Duplan-Perrat F, Damour O, Montrocher C, Peyrol S, Grenier G, Jacob M P, Braye F
Laboratoire des Substituts Cutanés, Service de biochimie C, Hôpital E. Herriot, Lyon, France.
J Invest Dermatol. 2000 Feb;114(2):365-70. doi: 10.1046/j.1523-1747.2000.00885.x.
Elastic fibers form a complex network that contributes to the elasticity of connective tissues. Alterations in the elastic fiber network are involved in several disease affecting organs in which compliance of the connective tissue is essential: skin, main vasculature, lung, joints, muscle, and ligament. The aim of our work was to study the deposition, maturation, and organization of elastic fiber components in a dermal equivalent model consisting of collagen-GAG-chitosan seeded with fibroblasts. The influence of keratinocytes was studied in parallel, thus constituting a skin equivalent model. These models were examined by transmission electron microscopy (TEM) and by immunohistochemistry to determine the staining patterns of fibrillin-1 and elastin proteins representative of the microfibrillar framework and of the elastic fibers, respectively. After 2 mo of fibroblast culture in the dermal equivalent, elastin was undetectable, whereas fibrillin-1 staining was weak and microfibrils were infrequently observed by TEM. In the skin equivalent, fibrillin-1 and elastin were detected by immunostaining 15 d after epidermization and TEM revealed the typical structure and organization of the elastic network in the dermis, with elastin deposition on the microfibrillar scaffold. This in vitro skin equivalent model is to our knowledge the first in which elastic fibers have been detected, thus demonstrating the influence of keratinocytes on the maturation and organization of the elastic network.
弹性纤维形成一个复杂的网络,有助于结缔组织的弹性。弹性纤维网络的改变与几种影响器官的疾病有关,在这些疾病中,结缔组织的顺应性至关重要,如皮肤、主要脉管系统、肺、关节、肌肉和韧带。我们研究的目的是在由接种成纤维细胞的胶原蛋白-糖胺聚糖-壳聚糖组成的真皮替代模型中,研究弹性纤维成分的沉积、成熟和组织情况。同时研究了角质形成细胞的影响,从而构建了一个皮肤替代模型。通过透射电子显微镜(TEM)和免疫组织化学对这些模型进行检查,以分别确定代表微原纤维框架和弹性纤维的原纤蛋白-1和弹性蛋白的染色模式。在真皮替代物中培养成纤维细胞2个月后,未检测到弹性蛋白,而原纤蛋白-1染色较弱,通过TEM很少观察到微原纤维。在皮肤替代物中,表皮化15天后通过免疫染色检测到原纤蛋白-1和弹性蛋白,TEM显示真皮中弹性网络的典型结构和组织,弹性蛋白沉积在微原纤维支架上。据我们所知,这个体外皮肤替代模型是第一个检测到弹性纤维的模型,从而证明了角质形成细胞对弹性网络成熟和组织的影响。