Mahdy Mohamed A A, Lei Hsiao Yin, Wakamatsu Jun-Ichi, Hosaka Yoshinao Z, Nishimura Takanori
Laboratory of Basic Veterinary Science, United Graduate School of Veterinary Science, Yamaguchi University, Yamaguchi, Yamaguchi, Japan; Department of Veterinary Anatomy, Faculty of Agriculture, Tottori University, Tottori, Tottori, Japan; Department of Anatomy and Embryology, Faculty of Veterinary Medicine, South Valley University, Qena, Egypt.
Laboratory of Muscle Biology and Meat Science, Research Faculty of Agriculture, Hokkaido University, Sapporo, Hokkaido, Japan.
Ann Anat. 2015 Nov;202:18-27. doi: 10.1016/j.aanat.2015.07.002. Epub 2015 Aug 21.
In the present study, we examined muscle regeneration following two types of chemical injuries, cardiotoxin (CTX) and glycerol, in order to compare their effect on the morphological characteristics during muscle regeneration, in addition we studied the structural changes of the intramuscular connective tissue (IMCT) during the regeneration process, by scanning electron microscopy (SEM) after digestion of the cellular elements of the muscle with sodium hydroxide. Tibialis anterior (TA) muscles of adult male mice were injected either with CTX or glycerol. Muscle degeneration was greater in the CTX-injured model than in the glycerol-injured model at day 4 post injection. Muscle regeneration started at day 7 in both the CTX and glycerol models. However, the CTX-injured model showed a higher myotube density and larger myotube diameter than the glycerol-injured model at days 10 and 14 post injection. On other hand, adipocyte infiltration was detected in the glycerol-injured model. In contrast, no adipocytes could be detected in the CTX-injured model. Furthermore, ultrastructural analysis showed a significant difference in myofiber damage and regeneration between the two models. SEM of the IMCT showed a transient increase in endomysial collagen deposition at early stages of regeneration in the CTX-injured model. In contrast, glycerol-injured model showed slight endomysial collagen deposition. Our results suggest that changes in IMCT affect the efficiency of muscle regeneration. Studying the three dimensional structure of IMCT may help clinical therapies to reduce skeletal muscle fibrosis. To our knowledge this is the first time the changes in IMCT following CTX and glycerol injury using SEM-cell maceration technique have been compared.
在本研究中,我们检测了两种化学损伤(心脏毒素(CTX)和甘油)后的肌肉再生情况,以比较它们对肌肉再生过程中形态特征的影响。此外,我们通过用氢氧化钠消化肌肉细胞成分后进行扫描电子显微镜(SEM)观察,研究了再生过程中肌内结缔组织(IMCT)的结构变化。给成年雄性小鼠的胫前肌(TA)注射CTX或甘油。注射后第4天,CTX损伤模型中的肌肉变性比甘油损伤模型更严重。在CTX和甘油模型中,肌肉再生均在第7天开始。然而,在注射后第10天和第14天,CTX损伤模型的肌管密度更高,肌管直径更大。另一方面,在甘油损伤模型中检测到脂肪细胞浸润。相比之下,在CTX损伤模型中未检测到脂肪细胞。此外,超微结构分析显示,两种模型在肌纤维损伤和再生方面存在显著差异。CTX损伤模型中IMCT的SEM显示,在再生早期,肌内膜胶原沉积短暂增加。相比之下,甘油损伤模型的肌内膜胶原沉积较少。我们的结果表明,IMCT的变化会影响肌肉再生的效率。研究IMCT的三维结构可能有助于临床治疗减少骨骼肌纤维化。据我们所知,这是首次使用SEM-细胞浸渍技术比较CTX和甘油损伤后IMCT的变化。