School of Pharmacy, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, United Kingdom; Wellcome Centre Human Genetics, University of Oxford, Oxford, United Kingdom.
School of Pharmacy, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, United Kingdom.
Acta Biomater. 2022 Jan 15;138:285-300. doi: 10.1016/j.actbio.2021.11.016. Epub 2021 Nov 17.
microRNA-31 (miR-31) has been identified to be downregulated in pathologies associated with delayed wound repair. Thus, it was proposed that the delivery of a plasmid encoding miR-31 (pmiR-31) to the skin could hold potential in promoting wound healing. Effective delivery of pmiR-31 was potentiated by encapsulation with the CHAT peptide to form nanocomplexes, this improved cellular entry and elicited a potent increase in miR-31 expression in vitro in both skin human keratinocyte cell line (HaCaT) and human microvascular endothelial cell line (HMEC-1). Transfection efficiencies with CHAT/pEFGP-N1 were significant at 15.2 ± 8.1% in HMEC-1 cells and >40% in HaCaT cells. In this study, the CHAT/pmiR-31 nanocomplexes at a N:P ratio of 10 had an average particle size of 74.2 nm with a cationic zeta potential of 9.7 mV. Delivery of CHAT/pmiR-31 to HaCaT and HMEC-1 cells resulted in significant improvements in cell migration capacity and increased angiogenesis. In vivo studies were conducted in C57BL/6 J mice were CHAT/pmiR-31 was delivered via electrospun PVA nanofibres, demonstrating a significant increase in epidermal (increase of ∼38.2 µm) and stratum corneum (increase of 8.2 µm) layers compared to controls. Furthermore, treatment in vivo with CHAT/pmiR-31 increased angiogenesis in wounds compared to controls, with a significant increase in vessel diameter by ∼20.4 µm compared against a commercial dressing control (Durafiber™). Together, these data demonstrate that the delivery of CHAT/pmiR-31 nanocomplexes from electrospun PVA nanofibres represent an innovative therapy for wound repair, eliciting a positive therapeutic response across both stromal and epithelial tissue compartments of the skin. STATEMENT OF SIGNIFICANCE: This study advances research regarding the development of our unique electrospun nanofibre patch to deliver genetic nanoparticles into wounds in vivo to promote healing. The genetic nanoparticles are comprised of: (a) plasmid micro-RNA31 that has been shown to be downregulated in pathologies with delayed wound repair and (b) a 15 amino acid linear peptide termed CHAT. The CHAT facilitates complexation of miR-31 and cellular uptake. Herein, we report for the first time on the use of CHAT to deliver a therapeutic cargo pmiR-31 for wound healing applications from a nanofibre patch. Application of the nanofibre patch resulted in the controlled delivery of the CHAT/pmiR-31 nanoparticles with a significant increase in both epidermal and stratum corneum layers compared to untreated and commercial controls.
微小 RNA-31（miR-31）已被确定在与延迟伤口修复相关的病理中下调。因此，有人提出，将编码 miR-31 的质粒（pmiR-31）递送至皮肤可能具有促进伤口愈合的潜力。通过用 CHAT 肽包封来形成纳米复合物，有效地递送了 pmiR-31，这改善了细胞进入并在体外的皮肤人角质形成细胞系（HaCaT）和人微血管内皮细胞系（HMEC-1）中引起了 miR-31 表达的强烈增加。在 HMEC-1 细胞中，CHAT/pEFGP-N1 的转染效率为 15.2±8.1%，而在 HaCaT 细胞中>40%。在这项研究中，CHAT/pmiR-31 纳米复合物在 N：P 比为 10 时具有 74.2nm 的平均粒径和 9.7mV 的阳离子 ζ 电位。将 CHAT/pmiR-31 递送至 HaCaT 和 HMEC-1 细胞导致细胞迁移能力的显著改善和血管生成的增加。在 C57BL/6J 小鼠中进行了体内研究，通过电纺 PVA 纳米纤维递送至 CHAT/pmiR-31，与对照组相比，表皮（增加约 38.2μm）和角质层（增加 8.2μm）层显著增加。此外，与对照组相比，体内用 CHAT/pmiR-31 治疗增加了伤口中的血管生成，与商业敷料对照（Durafiber™）相比，血管直径增加了约 20.4μm。总的来说，这些数据表明，从电纺 PVA 纳米纤维递送电纺 CHAT/pmiR-31 纳米复合物代表了一种用于伤口修复的创新疗法，在皮肤的基质和上皮组织隔室中均引起了积极的治疗反应。研究意义：这项研究推进了我们独特的电纺纳米纤维贴片的开发研究，该贴片可将遗传纳米颗粒递送至体内伤口以促进愈合。遗传纳米颗粒由：（a）已显示在延迟伤口修复的病理中下调的质粒 micro-RNA31 和（b）称为 CHAT 的 15 个氨基酸线性肽组成。CHAT 促进 miR-31 的复合和细胞摄取。在这里，我们首次报告了使用 CHAT 从纳米纤维贴片递送电疗 cargopmiR-31 用于伤口愈合应用。纳米纤维贴片的应用导致 CHAT/pmiR-31 纳米颗粒的受控递送，与未处理和商业对照相比，表皮和角质层的厚度均显著增加。
