de Araujo Margarete Moreno, Borgheti-Cardoso Livia Neves, Praça Fabíola Garcia, Marcato Priscyla Daniely, Bentley Maria Vitória Lopes Badra
School of Pharmaceutical Sciences of Ribeirao Preto, University of São Paulo, Av. do Café, s/n, Ribeirão Preto 14040-903, SP, Brazil.
J Funct Biomater. 2023 Jul 17;14(7):374. doi: 10.3390/jfb14070374.
Small interfering RNA (siRNA) molecules have limited transfection efficiency and stability, necessitating the use of delivery systems to be effective in gene knockdown therapies. In this regard, lipid-polymeric nanocarriers have emerged as a promising class of nanoparticles for siRNA delivery, particularly for topical applications. We proposed the use of solid lipid-polymer hybrid nanoparticles (SLPHNs) as topical delivery systems for siRNA. This approach was evaluated by assessing the ability of SLPHNs-siRNA complexes to internalize siRNA molecules and both to penetrate skin layers in vitro and induce gene knocking down in a skin cell line. The SLPHNs were formed by a specific composition of solid lipids, a surfactant polymer as a dispersive agent, and a cationic polymer as a complexing agent for siRNA. The optimized nanocarriers exhibited a spherical shape with a smooth surface. The average diameter of the nanoparticles was found to be 200 nm, and the zeta potential was measured to be +20 mV. Furthermore, these nanocarriers demonstrated excellent stability when stored at 4 °C over a period of 90 days. In vitro and in vivo permeation studies showed that SLPHNs increased the cutaneous penetration of fluorescent-labeled siRNA, which reached deeper skin layers. Efficacy studies were conducted on keratinocytes and fibroblasts, showing that SLPHNs maintained cell viability and high cellular uptake. Furthermore, SLPHNs complexed with siRNA against Firefly luciferase (siLuc) reduced luciferase expression, proving the efficacy of this nanocarrier in providing adequate intracellular release of siRNA for silencing specific genes. Based on these results, the developed carriers are promising siRNA delivery systems for skin disease therapy.
小分子干扰RNA（siRNA）分子的转染效率和稳定性有限，因此需要使用递送系统才能在基因敲低疗法中发挥有效作用。在这方面，脂质-聚合物纳米载体已成为一类有前景的用于siRNA递送的纳米颗粒，特别是对于局部应用。我们提出使用固体脂质-聚合物杂化纳米颗粒（SLPHNs）作为siRNA的局部递送系统。通过评估SLPHNs-siRNA复合物内化siRNA分子的能力，以及在体外穿透皮肤层和在皮肤细胞系中诱导基因敲低的能力，对该方法进行了评估。SLPHNs由特定组成的固体脂质、作为分散剂的表面活性剂聚合物和作为siRNA络合剂的阳离子聚合物形成。优化后的纳米载体呈球形，表面光滑。发现纳米颗粒的平均直径为200 nm，zeta电位测量为+20 mV。此外，这些纳米载体在4°C下储存90天期间表现出优异的稳定性。体外和体内渗透研究表明，SLPHNs增加了荧光标记的siRNA的皮肤渗透，其可到达更深的皮肤层。对角质形成细胞和成纤维细胞进行了功效研究，结果表明SLPHNs保持了细胞活力并具有高细胞摄取率。此外，与针对萤火虫荧光素酶的siRNA（siLuc）复合的SLPHNs降低了荧光素酶的表达，证明了这种纳米载体在为沉默特定基因提供足够的细胞内siRNA释放方面的功效。基于这些结果，所开发的载体是用于皮肤病治疗的有前景的siRNA递送系统。
