Enzyme and Protein Delivery to Pellicle-Coated Reconstituted Human Oral Epithelium

Objectives: To measure and visualise the delivery of enzymes and proteins from a toothpaste to a pellicle coated reconstituted human oral epithelium
Methods: Reconstituted human oral epithelium was pellicle coated by treating with saliva for 2 hours at 37°C and then rinsing with water. Delivery was measured by treating the reconstituted human oral epithelium with an aqueous slurry of a toothpaste containing enzymes and proteins, rinsing with water after each treatment. Bound enzymes and proteins were then eluted from the surface of the epithelium using an SDS solution and submitted to mass spectrometric analysis. Delivery was also visualised by treating the pellicle coated reconstituted human oral epithelium with toothpaste containing fluorescently labelled enzymes and proteins, rinsing after each treatment with water. Samples were then formalin fixed and paraffin embedded, transversely sectioned and imaged using fluorescence microscopy.
Results: Delivery of the enzymes and proteins from the toothpaste could be detected using mass spectrometric analysis and showed a clear contrast with the control sample, a non-enzyme and protein containing toothpaste. Delivery of fluorescently-labelled enzymes and proteins to the upper surface of the epithelium was also successfully visualised using fluorescence microscopy.
Conclusions: We have demonstrated delivery of enzymes and proteins in vitro from a toothpaste to a pellicle coated reconstituted oral epithelium.