Mello M L S, de Campos Vidal B
Department of Cell Biology, Institute of Biology, UNICAMP, 13084-971 Campinas, SP, Brazil.
Cell Mol Biol (Noisy-le-grand). 2003 Jun;49(4):579-85.
Changes in the macromolecular orientation and metachromasy of glycosaminoglycans (GAG) in newly synthesized and assembled collagen fibers in rat Achilles tendon after tendon excision were investigated in toluidine blue (TB)-stained preparations, based in the selective absorption of polarized light (= linear dichroism, LD) and of absorption of unpolarized light in situ. Extrinsic LD was observed microspectrophotometrically from the early phases of tendon repair onwards, although the absorption peaks in both parallel and perpendicular directions with respect to the plane of polarized light and the long axis of the collagen fibers occurred at the same wavelength, and thus differed from the pattern situation in normal adult controls. Compared to normal adult tendons, the pattern of LD in newly synthesized and assembled fibers was still not fully attained 110 days after surgical tendon removal. This incomplete recovery possibly reflected the influence of aging during the repair process. There was no correlation between LD and metachromasy. The highest absorption values for metachromatic staining occurred on the 7th day after tendon removal, at a time when LD was not intense. Treatment with hyaluronidase showed that the LD in the early stages of tendon repair was mostly due to hyaluronate whereas the LD in the later stages was due to chondroitin sulfates. The changes in LD during Achilles tendon repair were attributed to gradual modifications in the composition and macromolecular orientation of GAGs relative to the long axis of the collagen fibers.
基于偏振光的选择性吸收(= 线性二色性,LD)和原位非偏振光的吸收,在甲苯胺蓝(TB)染色制剂中研究了大鼠跟腱切除后新合成和组装的胶原纤维中糖胺聚糖(GAG)的大分子取向和异染性变化。从肌腱修复的早期阶段开始,用显微分光光度计观察到外在LD,尽管相对于偏振光平面和胶原纤维长轴的平行和垂直方向上的吸收峰都出现在相同波长处,因此与正常成年对照中的模式情况不同。与正常成年肌腱相比,手术切除肌腱110天后,新合成和组装纤维中的LD模式仍未完全达到。这种不完全恢复可能反映了修复过程中衰老的影响。LD与异染性之间没有相关性。异染染色的最高吸收值出现在肌腱切除后的第7天,此时LD并不强烈。用透明质酸酶处理表明,肌腱修复早期的LD主要归因于透明质酸盐,而后期的LD则归因于硫酸软骨素。跟腱修复过程中LD的变化归因于GAGs相对于胶原纤维长轴的组成和大分子取向的逐渐改变。