Abstract：
Cutaneous leishmaniasis (CL) is a vector-borne disease characterized by skin lesions that can evolve into high-magnitude ulcerated lesions. Thus, this study aimed to develop an innovative nanoemulsion (NE) with clove oil, Poloxamer® 407, and multiple drugs, such as amphotericin B (AmB) and paromomycin (PM), for use in the topical treatment of CL.
METHODS: Droplet size, morphology, drug content, stability, in vitro release profile, in vitro cytotoxicity on RAW 264.7 macrophages, and antileishmanial activity using axenic amastigotes of Leishmania amazonensis were assessed for NEs.
RESULTS: After optimizing the formulation parameters, such as the concentration of clove oil and drugs, using an experimental design, it was possible to obtain a NE with an average droplet size of 40 nm and a polydispersion index of 0.3, and these parameters were maintained throughout the 365 days. Furthermore, the NE showed stability of AmB and PM content for 180 days under refrigeration (4 °C), presented a pH compatible with the skin, and released modified AmB and PM. NE showed the same toxicity as free AmB and higher toxicity than free PM against RAW 264.7 macrophages. The same activity as free AmB, and higher activity than free PM against amastigotes L. amazonensis.
CONCLUSIONS: It is possible to develop a NE for the treatment of CL; however, complementary studies regarding the antileishmanial activity of NE should be carried out.
METHODS: Droplet size, morphology, drug content, stability, in vitro release profile, in vitro cytotoxicity on RAW 264.7 macrophages, and antileishmanial activity using axenic amastigotes of Leishmania amazonensis were assessed for NEs.
RESULTS: After optimizing the formulation parameters, such as the concentration of clove oil and drugs, using an experimental design, it was possible to obtain a NE with an average droplet size of 40 nm and a polydispersion index of 0.3, and these parameters were maintained throughout the 365 days. Furthermore, the NE showed stability of AmB and PM content for 180 days under refrigeration (4 °C), presented a pH compatible with the skin, and released modified AmB and PM. NE showed the same toxicity as free AmB and higher toxicity than free PM against RAW 264.7 macrophages. The same activity as free AmB, and higher activity than free PM against amastigotes L. amazonensis.
CONCLUSIONS: It is possible to develop a NE for the treatment of CL; however, complementary studies regarding the antileishmanial activity of NE should be carried out.
摘要：
皮肤利什曼病（CL）是一种媒介传播的疾病，其特征是皮肤病变可以演变成高程度的溃疡病变。因此，这项研究旨在开发一种创新的纳米乳液（NE）与丁香油，泊洛沙姆®407和多种药物，如两性霉素B（AmB）和巴龙霉素（PM），用于CL的局部治疗。
方法：液滴大小，形态学,药物含量,稳定性,体外释放曲线，对RAW264.7巨噬细胞的体外细胞毒性，并评估了使用亚马逊利什曼原虫的轴突动物的抗利什曼原虫活性。
结果：优化配方参数后，例如丁香油和药物的浓度，使用实验设计，可以获得平均液滴尺寸为40nm且多分散指数为0.3的NE，并且这些参数在整个365天内都保持不变。此外，NE在冷藏（4°C）下显示了180天的AmB和PM含量的稳定性，呈现与皮肤相容的pH值，并发布了修改后的AmB和PM。NE对RAW264.7巨噬细胞表现出与游离AmB相同的毒性和比游离PM更高的毒性。与免费AmB相同的活动，对amastigotesL.amazonensis的活性高于游离PM。
结论：可以开发用于治疗CL的NE；但是，应进行有关NE的抗利什曼酶活性的补充研究。
方法：液滴大小，形态学,药物含量,稳定性,体外释放曲线，对RAW264.7巨噬细胞的体外细胞毒性，并评估了使用亚马逊利什曼原虫的轴突动物的抗利什曼原虫活性。
结果：优化配方参数后，例如丁香油和药物的浓度，使用实验设计，可以获得平均液滴尺寸为40nm且多分散指数为0.3的NE，并且这些参数在整个365天内都保持不变。此外，NE在冷藏（4°C）下显示了180天的AmB和PM含量的稳定性，呈现与皮肤相容的pH值，并发布了修改后的AmB和PM。NE对RAW264.7巨噬细胞表现出与游离AmB相同的毒性和比游离PM更高的毒性。与免费AmB相同的活动，对amastigotesL.amazonensis的活性高于游离PM。
结论：可以开发用于治疗CL的NE；但是，应进行有关NE的抗利什曼酶活性的补充研究。
