Native fluorescence spectroscopic characterization of DMBA induced carcinogenesis in mice skin for the early detection of tissue transformation

Abstract

The objective of the study is to characterize the endogenous porphyrin fluorescence in a dimethylbenz(a)anthracene (DMBA) induced mouse skin tumor model using native fluorescence emission and excitation spectroscopy. Two intensity ratio parameters I₅₈₀/I₆₃₅ and I₄₂₀/I₅₁₅ were selected to represent the key fluorophore of endogenous porphyrins from emission and excitation spectra recorded in vivo from 31 DMBA treated animals and 5 control animals. In the emission spectrum, the endogenous porphyrin was elevated at 635 nm in different transformation lesions such as hyperplasia, papilloma, dysplasia, ESCC and WDSCC. This is corroborated by the endogenous porphyrin elevation at 420, 515, 550 and 588 nm in the WDSCC lesions from the excitation spectra. The elevation of endogenous porphyrin, probably protoporphyrin IX (PpIX), is due to biochemical and metabolic alterations in epithelial cells during tissue transformation. The loss of ferrochelatase activity might be responsible for enhanced PpIX in the transformed tissues. The sensitivity and specificity were determined for different lesion pairs from the scatter plot based on the discrimination value by validation with histopathological results. The emission intensity ratio I₅₈₀/I₆₃₅ at 405 nm excitation was selected to discriminate normal from hyperplasia, hyperplasia from papilloma, papilloma from dysplasia, dysplasia from early squamous cell carcinoma (ESCC), and ESCC from well differentiated squamous cell carcinoma (WDSCC) with specificities of 100%, 88%, 100%, 86%, and 100% and sensitivities of 100%, 80%, 100%, 100% and 100% respectively. Similarly, the excitation intensity ratio I₄₂₀/I₅₁₅ for 635 nm emission used to discriminate between WDSCC lesions and normal tissue gives 100% specificity and 100% sensitivity.