Dipartimento di Morfologia Umana e Scienze Biomediche - Città Studi, Italy.
Inhal Toxicol. 2012 May;24(6):382-9. doi: 10.3109/08958378.2012.679367.
Human oral mucosa is the combustion chamber of cigarette, but scanty evidence is available about the early smoke effects.
The present work aimed at evaluating from a morphological point of view whole smoke early effects on epithelial intercellular adhesion and keratinocyte terminal differentiation in a three-dimensional model of human oral mucosa.
Biopsies of keratinized oral mucosa of healthy nonsmoking women (n = 5) were collected. After culturing in a Transwell system, one fragment of each biopsy was exposed to the smoke of one single cigarette; the remnant represented the internal control. The distribution of epithelial differentiation markers (keratin-10, K10, and keratin-14, K14, for suprabasal and basal cells respectively), desmosomes (desmoglein-1, desmoglein-3), tight junctions (occludin), adherens junctions (E-cadherin, β-catenin), and apoptotic cells (p53, caspase 3) were evaluated by immunofluorescence.
Quantitative analysis of K14 immunolabeling revealed an overexpression in the suprabasal layers as early as 3 h after smoke exposure, without impairment of the epithelial junctional apparatus and apoptosis induction.
These results suggested that the first significant response to cigarette smoke came from the basal and suprabasal layers of the human oral epithelium. The considered model maintained the three-dimensional arrangement of the human mucosa in the oral cavity and mimicked the inhalation/exhalation cycle during the exposure to cigarette smoke, offering a good possibility to extrapolate the reported observations to humans.
人体口腔黏膜是香烟的燃烧室，但关于早期烟雾影响的证据很少。
本研究旨在从形态学角度评估整个烟雾对三维人口腔黏膜模型中上皮细胞间黏附及角蛋白细胞终末分化的早期影响。
收集 5 例健康非吸烟女性的角化口腔黏膜活检标本。在 Transwell 系统中培养后，每个活检标本的一个片段暴露于一支香烟的烟雾中；剩余部分为内部对照。通过免疫荧光法评估上皮分化标志物（角蛋白-10，K10 和角蛋白-14，K14，分别用于基底层和上层细胞）、桥粒（桥粒蛋白-1、桥粒蛋白-3）、紧密连接（occludin）、黏附连接（E-钙黏蛋白、β-连环蛋白）和凋亡细胞（p53、caspase 3）的分布。
K14 免疫标记的定量分析显示，暴露于烟雾后 3 小时，上层细胞中出现过表达，上皮连接装置未受损且未诱导细胞凋亡。
这些结果表明，香烟烟雾的第一个显著反应来自人口腔上皮的基底层和上层。所考虑的模型保持了人黏膜在口腔中的三维排列，并模拟了暴露于香烟烟雾时的吸入/呼出循环，为将报告的观察结果外推至人类提供了很好的可能性。
