Taudorf Elisabeth Hjardem, Lerche Catharina Margrethe, Vissing Anne-Cathrine, Philipsen Peter Alshede, Hannibal Jens, D'Alvise Janina, Hansen Steen Honore, Janfelt Christian, Paasch Uwe, Anderson Richard Rox, Haedersdal Merete
Bispebjerg University Hospital, University of Copenhagen, Department of Dermatology , Bispebjerg Bakke 23, DK-2400 Copenhagen NV , Denmark +45 35 31 60 04 ;
Expert Opin Drug Deliv. 2015 Jul;12(7):1059-69. doi: 10.1517/17425247.2015.1031216. Epub 2015 Apr 20.
Methotrexate (MTX) is a chemotherapeutic and anti-inflammatory drug that may cause systemic adverse effects. This study investigated kinetics and biodistribution of MTX delivered topically by ablative fractional laser (AFXL).
In vitro passive diffusion of 10 mg/ml MTX (1 w/v%) was measured from 0.25 to 24 h through AFXL-processed and intact porcine skin in Franz Cells (n = 46). A 2,940 nm fractional Erbium Yttrium Aluminium Garnet laser generated mid-dermal microchannels at 2.4% density, and 256 mJ/microchannel. HPLC quantified MTX-concentrations in extracts from mid-dermal skin sections, donor and receiver compartments. Fluorescence microscopy of UVC-activated MTX-fluorescence and desorption electro-spray ionization mass spectrometry imaging (DESI-MSI) evaluated MTX biodistribution.
AFXL-processed skin facilitated rapid MTX delivery through cone-shaped microchannels of 690 µm ablation depth, lined by the 47 µm thermal coagulation zone (CZ). Quantitatively, MTX was detectable by HPLC in mid-dermis after 15 min, significantly exceeded deposition in intact skin after 1.5 h, and saturated skin after 7 h at a 10-fold increased MTX-deposition versus intact skin (3.08 vs 0.30 mg/cm(3), p = 0.002). Transdermal permeation was < 1.5% of applied MTX before skin saturation, and increased up to 8.0% after 24 h. Qualitatively, MTX distributed into CZ within 15 min (p = 0.015) and further into surrounding dermal tissue after 1.5 h (p = 0.004). After skin saturation at 7 h, MTX fluorescence intensities in CZ and tissue were similar and DESI-MSI confirmed MTX biodistribution throughout the mid-dermal skin section.
MTX absorbs rapidly into mid-dermis of AFXL-processed skin with minimal transdermal permeation until skin saturation, suggesting a possible alternative to systemic MTX for some skin disorders.
甲氨蝶呤（MTX）是一种化疗和抗炎药物，可能会引起全身不良反应。本研究调查了通过剥脱性分数激光（AFXL）局部给药的MTX的动力学和生物分布。
在Franz扩散池（n = 46）中，测量了10 mg/ml MTX（1 w/v%）在0.25至24小时内通过AFXL处理的和完整的猪皮肤的体外被动扩散。一台2940 nm的分数钇铝石榴石激光以2.4%的密度和256 mJ/微通道产生真皮中层微通道。高效液相色谱法（HPLC）对真皮中层皮肤切片、供体和受体隔室提取物中的MTX浓度进行了定量分析。通过紫外线激活的MTX荧光显微镜和解吸电喷雾电离质谱成像（DESI-MSI）评估了MTX的生物分布。
AFXL处理的皮肤通过690 µm消融深度的锥形微通道促进了MTX的快速递送，该微通道由47 µm的热凝固区（CZ）衬里。定量分析显示，15分钟后在真皮中层可通过HPLC检测到MTX，1.5小时后显著超过完整皮肤中的沉积量，7小时后皮肤达到饱和，此时MTX的沉积量比完整皮肤增加了10倍（3.08对0.30 mg/cm³，p = 0.002）。在皮肤饱和前，经皮渗透量小于所施用MTX的1.5%，24小时后增加至8.0%。定性分析显示，MTX在15分钟内分布到CZ中（p = 0.015），1.5小时后进一步分布到周围的真皮组织中（p = 0.004）。7小时皮肤饱和后，CZ和组织中的MTX荧光强度相似，DESI-MSI证实MTX在整个真皮中层皮肤切片中均有生物分布。
MTX在皮肤饱和前能快速吸收进入AFXL处理皮肤的真皮中层，经皮渗透极少，这表明对于某些皮肤疾病，MTX局部给药可能是全身用药的一种替代方法。
