Lee Woan-Ruoh, Lin Yin-Ku, Alalaiwe Ahmed, Wang Pei-Wen, Liu Pei-Yin, Fang Jia-You
Graduate Institute of Medical Sciences, Taipei Medical University, Taipei, Taiwan; Department of Dermatology, Taipei Medical University Shuang Ho Hospital, New Taipei City, Taiwan.
School of Traditional Chinese Medicine, Chang Gung University, Kweishan, Taoyuan, Taiwan; Department of Traditional Chinese Medicine, Chang Gung Memorial Hospital at Keelung, Keelung, Taiwan.
Mol Ther Nucleic Acids. 2020 Mar 6;19:240-251. doi: 10.1016/j.omtn.2019.11.013. Epub 2019 Nov 20.
The poor permeability of topically applied macromolecules such as small interfering RNA (siRNA) has inhibited the translation to clinical application. In this study, the fractional CO laser-assisted approach was developed to describe siRNA permeation enhancement mediated by the created microchannels for silencing the gene to treat psoriasiform lesions. In vitro permeation using Franz cell and in vivo interleukin (IL)-6 silencing using psoriasis-like plaque in mice were evaluated to verify the impact of the laser irradiation. Low-fluence laser exposure enabled a significant increase in skin transport of siRNA, peptide, and 5-fluorouracil (5-FU). The laser treatment resulted in the enhancement of siRNA flux by 33- and 14-fold as compared to the control in nude mouse and pig skin, respectively. The laser exposure also promoted siRNA penetration across psoriatic and photoaging skins with the deficient barrier, although the enhancement level was minor compared to that of intact skin. The 3D images of confocal microscopy revealed a diffusion of macromolecules into the laser-created microchannels; the radial and vertical distribution to the surrounding and deep tissues followed this. A single laser treatment and the following topical siRNA administration were able to reduce IL-6 expression by 64% in the psoriatic skin model. Laser assistance led to the marked improvement in the plaque and the reduction of specific cytokine expression, keratinocyte proliferation, and neutrophil infiltration. Our data support the use of the fractional laser for delivery of functional nucleic acid into the skin and the target cells.
局部应用的大分子（如小干扰RNA，siRNA）渗透性较差，这阻碍了其向临床应用的转化。在本研究中，开发了分数CO2激光辅助方法，以描述由所创建的微通道介导的siRNA渗透增强作用，从而沉默基因以治疗银屑病样病变。使用Franz扩散池进行体外渗透实验，并利用小鼠银屑病样斑块进行体内白细胞介素（IL）-6沉默实验，以验证激光照射的影响。低能量激光照射能够显著增加siRNA、肽和5-氟尿嘧啶（5-FU）的皮肤转运。与裸鼠和猪皮肤中的对照组相比，激光治疗分别使siRNA通量提高了33倍和14倍。激光照射还促进了siRNA穿过屏障功能不足的银屑病皮肤和光老化皮肤，尽管与完整皮肤相比，增强程度较小。共聚焦显微镜的三维图像显示大分子扩散到激光创建的微通道中；随后是向周围和深部组织的径向和垂直分布。单次激光治疗及随后的局部siRNA给药能够使银屑病皮肤模型中的IL-6表达降低64%。激光辅助导致斑块明显改善，特定细胞因子表达、角质形成细胞增殖和中性粒细胞浸润减少。我们的数据支持使用分数激光将功能性核酸递送至皮肤和靶细胞。
