Ozbaş-Turan Suna, Akbuğa Jülide, Sezer Ali Demir
Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Marmara University, Istanbul, Turkey.
Oligonucleotides. 2010 Jun;20(3):147-53. doi: 10.1089/oli.2009.0222.
Skin delivery of antisense oligonucleotides (AsODNs) has exciting potential in the treatment of skin diseases. However, the therapeutic applications of oligonucleotide-based therapies are limited by the instability of these molecules toward nucleases, short half-life in vivo, and insufficient cellular uptake. The purpose of this study was to investigate in vivo antisense effect of AsODN-loaded chitosan nanoparticles after topical application. AsODN-loaded chitosan nanoparticles were topically applied to Sprague Dawley rats (adult and baby). At 1, 3, 6, 9, and 12 days posttransfection, animals' skin samples were taken for measurement of beta-galactosidase (beta-Gal) expression and histological control. After topical application of AsODN-loaded chitosan nanoparticles in different doses, beta-Gal expression reduced significantly. Highest inhibition was observed after 6 days of transfection of nanoparticles. Free AsODNs exhibited 35% of beta-Gal inhibition on the first day. beta-Gal expression was inhibited in approximately 82-85% with transfection of nanoparticles containing 30 microg AsODNs at 6 days. The antisense effect of AsODN-loaded chitosan nanoparticle in baby skin was evaluated at 6 days: 77-86% of beta-Gal suppression was measured and differences between the doses were not significant. Thus, chitosan nanoparticles are useful carrier for delivery of AsODNs into skin cells of rats and may be used for topical application on human skin.
反义寡核苷酸（AsODNs）经皮肤给药在皮肤病治疗方面具有令人兴奋的潜力。然而，基于寡核苷酸的疗法的治疗应用受到这些分子对核酸酶的不稳定性、体内半衰期短以及细胞摄取不足的限制。本研究的目的是研究局部应用负载AsODN的壳聚糖纳米颗粒后的体内反义效应。将负载AsODN的壳聚糖纳米颗粒局部应用于成年和幼年的Sprague Dawley大鼠。在转染后1、3、6、9和12天，采集动物的皮肤样本以测量β-半乳糖苷酶（β-Gal）表达并进行组织学对照。局部应用不同剂量的负载AsODN的壳聚糖纳米颗粒后，β-Gal表达显著降低。在纳米颗粒转染6天后观察到最高抑制率。游离AsODNs在第一天表现出35%的β-Gal抑制率。在6天时，含有30μg AsODNs的纳米颗粒转染后，β-Gal表达被抑制约82 - 85%。在6天时评估了负载AsODN的壳聚糖纳米颗粒在幼鼠皮肤中的反义效应：测量到β-Gal抑制率为77 - 86%，且不同剂量之间的差异不显著。因此，壳聚糖纳米颗粒是将AsODNs递送至大鼠皮肤细胞的有用载体，并且可用于人类皮肤的局部应用。
