Shikh Alsook M K, Gabriel A, Salouci M, Piret J, Alzamel N, Moula N, Denoix J-M, Antoine N, Baise E
Anatomy Unit, FARAH Research Centre and Faculty of Veterinary Medicine, University of Liège, Liège, Belgium.
Anatomy Unit, FARAH Research Centre and Faculty of Veterinary Medicine, University of Liège, Liège, Belgium.
Vet J. 2015 Apr;204(1):117-22. doi: 10.1016/j.tvjl.2015.02.011. Epub 2015 Feb 12.
Suspensory ligament (SL) injuries are an important cause of lameness in horses. The mechanical properties of connective tissue in normal and pathological ligaments are mainly related to fibril morphology, as well as collagen content and types. The purpose of this study was to evaluate, using biochemical and ultrastructural approaches, the alterations in collagen fibrils after injury. Eight Warmblood horses with visible signs of injury in only one forelimb SL were selected and specimens were examined by transmission electron microscope (TEM). Collagen types I, III and V were purified by differential salt precipitation after collagen extraction with acetic acid containing pepsin. TEM revealed abnormal organization as well as alterations in the diameter and shape of fibrils after SL injury. The bands corresponding to types I, III and V collagen were assessed by densitometry after sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Densitometric analysis indicated that the proportions of type III and type V collagen were higher (P < 0.001) in damaged tissues compared with normal tissues with a mean increase of 20.9% and 17.3%, respectively. Concurrently, a decrease (P < 0.001) in type I collagen within damaged tissues was recorded with a mean decrease of 15.2%. These alterations could be the hallmark of a decrease in the tissue quality and mechanical properties of the ligament. The findings provide new insight for subsequent research on tissue regeneration that may lead to the development of future treatment strategies for SL injury.
悬韧带（SL）损伤是马匹跛行的一个重要原因。正常和病理状态下韧带中结缔组织的力学特性主要与纤维形态以及胶原蛋白的含量和类型有关。本研究的目的是采用生化和超微结构方法评估损伤后胶原纤维的变化。选择8匹仅一侧前肢悬韧带出现明显损伤迹象的温血马，并通过透射电子显微镜（TEM）检查标本。用含胃蛋白酶的乙酸提取胶原蛋白后，通过分级盐沉淀法纯化I型、III型和V型胶原蛋白。透射电子显微镜显示，悬韧带损伤后纤维的组织结构异常，直径和形状也发生了改变。在十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）后，通过光密度测定法评估对应I型、III型和V型胶原蛋白的条带。光密度分析表明，与正常组织相比，受损组织中III型和V型胶原蛋白的比例更高（P < 0.001），平均分别增加了20.9%和17.3%。同时，记录到受损组织中I型胶原蛋白减少（P < 0.001），平均减少了15.2%。这些变化可能是韧带组织质量和力学性能下降的标志。这些发现为后续关于组织再生的研究提供了新的见解，可能会促成未来悬韧带损伤治疗策略的发展。
