Morzel Martine, Siying Tai, Brignot Hélène, Lherminier Jeannine
CNRS, UMR6265 Centre des Sciences du Goût et de l'Alimentation, F-21000, Dijon, France; INRA, UMR1324 Centre des Sciences du Goût et de l'Alimentation, F-21000, Dijon, France; Université de Bourgogne, UMR Centre des Sciences du Goût et de l'Alimentation, F-21000, Dijon, France.
Microsc Res Tech. 2014 Jun;77(6):453-7. doi: 10.1002/jemt.22366. Epub 2014 Apr 4.
The mucosal pellicle is defined as the protein film adsorbed onto oral mucosa. This study aimed at characterizing the ultrastructure of human epithelial buccal cells and localizing salivary mucins MUC5B, a major constituent of the mucosal pellicle. Cells were sampled from the buccal surface and prepared for Transmission Electron Microscopy using high-pressure freezing/cryosubstitution followed by immunogold labelling of MUC5B. Morphologically, cells were visualized as typical cells of the superficial layer of a squamous nonkeratinized epithelium with a partly degraded plasma membrane. The outer surface of the plasma membrane was lined with a biological material of medium electron density. MUC5B were detected in the extracellular space, and particularly in the vicinity of the plasma membrane, sometimes onto fibrils protruding from the membrane. This area was, therefore, considered as constituting the mucosal pellicle, which appeared as a mixed film of both salivary and epithelial components. The distribution of gold particles suggested that the surface of the pellicle was not uniform, and that the film thickness could reach up to 100 nm. This work showed the feasibility of visualizing and characterizing the mucosal pellicle directly on human epithelial buccal cells sampled in a noninvasive manner.
黏膜薄膜被定义为吸附在口腔黏膜上的蛋白质膜。本研究旨在表征人类颊上皮细胞的超微结构,并定位唾液黏蛋白MUC5B,它是黏膜薄膜的主要成分。从颊面采集细胞,采用高压冷冻/冷冻置换法制备用于透射电子显微镜观察的样本,随后对MUC5B进行免疫金标记。在形态学上,细胞呈现为鳞状非角化上皮表层的典型细胞,质膜部分降解。质膜的外表面衬有中等电子密度的生物材料。在细胞外空间检测到了MUC5B,特别是在质膜附近,有时在从膜突出的纤维上也能检测到。因此,该区域被认为构成了黏膜薄膜,它表现为唾液和上皮成分的混合膜。金颗粒的分布表明薄膜表面并不均匀,薄膜厚度可达100纳米。这项工作表明了以非侵入性方式在人类颊上皮细胞上直接可视化和表征黏膜薄膜的可行性。