Centre des Sciences du Goût et de l'Alimentation, AgroSup Dijon, CNRS, INRA, Université de Bourgogne Franche-Comté , Dijon F-21000 , France.
ICB UMR CNRS 6303, Université de Bourgogne Franche-Comté , Dijon F-21078 , France.
Langmuir. 2019 Oct 1;35(39):12647-12655. doi: 10.1021/acs.langmuir.9b01979. Epub 2019 Sep 19.
The mucosal pellicle, also called salivary pellicle, is a thin biological layer made of salivary and epithelial constituents, lining oral mucosae. It contributes to their protection against microbiological, chemical, or mechanical insults. Pellicle formation depends on the cells' surface properties, and in turn the pellicle deeply modifies such properties. It has been reported that the expression of the transmembrane mucin MUC1 in oral epithelial cells improves the formation of the mucosal pellicle. Here, we describe an approach combining classical and functionalized tip atomic force microscopy and scanning microwave microscopy to characterize how MUC1 induces changes in buccal cells' morphology, hydrophobicity, and electric properties to elucidate the physicochemical mechanisms involved in the enhancement of the anchoring of salivary proteins. We show that MUC1 expression did not modify drastically the morphology of the epithelial cells' surface. MUC1 expression, however, resulted in the presence of more hydrophobic and more charged areas at the cell surface. The presence of salivary proteins decreased the highest attractive and repulsive forces recorded between the cell surface and a functionalized hydrophobic atomic force microscopy (AFM) tip, suggesting that the most hydrophobic and charged areas participate in the binding of salivary proteins. The cells' dielectric properties were altered by both MUC1 expression and the presence of a mucosal pellicle. We finally show that in the absence of MUC1, the pellicle appeared as a distinct layer poorly interacting with the cells' surface. This integrative AFM/scanning microwave microscopy approach may usefully describe the surface properties of various cell types, with relevance to the bioadhesion or biomimetics fields.
黏膜假膜,也称为唾液假膜,是由唾液和上皮成分组成的薄生物层,覆盖口腔黏膜。它有助于保护口腔免受微生物、化学或机械刺激。假膜的形成取决于细胞的表面性质,反过来,假膜又会深刻地改变这些性质。据报道,口腔上皮细胞跨膜粘蛋白 MUC1 的表达可改善黏膜假膜的形成。在这里,我们描述了一种结合经典和功能化尖端原子力显微镜和扫描微波显微镜的方法,以表征 MUC1 如何诱导颊细胞形态、疏水性和电特性的变化,从而阐明参与增强唾液蛋白锚定的物理化学机制。我们表明,MUC1 的表达并没有显著改变上皮细胞表面的形态。然而,MUC1 的表达导致细胞表面出现更多的疏水区和带电荷区。唾液蛋白的存在降低了细胞表面与功能化疏水性原子力显微镜 (AFM) 尖端之间记录到的最高吸引力和排斥力,表明最疏水区和带电荷区参与了唾液蛋白的结合。细胞的介电特性也受到 MUC1 表达和黏膜假膜存在的影响。最后,我们表明,在没有 MUC1 的情况下,假膜呈现出与细胞表面相互作用不良的明显层。这种整合的 AFM/扫描微波显微镜方法可用于描述各种细胞类型的表面特性,与生物黏附或仿生学领域相关。