The use of digitized endoscopic imaging of 5‐ALA‐induced PPIX fluorescence to detect and diagnose oral premalignant and malignant lesions in vivo

5-aminolevulinic acid (ALA)-induced protoporphyrin IX (PPIX) fluorescence has shown an outstanding sensitivity for the assessment of oral lesions, but its application was hampered by low specificity due to the high false-positive rates. The purpose of our study was to explore the feasibility of quantifying PPIX fluorescence images to improve the diagnostic specificity for detecting early oral lesions in vivo. A digitized 5-ALA-mediated endoscopic imaging system was utilized to acquire PPIX fluorescence images from in vivo oral tissues. Forty-nine patients (118 biopsies) with known or suspected premalignant or malignant oral lesions were recruited for ALA-PPIX fluorescence endoscopic imaging. The red and blue channels of PPIX fluorescence images were digitized and stored for fluorescence quantification. The red-to-blue intensity ratios were calculated from the fluorescence images to correlate with histologic findings of the biopsies. The results showed that normal oral mucosa exhibited blue color of the back-scattered excitation light in the fluorescence images, whereas the suspicious lesions displayed bright reddish fluorescence. Applying the red-to-blue intensity ratio (I(R)/I(B)) as a diagnostic algorithm yielded a sensitivity of 92% and 98%, and specificity of 96% and 96%, for separating benign tissue from dysplasia, and cancer tissue, respectively, and a sensitivity and specificity of 98% and 92%, respectively, for differentiating cancer tissue from dysplasia in the oral cavity. Our study demonstrates that quantifying ALA-PPIX fluorescence endoscopic images associated with the red-to-blue intensity ratio as a diagnostic algorithm can provide good differentiation between the different stages of oral premalignancy and malignancy (p<0.0001, unpaired 2-sided Student's t-test), and thus has a potential to significantly improve the noninvasive diagnosis and evaluation of early oral neoplasia in vivo.