Abe Masashi, Takahashi Masaaki, Nagano Akira
Department of Orthopaedic Surgery, Hamamatsu University School of Medicine, 1-20-1 Handayama, Hamamatsu 431-3192, Japan.
J Biomed Mater Res A. 2005 Nov 1;75(2):494-9. doi: 10.1002/jbm.a.30452.
Hyaluronic acid (HA) is a component of the extracellular matrix of cartilage and has various effects on three-dimensional cultured chondrocytes. We measured Pyridinoline (Pyr), which is a crosslink of collagen in cultured chondrocyte-collagen composites treated with HA of different molecular weights to investigate the effects of the various molecular weights on collagen crosslink synthesis. The control group was collagen gel without cells; group N was treated without HA; and the others were treated with HA with an average molecular weight of 2.3 x10(6) Da (group H), 8.0 x10(5) Da (group M), and 2.3 x10(4) Da (group L). In the control group, the Pyr content decreased, at week 4, being one-tenth that of preculture levels. In groups H and M, it was significantly greater than that in groups L and N at week 4. Pyr/hydroxyproline, which indicates the concentration of Pyr per collagen, decreased greatly in the control group at week 3. In groups H and M, it was significantly higher than that in groups L and N at week 4 and increased to 80 and 76% of normal rabbit articular cartilage, respectively. The concentration of Pyr per collagen in cultured chondrocyte-collagen composites was similar to that of normal articular cartilage in vivo, and higher molecular weight HA may have a greater effect on the maturation of collagen in the composite.
透明质酸（HA）是软骨细胞外基质的组成部分，对三维培养的软骨细胞有多种作用。我们测量了吡啶啉（Pyr），它是不同分子量HA处理的培养软骨细胞 - 胶原蛋白复合物中胶原蛋白的交联物，以研究不同分子量对胶原蛋白交联合成的影响。对照组为无细胞的胶原凝胶；N组未用HA处理；其他组分别用平均分子量为2.3×10⁶Da的HA处理（H组）、8.0×10⁵Da的HA处理（M组）和2.3×10⁴Da的HA处理（L组）。在对照组中，第4周时Pyr含量下降，仅为预培养水平的十分之一。在H组和M组中，第4周时其含量显著高于L组和N组。表示每单位胶原蛋白中Pyr浓度的Pyr/羟脯氨酸在对照组第3周时大幅下降。在H组和M组中，第4周时其显著高于L组和N组，分别增加至正常兔关节软骨的80%和76%。培养的软骨细胞 - 胶原蛋白复合物中每单位胶原蛋白的Pyr浓度与体内正常关节软骨相似，且较高分子量的HA可能对复合物中胶原蛋白的成熟有更大影响。
