Observation On Nanocrystallization Albendazole Against Echinoeococcus Gransosus Protoscolices

ObjectiveThis study is aimed at comparing the parasiticidal efficacy of different concentration of Albendazole powder and nanocrystallization Albendazole against protoscolices of Echinococcus granulosus in vitro,in order to improve the chemotherapeutic efficacy to provide experimental bases.MethodsProtoscolices of Echinococcus granulosus were collected aseptically from the liver hydatid cysts of naturally infected sheep and cultured in 6-well plates with RPMI 1640.The protoscolices were co-cultured with nanocrystallization Albendazole and Albendazole powder at concentrations of: 3.13,6.25,12.5,25?g/ml in vitro,and set up the blank control group.Evidence of viability and changes in morphology was observed by an inverted microscope after 0.1% eosin staining everyday.The changes of ultrastructure in protoscolices were observed by scanning electron microscopy(SEM).The protoscolices were collected at 24 and48 hours for the activity of caspase-3 was quantified with a caspase-3 activity assay kit.ELISA were used to characterize the levels of NQO-1 and HO-1 in the protoscolices cultured with different concentration of Albendazole.ResultsAlbendazole powder and nanocrystallization Albendazole demonstrate a different extent of suppressive effect on the growth of E.g protoscolices in vitro.The results of inverted microscope,significant difference of E.g protoscolices mortality was found among the with drugs at various concentrations compared to control group(P<0.05),and a significant difference between Albendazole powder and nanocrystallization Albendazole(P<0.05).Obvious inhibitory effect was regarded with Albendazole at concentrations of 12.5?g/ml and25?g/ml.After 7 days post-incubation with 12.5,25 ?g/ml nanocrystallization Albendazole,the viability of the protoscolices had decreased to 27.4% in 12.5?g/ml,and viability was approximately 11.4% in 25?g/ml.The percentage of protoscolices death increased with time.SEM showed that the significant site of drug destruction was the tegument region of the protoscolices.The ultra structural alterations including contraction of the soma surface,rostella distortion,and shedding of hooks.The presence of irregular cavities in the tegument of the protoscolices were visualized cultured with a higher concentration of Albendazole powder and nanocrystallization Albendazole.Nevertheless,control group almost no change in their morphology and ultra structure was observed throughout SEM.The caspase-3 activity increased in protoscolices treated with different concentration of Albendazole for 24 h and 48 h compared with control group.Albendazole powder and nanocrystallization Albendazole efficiently inhibited the levels of NQO-1and HO-1 in the protoscolices in a time-and dose-dependent manner(P<0.05).Nanocrystallization Albendazole displays a stronger effect than Albendazole powder.ConclusionAlbendazole powder and nanocrystallization Albendazole demonstrate a different extent of suppressive effect on the growth of E.g protoscolices in vitro,and a significant difference between Albendazole powder and nanocrystallization Albendazole,in sight to be a new dosage form of the drug to treat echinococcosis.As exposure time goes on,the lethal effect of Albendazole anti-E.g protoscolices in vitro was more distinct.