Department of Surgery, Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH 43210, USA; Division of Pharmaceutics and Pharmaceutical Chemistry, College of Pharmacy, The Ohio State University, Columbus, OH 43210, USA.
Department of Surgery, Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH 43210, USA; Center for Regenerative Medicine and Cell-Based Therapies, The Ohio State University, Columbus, OH 43210, USA.
Mol Ther. 2018 Sep 5;26(9):2178-2188. doi: 10.1016/j.ymthe.2018.04.021. Epub 2018 Apr 27.
Lyophilized keratinocyte-targeted nanocarriers (TLN) loaded with locked nucleic acid (LNA) modified anti-miR were developed for topical application to full thickness burn injury. TLN were designed to selectively deliver LNA-anti-miR-107 to keratinocytes using the peptide sequence ASKAIQVFLLAG. TLN employed DOTAP/DODAP combination pH-responsive lipid components to improve endosomal escape. To minimize interference of clearance by non-targeted cells, especially immune cells in the acute wound microenvironment, surface charge was neutralized. Lyophilization was performed to extend the shelf life of the lipid nanoparticles (LNPs). Encapsulation efficiency of anti-miR in lyophilized TLN was estimated to be 96.54%. Cargo stability of lyophilized TLN was tested. After 9 days of loading with anti-miR-210, TLN was effective in lowering abundance of the hypoxamiR miR-210 in keratinocytes challenged with hypoxia. Keratinocyte uptake of DiD-labeled TLN was selective and exceeded 90% within 4 hr. Topical application of hydrogel-dispersed lyophilized TLN encapsulating LNA anti-miR-107 twice a week significantly accelerated wound closure and restoration of skin barrier function. TLN application depleted miR-107 and upregulated dicer expression, which accelerated differentiation of keratinocytes. Expression of junctional proteins such as claudin-1, loricrin, filaggrin, ZO-1, and ZO-2 were significantly upregulated following TLN treatment. These LNPs are promising as topical therapeutic agents in the management of burn injury.
用于治疗全层烧伤的经冻干的角质形成细胞靶向纳米载体（TLN）负载了锁核酸（LNA）修饰的抗 miR。TLN 被设计为使用肽序列 ASKAIQVFLLAG 选择性地将 LNA-抗 miR-107 递送到角质形成细胞。TLN 采用 DOTAP/DODAP 组合 pH 响应脂质成分来改善内涵体逃逸。为了最大限度地减少非靶向细胞（特别是急性伤口微环境中的免疫细胞）清除的干扰，表面电荷被中和。进行冻干以延长脂质纳米颗粒（LNPs）的保质期。冻干 TLN 中抗 miR 的包封效率估计为 96.54%。测试了冻干 TLN 中 cargo 的稳定性。在用抗 miR-210 加载 9 天后，TLN 有效地降低了缺氧挑战的角质形成细胞中低氧 miR- miR-210 的丰度。DiD 标记的 TLN 被角质形成细胞摄取的选择性超过 90%，在 4 小时内即可达到。每周两次局部应用水凝胶分散的冻干 TLN 包封 LNA 抗 miR-107 显著加速了伤口闭合和皮肤屏障功能的恢复。TLN 应用使 miR-107 耗竭并上调了 dicer 的表达，从而加速了角质形成细胞的分化。TLN 处理后，连接蛋白如 Claudin-1、loricrin、filaggrin、ZO-1 和 ZO-2 的表达显著上调。这些 LNPs 有望成为治疗烧伤的局部治疗剂。
