The aim of this study was to examine the in vitro skin delivery and in vivo protoporphyrin IX (PpIX) accumulation of topically applied 5-aminolevulinic acid (ALA) enhanced by a fractional laser pretreatment. This was achieved by applying an array of microscopic treatment zones (MTZ) to the skin by ablation of superficial stratum corneum in a determined area. Re-epithelialization determined by transepidermal water loss was completed within 1day after fractional laser irradiation. The conventional laser used in comparison showed more severe skin disruption and a greater recovery duration of 2days. The in vitro ALA permeation was measured using a Franz cell apparatus, with nude mouse skin and porcine skin as the permeation barriers. The efficacy of the enhancement was determined as a function of various laser fluences (2 and 3J/cm2) and number of passes (1-6 passes). The flux of ALA via laser-treated nude mouse skin was 27-124-fold higher than that across intact skin. A 3-260-fold increase in ALA flux was detected by using the porcine skin as the permeation barrier. The skin permeation was also investigated in a model of hyperproliferative skin obtained by repeated tape stripping. The results showed that the hyperproliferative skin was more permeable to ALA in comparison to the normal skin. The in vivo localization of PpIX in nude mouse skin was imaged using confocal laser scanning microscopy. As expected, an intense red fluorescence was observed in the lower epidermis and upper dermis after fractional laser irradiation. The penetration depth was also increased by the laser. The safety and efficacy of enhancing ALA permeation were demonstrated by using the fractional laser at low fluences.
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