Stromal changes in apparently normal mucosa of smokers and pan chewers - a multi-parametric approach

Aim: To compare and contrast the various changes in the connective tissue among patients with the habits of smoking, pan chewing and controls by using fluorescence spectroscopy, histopathology and transmission electron microscopy.
Method: Thirty subjects were categorized into three groups: pan chewers, smokers and controls without any oral lesions. Fluorescence spectroscopy was carried out using FLUOROMAX-2. Excitation spectroscopy was performed at 280 and 320nm respectively and Emission spectroscopy was performed at 340 and 390nm excitation. Subsequently, histopathological evaluation and transmission electron microscopy was done for biopsies taken from test groups and controls.
Results: The mean, standard deviation and test of significance of mean values between different groups for intensity-380nm, intensity-420nm and intensity - 460nm at 320 nm excitations, showed that the Mean values in group I and group II were significantly higher than the mean value in group III with a p-value of less than 0.001. Considering the histopathological parameters, the pattern of the sub-epithelial connective tissue, the presence of chronic inflammatory cells and lysis of connective tissue was significant. The ultrastructural analysis revealed the presence of epithelioid, spindle or elongated and stellate shaped fibroblasts in the connective tissue.
Conclusion: In this pilot study, we could find variations in emission characteristics of various amino acids which correlated with histopathology and electron microscopy. Our study suggests that there are connective tissue changes in oral mucosa among smokers and pan chewers, though it is apparently normal in clinical presentation. These initial connective tissue changes could determine the progression of altered mucosa to a pre-cancer or cancer, which is further related to other complex interactions. Elaborate studies are required to evaluate the significance of our hypothesis.