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Background: Orange-red fluorescence in the follicle openings, induced by ultraviolet A light, originates from porphyrins, the metabolic products of Propionibacteria acnes. Purpose: To investigate the relationship of orange-red follicular fluorescence with the severity of acne and the amount of sebum secretion. Methods: Twenty-five volunteers were included. The severity of acne was rated on a 4-point scale. The casual sebum level was measured using a Sebumeter and the follicular fluorescence was determined using the camera Visiopor. Results: Casual sebum level and the intensity of fluorescence (percentage of the area and number of orange-red spots) were higher at the T zone than at the U zone in all patients regardless of their skin type. Sebum amount and area of fluorescence spots were significantly negative in correlation with the clinical grade of acne. There was a significant positive correlation between the orange-red fluorescence and the casual sebum level. Conclusion: The orange-red fluorescence showed stronger correlation with the presence of non-inflammatory acne lesions (comedones) and high sebum amount than the presence of inflammatory acne lesions (pustules) and low sebum amount. The fluorescence diagnostic imaging could be useful in the objective evaluation and monitoring of treatment efficacy in subjects with acne-prone skin and patients with acne.