Occurrence of metachronous and synchronous secondary tumors in oral cavities has been associated with poor prognosis and decreased 5-year disease-free survival rates. The origin of secondary tumors in the oral cavity has been primarily attributed to cancer field effects (CFE) or malignancy-associated changes (MAC) in uninvolved areas. Classification of normal, cancerous and pre-cancerous oral lesions by in vivo Raman spectroscopy (RS) has already been demonstrated. In the present study, MAC/CFE in oral buccal mucosa were explored. In vivo Raman spectra from 84 subjects (722 spectra) under five categories - cancer and contralateral normal (opposite side of tumor), healthy controls (no tobacco habit, no cancer), habitués healthy controls (tobacco habit, no cancer) and non-habitués contralateral normal (no tobacco habit with cancer) were acquired. Mean and difference spectra suggest that loss of lipids and additional features representing proteins and DNA are characteristics of all pathological conditions, with respect to healthy controls. Spectral data were analyzed by PC-LDA followed by leave-one-out cross-validation. Results suggest that Raman characteristics of mucosa of healthy controls are exclusive, while those of habitués healthy controls are similar to those of contralateral normal mucosa. It was observed that the cluster of non-habitués contralateral normal mucosa is different from habitués healthy controls, suggesting that malignancy associated changes can be identified and also indicating that transformation of uninvolved oral mucosa due to tobacco habit or malignancy is different. The findings of the study demonstrate the potential of RS in identifying early transformation changes in oral mucosa and the efficacy of this approach in oral cancer applications.
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