Department of Parasitology and Animal Diseases, Veterinary Research Institute, National Research Centre, Dokki, Giza, Egypt.
Department of Animal Reproduction and AI, Veterinary Research Institute, National Research Centre, El Buhouth Street, Dokki, Giza, Egypt.
Sci Rep. 2024 May 25;14(1):11957. doi: 10.1038/s41598-024-62015-0.
Hydatidosis causes a serious health hazard to humans and animals leading to significant economic and veterinary and public health concern worldwide. The present study aimed to evaluate the in vitro and ex vivo protoscolicidal effects of synthesized poly(amidoamine), PAMAM, nanoemulsion. In this study, PAMAM was characterized through dynamic light scattering technique to investigate the particle size and zeta potential of nanoemulsified polymer. For the in vitro and ex vivo assays, we used eosin dye exclusion test and scanning electron microscope (SEM) to evaluate the effects of the prepared and characterized PAMAM nanoemulsion against protoscoleces from Echinococcus granulosus sensu lato G6 (GenBank: OQ443068.1) isolated from livers of naturally infected camels. Various concentrations (0.5, 1, 1.5 and 2 mg/mL) of PAMAM nanoemulsion at different exposure times (5, 10, 20 and 30 min) were tested against protoscolices. Our findings showed that PAMAM nanoemulsion had considerable concentration- and time-dependent protoscolicidal effect at both in vitro and ex vivo experiments. Regarding in vitro assay, PAMAM nanoemulsion had a potent protoscolicidal effect when compared with the control group with a highest protoscolicidal activity observed at the concentration of 2 mg/mL at all exposure times, such that 100% of protoscolices were killed after 20 min of exposure. Also, the mortality of protoscolices was 100% after 30 min of exposure to 1 and 1.5 mg/mL of PAMAM nanoemulsion, in vitro. Concerning ex vivo assay PAMAM nanoemulsion recorded the highest mortality rates at the concentration of 2 mg/mL (55, 99.4 and 100% at 10, 20, 30 min, respectively). Ultrastructure examination of examined protoscolices after 20 min of exposure to PAMAM nanoemulsion showed a complete loss of rostellar hooks, disruption of suckers with disorganization of hooks with partial or complete loss of them, and damage of protoscolices tegument with loss of their integrity in the form of holes and contraction of the soma region were observed in 1.5 and 2 mg/mL of PAMAM, in vitro and ex vivo, showing more damage in the in vitro conditions. It can be concluded that PAMAM nanoemulsion is a promising protoscolicidal agent offering a high protoscolicidal effect at a short exposure time. Further in vivo studies and preclinical animal trials are required to evaluate its efficacy and clinical applications against hydatid cysts.
包虫病对人类和动物的健康造成严重危害，在全球范围内引起了重大的经济、兽医和公共卫生关注。本研究旨在评估合成的多（聚酰胺-胺），PAMAM，纳米乳剂的体外和体内杀原头节效果。在本研究中，通过动态光散射技术对 PAMAM 进行了表征，以研究纳米乳化聚合物的粒径和 Zeta 电位。对于体外和体内试验，我们使用伊红染料排除试验和扫描电子显微镜（SEM）来评估制备和表征的 PAMAM 纳米乳剂对源自天然感染骆驼肝脏的 Echinococcus granulosus sensu lato G6（GenBank：OQ443068.1）原头节的影响。测试了不同浓度（0.5、1、1.5 和 2 mg/mL）的 PAMAM 纳米乳剂在不同暴露时间（5、10、20 和 30 min）下对原头节的作用。我们的研究结果表明，PAMAM 纳米乳剂在体外和体内试验中均具有显著的浓度和时间依赖性杀原头节作用。在体外试验中，与对照组相比，PAMAM 纳米乳剂具有更强的杀原头节作用，在所有暴露时间下，浓度为 2 mg/mL 时观察到最强的杀原头节活性，暴露 20 min 后 100%的原头节被杀死。此外，1 和 1.5 mg/mL PAMAM 纳米乳剂暴露 30 min 后，原头节的死亡率为 100%，在体外。在体内试验中，PAMAM 纳米乳剂在浓度为 2 mg/mL 时记录到最高的死亡率（分别为 10、20 和 30 min 时的 55%、99.4%和 100%）。暴露于 PAMAM 纳米乳剂 20 min 后，对检查的原头节进行超微结构检查，结果显示完整的头棘丢失，吸盘破裂，吸盘上的棘钩排列紊乱，部分或完全丢失，原头节外被膜受损，形成孔，体区收缩，在 1.5 和 2 mg/mL 的 PAMAM 中观察到体外和体内，在体外条件下观察到更多的损伤。综上所述，PAMAM 纳米乳剂是一种很有前途的杀原头节剂，在短时间内具有很高的杀原头节作用。需要进一步的体内研究和临床前动物试验来评估其对包虫囊肿的疗效和临床应用。
