Understanding retention and metabolization of aroma compounds using an in vitro model of oral mucosa.

The mechanism leading to aroma persistence during eating is not fully described. This study aims at better understanding the role of the oral mucosa in this phenomenon. Release of 14 volatile compounds from different chemical classes was studied after exposure to in vitro models of oral mucosa, at equilibrium by Gas-Chromatography-Flame Ionization Detection (GC-FID) and in dynamic conditions by Proton Transfer Reaction- Mass Spectrometry (PTR-MS). Measurements at equilibrium showed that mucosal hydration reduced the release of only two compounds, pentan-2-one and linalool (p < 0.05), and suggested that cells could metabolize aroma compounds from different chemical families (penta-2,3-dione, trans-2-hexen-1-al, ethyl hexanoate, nonan- and decan-2-one). Dynamic analyses for pentan-2-one and octan-2-one evidenced that the constituents of the mucosal pellicle influenced release kinetics differently depending on molecule hydrophobicity. This work suggests that mucosal cells can metabolize aroma compounds and that non-covalent interactions occur between aroma compounds and oral mucosa depending on aroma chemical structure.

Nanoscale Mapping of the Physical Surface Properties of Human Buccal Cells and Changes Induced by Saliva.

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.

Mechanisms of astringency: Structural alteration of the oral mucosal pellicle by dietary tannins and protective effect of bPRPs.

The interaction of tannins with salivary proteins is involved in astringency. This paper focussed on saliva lining oral mucosae, the mucosal pellicle. Using a cell-based model, the impact of two dietary tannins (EgC and EgCG) on the mucosal pellicle structure and properties was investigated by microscopic techniques. The role of basic Proline-Rich-Proteins (bPRPs) in protecting the mucosal pellicle was also evaluated. At low (0.05 mM) tannin concentration, below the sensory detection threshold, the distribution of salivary mucins MUC5B on cells remained unaffected. At 0.5 and 1 mM, MUC5B-tannin aggregates were observed and their size increased with tannin concentration and with galloylation. In addition, 3 mM EgCG resulted in higher friction forces measured by AFM. In presence of bPRPs, the size distribution of aggregates was greatly modified and tended to resemble that of the "no tannin" condition, highlighting that bPRPs have a protective effect against the structural alteration induced by dietary tannins.

The role of saliva in aroma release and perception.

Aroma perception is an important factor driving food acceptance. Volatile organic compounds (VOCs) are released from the food matrix and then reach the receptors located in the nasal cavity, leading to their perception. These steps are closely dependent on the physicochemical properties of the volatile compounds and the food matrix, but also on human physiology. Among the different physiological parameters involved, the literature reports that saliva has various effects on VOCs and therefore appears as a major actor impacting the perception of aroma. This article reviews how saliva takes part in aroma release, considering both in vitro and in vivo approaches, and how it may affect perception. It describes the direct mechanisms (molecular interactions, enzymatic conversion, salting-out effect, dilution) involving salivary components (salts, proteins including enzymes, microbiota) that can modify the release of aroma compounds. It also considers the indirect impact of saliva, such as changes of aroma diffusion through modification of the physicochemical properties of the food matrix.

The membrane-associated MUC1 improves adhesion of salivary MUC5B on buccal cells. Application to development of an in vitro cellular model of oral epithelium.

OBJECTIVES: The mucosal pellicle is a thin layer of salivary proteins, mostly MUC5B mucins, anchored to epithelial oral cells. This pellicle is involved in protection of oral mucosae against abrasion, pathogenic microorganisms or chemical xenobiotics. The present study aimed at studying the involvement of MUC1 in mucosal pellicle formation and more specifically in salivary MUC5B binding using a cell-based model of oral epithelium. DESIGN: MUC1 mRNAs were not detected in TR146 cells, and therefore a stable cell line named TR146/MUC1 expressing this protein was developed by transfection. TR146 and TR146/MUC1 were incubated with human saliva in order to evaluate retention of MUC5B by epithelial cells. RESULTS: The cell surface of both TR146 and TR146/MUC1 was typical of a squamous non-keratinized epithelium, with the presence of numerous microplicae. After incubation for 2h with saliva diluted in culture medium (1:1) and two washes with PBS, saliva deposits on cells appeared as a loose filamentous thin network. MUC5B fluorescent immunostaining evidenced a heterogeneous lining of confluent cell cultures by this salivary mucin but with higher fluorescence on TR146/MUC1 cells. Semi-quantification of MUC5B bound to cells confirmed a better retention by TR146/MUC1, evaluated by Dot Blot (+34.1%, p<0.05) or by immunocytochemistry (+44%, p<0.001). CONCLUSION: The membrane-bound mucin MUC1 is a factor enhancing the formation of the mucosal pellicle by increasing the binding of salivary MUC5B to oral epithelial cells. An in vitro model suitable to study specifically the function and properties of the mucosal pellicle is proposed.