Gomes Daré Regina, Beatriz Chieco Costa Ana, Silva Martins Tereza, Lopes Luciana B
Institute of Biomedical Sciences, University of São Paulo, 1524 Professor Lineu Prestes Avenue, 05508-000 São Paulo, SP, Brazil.
Institute of Biomedical Sciences, University of São Paulo, 1524 Professor Lineu Prestes Avenue, 05508-000 São Paulo, SP, Brazil.
Eur J Pharm Biopharm. 2024 Dec;205:114533. doi: 10.1016/j.ejpb.2024.114533. Epub 2024 Oct 15.
Chronic wounds represent a significant global health burden, characterized by delayed skin healing and associated comorbidities. The present study aimed to develop nanostructured lipid carriers (NLCs) as a topical delivery system for the co-administration of simvastatin and adenosine to address chronic wound management. The rationale behind the co-delivery approach was to mitigate the cytotoxicity associated with high-dose simvastatin, while preserving its therapeutic benefits through a potential synergistic or additive effect. A significant challenge in the development of these NLCs was the encapsulation of the highly hydrophilic adenosine within the hydrophobic lipid matrix. The NLCs were prepared using a hot homogenization-sonication method with a double emulsion technique and optimized through a series of formulation trials, employing various surfactants, solid and liquid lipids, to achieve efficient drug encapsulation, particularly for the hydrophilic adenosine. Optimized formulations F5- and F10-S/A 0.6 %/2 % (containing 0.6 % simvastatin and 2 % adenosine), exhibited promising physicochemical properties. The main difference was the liquid lipid used: F5 contained Miglyol 810 N, while F10 contained Capmul MCM C-8. Both formulations displayed a mean particle size below 230 nm, a polydispersity index (PDI) of approximately 0.2, and a zeta potential around -22 mV. While simvastatin association efficiency (AE) was nearly 100 %, adenosine AE was higher for F10 (24 %), compared to F5 (13.5 %). F5 demonstrated superior stability compared to F10, maintaining consistent particle size and PDI over a 60-day period. Formulation F5 also demonstrated superior cell-based in vitro performance compared to F10, with higher cell viability (MTT assay), greater cell proliferation induction (SRB assay), and enhanced cell proliferation and migration in the wound-scratch assay. While F10 displayed higher adenosine AE, F5 excelled in terms of stability and biological activity. The slight increase in intracellular reactive oxygen species levels observed with F5 may contribute to its enhanced proliferative effects. In-depth characterization revealed that F5 comprised spherical nanoparticles, and thermal analysis indicated no significant changes in the nanocarrier structure upon drug encapsulation. Additionally, ex vivo permeability study demonstrated superior skin retention of both simvastatin and adenosine for F5 compared to an emulsion control. Overall, the F5 nanocarrier demonstrated suitable physicochemical properties, cellular biocompatibility, induction of cell proliferation and migration events, and drug retention capacity in the skin layers, indicating its potential as a promising topical treatment for difficult-to-heal wounds.
慢性伤口是一项重大的全球健康负担，其特征为皮肤愈合延迟及相关合并症。本研究旨在开发纳米结构脂质载体（NLCs）作为一种局部给药系统，用于联合给予辛伐他汀和腺苷以解决慢性伤口管理问题。联合给药方法背后的基本原理是减轻与高剂量辛伐他汀相关的细胞毒性，同时通过潜在的协同或相加作用保留其治疗益处。开发这些NLCs的一个重大挑战是将高度亲水性的腺苷包裹在疏水性脂质基质中。使用热均质化 - 超声处理方法结合双乳液技术制备NLCs，并通过一系列配方试验进行优化，采用各种表面活性剂、固体和液体脂质，以实现高效的药物包裹，特别是对于亲水性的腺苷。优化后的配方F5 - 和F10 - S/A 0.6%/2%（含有0.6%辛伐他汀和2%腺苷）表现出良好的物理化学性质。主要区别在于使用的液体脂质：F5含有Miglyol 810 N，而F10含有Capmul MCM C - 8。两种配方的平均粒径均低于230 nm，多分散指数（PDI）约为0.2，zeta电位约为 - 22 mV。虽然辛伐他汀的包封率（AE）接近100%，但与F5（13.5%）相比，F10的腺苷AE更高（24%）。与F刷显示出更高的稳定性，在60天内保持一致的粒径和PDI。配方F5在基于细胞的体外性能方面也优于F10，具有更高的细胞活力（MTT法）、更强的细胞增殖诱导（SRB法）以及在伤口划痕试验中增强的细胞增殖和迁移能力。虽然F10显示出更高的腺苷AE，但F5在稳定性和生物活性方面表现出色。F5观察到的细胞内活性氧水平的轻微升高可能有助于其增强的增殖作用。深入表征显示F5由球形纳米颗粒组成，热分析表明药物包裹后纳米载体结构无显著变化。此外，体外透皮研究表明，与乳液对照相比，F5的辛伐他汀和腺苷在皮肤中的保留率更高。总体而言，F5纳米载体表现出合适的物理化学性质、细胞生物相容性、诱导细胞增殖和迁移事件以及在皮肤层中的药物保留能力，表明其作为难愈合伤口的有前景的局部治疗方法的潜力。
