Topical delivery of anti-sense oligonucleotides using low-frequency sonophoresis
Topical delivery of anti-sense oligonucleotides using low-frequency sonophoresis
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DOI：
10.1007/s11095-004-7674-6
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发表时间：
2004-12-01
影响因子：
3.7
通讯作者：
Mitragotri, S
中科院分区：
文献类型：
--
作者：
Tezel, A;Dokka, S;Mitragotri, S
目的.寡核苷酸的局部递送虽然对于治疗皮肤病有吸引力，但受限于角质层的低渗透性。在此，我们评估了低频超声（20 kHz，2.4 W/cm（2））在将治疗上显著量的反义寡核苷酸递送到皮肤中的潜力。使用放射性标记的寡核苷酸在体外定量寡核苷酸渗透的皮肤渗透。超声和ODN作用10分钟后，皮肤表层中寡核苷酸（ODN）的估计浓度范围为供体浓度的0.5%至5%。使用荧光标记的寡核苷酸和磺酰罗丹明B的显微镜评价揭示了向皮肤的异质渗透。异质性渗透导致局部转运途径（LTP）的形成，其占据总暴露皮肤面积的约5%。使用与抗体特异性反应的寡核苷酸的免疫组织化学研究也证实了ODNs渗透到LTP中。组织学研究表明，没有引起皮肤由于超声应用的总体结构变化。这些结果显示使用低频超声成功递送反义寡核苷酸。
Purpose. Topical delivery of oligonucleotides, though attractive for the treatment of skin disorders, is limited by the low permeability of the stratum corneum. Herein, we assessed the potential of low-frequency ultrasound (20 kHz, 2.4 W/cm(2)) in delivering therapeutically significant quantities of anti-sense oligonucleotides into skin.Methods. Dermal penetration of oligonucleotides penetration was quantified in vitro using radiolabeled oligonucleotides.Results. Estimated concentrations of oligonucleotides (ODNs) in the superficial layers of the skin ranged from similar to0.5% to 5% of the donor concentration after a 10-min application of ultrasound and ODN. Microscopic evaluations using fluorescently labeled oligonucleotides and sulforhodamine B revealed heterogeneous penetration into the skin. Heterogenous penetration led to the formation of localized transport pathways (LTPs), which occupied about 5% of the total exposed skin area. Immuno-histochemical studies using an oligonucleotide that reacts specifically with an antibody also confirmed penetration of ODNs into LTPs. Histologic studies revealed that no gross structural changes were induced in the skin due to ultrasound application.Conclusions. These results show successful delivery of anti-sense oligonucleotides using low-frequency ultrasound.