牙本质内Ⅰ型胶原纤维形态的免疫标记初探

目的初步探讨进行免疫标记的牙本质脱矿后暴露的Ⅰ型胶原纤维。方法利用免疫组化技术和场发射扫描电镜(FEISEM)相结合,对暴露胶原纤维的形态以及结构完整性进行评价。结果通过FEISEM观察,牙本质脱矿后暴露的Ⅰ型胶原纤维,可以结合抗Ⅰ型胶原单克隆抗体以及胶体金标记的相应二抗,胶体金标记颗粒分布均匀,可与暴露的胶原纤维上的抗原表位相结合,实现免疫定位。结论免疫组化技术和FEISEM相结合,可以直观观察胶原纤维立体网络的超微结构,是一种准确的、可复制的、可行的方法 。

Preliminary research on immune labelling of type I coIIagen fibril in dentin

Objective This study was designed to evaluate the morphology of typeⅠ collagen in demineralized dentin using an immunocytochemical technique.Methods After dentin was etched with 37% phosphoric acid gel for 15 s,a double immune labeling procedure was applied,and then the morphological and structural integrity evaluation of exposed collagen fibril was carried out under a field emission in-lens scanning electron microscope (FEISEM).Results TypeⅠ collagen was labeled with secondary antibodies conjugated with 20 nm colloidal gold particles.The intense and uniform labeling signal was detected on the major fibrils indicating that the anti-typeⅠ collagen antibody could bind with favorable interaction sites on the exposed typeⅠ collagen molecular surface.Conclusions The ultrastructure of three-dimensional collagen fibril network could be identified by immunocytochemistry technology under FEISEM observation,which represented an accurate,reproducible and feasible method.