Lee Woan-Ruoh, Shen Shing-Chuan, Liu Ching-Ru, Fang Chia-Lang, Hu Chung-Hong, Fang Jia-You
Department of Dermatology, Taipei Medical University Hospital, Taipei, Taiwan.
J Control Release. 2006 Oct 27;115(3):344-53. doi: 10.1016/j.jconrel.2006.08.012. Epub 2006 Aug 23.
Topical delivery of antisense oligonucleotides (ASOs) and DNA is attractive for treatment of skin disorders. However, this delivery method is limited by the low permeability of the stratum corneum (SC). The objective of this study was to enhance and optimize the skin absorption of gene-based drugs by an erbium:yttrium-aluminum-garnet (Er:YAG) laser. The animal model utilized nude mice. In the in vitro permeation study, the Er:YAG laser treatment produced a 3-30-fold increase in ASO permeation which was dependent on the laser fluence and ASO molecular mass used. The fluorescence microscopic images showed a more-significant localization of a 15-mer ASO in the epidermis and hair follicles after laser application as compared with the control. The expressions of reporter genes coding for beta-galactosidase and green fluorescent protein (GFP) in skin were assessed by X-gal staining and confocal laser scanning microscopy. The SC ablation effect and photomechanical waves produced by the Er:YAG laser resulted in DNA expression being extensively distributed from the epidermis to the subcutis. The GFP expression in 1.4 J/cm2-treated skin was 160-fold higher than that in intact skin. This non-invasive, well-controlled technique of using an Er:YAG laser for gene therapy provides an efficient strategy to deliver ASOs and DNA via the skin.
反义寡核苷酸（ASO）和DNA的局部递送对于皮肤疾病的治疗具有吸引力。然而，这种递送方法受到角质层（SC）低渗透性的限制。本研究的目的是通过铒：钇铝石榴石（Er：YAG）激光增强和优化基于基因的药物的皮肤吸收。动物模型使用裸鼠。在体外渗透研究中，Er：YAG激光治疗使ASO渗透增加了3至30倍，这取决于所使用的激光能量密度和ASO分子量。荧光显微镜图像显示，与对照相比，激光照射后15聚体ASO在表皮和毛囊中的定位更明显。通过X-gal染色和共聚焦激光扫描显微镜评估皮肤中编码β-半乳糖苷酶和绿色荧光蛋白（GFP）的报告基因的表达。Er：YAG激光产生的SC消融效应和光机械波导致DNA表达从表皮广泛分布到皮下组织。1.4 J/cm2处理的皮肤中的GFP表达比完整皮肤中的高160倍。这种使用Er：YAG激光进行基因治疗的非侵入性、可控性良好的技术为通过皮肤递送ASO和DNA提供了一种有效的策略。
