Knight M M, Ross J M, Sherwin A F, Lee D A, Bader D L, Poole C A
Interdisciplinary Research Centre in Biomedical Materials, Queen Mary University of London, Mile End Rd, London E1 4NS, UK.
Biochim Biophys Acta. 2001 May 3;1526(2):141-6. doi: 10.1016/s0304-4165(01)00118-0.
Within articular cartilage, the chondron microenvironment will influence chondrocyte behaviour and response to loading. Chondrons were extracted from intact cartilage using either mechanical homogenisation (MC) or enzymatic digestion (EC) and cell and matrix morphology in unstrained and compressed agarose constructs was examined. Isolated chondrocytes (IC) were used for comparison. Immunolocalisation of type VI collagen and keratan sulphate revealed differences in the structure of the pericellular microenvironment such that MC most closely resembled chondrons in situ. The unstrained cell diameters of IC and EC were larger than MC at day 1 and increased significantly over a 7 day culture period. In contrast, cell diameters for MC remained constant. Compression of constructs at day 1 resulted in cell deformation for IC and EC but not MC. The two chondron extraction methods yielded chondrons of differing matrix morphology and associated differences in cell size and cellular response to load. The results indicate that the pericellular microenvironment of MC initially possessed a greater mechanical integrity than that of EC. Although these differences may be reduced with time in culture, characterisation of mechanically isolated chondrons suggests that the stiffness of the chondrons in situ may be greater than previous estimates.
在关节软骨内，软骨细胞微环境会影响软骨细胞的行为以及对负荷的反应。使用机械匀浆法（MC）或酶消化法（EC）从完整软骨中提取软骨细胞团，并检测未受应变和受压的琼脂糖构建物中的细胞和基质形态。分离的软骨细胞（IC）用作对照。VI型胶原蛋白和硫酸角质素的免疫定位揭示了细胞周围微环境结构的差异，使得MC最接近原位软骨细胞团。第1天时，IC和EC的未受应变细胞直径大于MC，并且在7天的培养期内显著增加。相比之下，MC的细胞直径保持不变。第1天对构建物进行压缩导致IC和EC的细胞变形，但MC未变形。两种软骨细胞团提取方法产生了基质形态不同的软骨细胞团，以及细胞大小和细胞对负荷反应的相关差异。结果表明，MC的细胞周围微环境最初比EC具有更高的机械完整性。尽管这些差异可能会随着培养时间的延长而减小，但对机械分离的软骨细胞团的表征表明，原位软骨细胞团的硬度可能比先前估计的更大。
