研究人类关节疾病模型的潜在用途。
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