Topical Lyophilized Targeted Lipid Nanoparticles in the Restoration of Skin Barrier Function following Burn Wound
Topical Lyophilized Targeted Lipid Nanoparticles in the Restoration of Skin Barrier Function following Burn Wound
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DOI：
10.1016/j.ymthe.2018.04.021
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发表时间：
2018-09-05
影响因子：
12.4
通讯作者：
Sen, Chandan K.
中科院分区：
文献类型：
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作者：
Li, Jilong;Ghatak, Subhadip;Sen, Chandan K.
研制了冷冻干燥角质形成细胞靶向纳米载体(TLN-kappa)，负载锁定核酸(LNA)修饰的抗miR，用于烧伤创面的局部应用。TLN kappa被设计用来使用多肽序列ASKAIQVFLLAG选择性地将LNA-anti-miR-107递送到角质形成细胞。TLN kappa使用DOTAP/DODAP结合pH响应性脂质成分来改善内体逃逸。为了最大限度地减少非靶向细胞，特别是急性创面微环境中免疫细胞对清除的干扰，表面电荷被中和。为了延长脂质纳米粒(LNPs)的货架期，进行了冷冻干燥。冻干TLN-kappa中抗miR的包封率为96.54%。对TLN kappa冻干后的货物稳定性进行了测试。负载抗miR-210抗体9天后，TLN-kappa可有效降低缺氧条件下角质形成细胞低氧受体miR-210的表达。角质形成细胞对DiD标记的TLN kappa有选择性摄取，4小时内摄取率达90%以上。局部应用水凝胶分散的冻干化TLN-kappa包被LNA抗miR-107，每周两次，可显著加快创面闭合和皮肤屏障功能的恢复。TLN-kappa/抗miR-107抑制miR-107的表达，上调DICER的表达，从而促进角质形成细胞的分化。经TLN kappa/anti-miR-107处理后，Claudin-1、loricrin、filaggrin、ZO-1和ZO-2等连接蛋白的表达显著上调。这些LNPs有望在烧伤治疗中作为局部治疗药物。
Lyophilized keratinocyte-targeted nanocarriers (TLN kappa) loaded with locked nucleic acid (LNA) modified anti-miR were developed for topical application to full thickness burn injury. TLN kappa were designed to selectively deliver LNA-anti-miR-107 to keratinocytes using the peptide sequence ASKAIQVFLLAG. TLN kappa 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 kappa was estimated to be 96.54%. Cargo stability of lyophilized TLN kappa was tested. After 9 days of loading with anti-miR-210, TLN kappa was effective in lowering abundance of the hypoxamiR miR-210 in keratinocytes challenged with hypoxia. Keratinocyte uptake of DiDlabeled TLN kappa was selective and exceeded 90% within 4 hr. Topical application of hydrogel-dispersed lyophilized TLN kappa encapsulating LNA anti-miR-107 twice a week significantly accelerated wound closure and restoration of skin barrier function. TLN kappa/anti-miR-107 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 kappa/anti-miR-107 treatment. These LNPs are promising as topical therapeutic agents in the management of burn injury.