The effect of benznidazole-loaded nanostructured lipid carriers on Trypanosoma cruzi trypomastigotes and amastigotes

G. MURACA; M. L. SBARAGLINI; C. Y. CHAIN; G. R. CASTRO; G. A. ISLÁN; C. ALBA SOTO; V. ALVAREZ; A. TALEVI

Mendoza

Congreso; XI Congreso de la Sociedad Argentina de Protozoología; 2022

Chagas disease is an infection caused by Trypanosoma cruzi that affects approximately 8 millionpeople. Possibly due to the cellular internalization of the parasite after evasion of the hostimmune response, pharmacotherapy fails to eradicate the parasites in tissue reservoirs andcauses a low cure rate in chronic patients. Drug delivery in nanotechnological vehicles is a novelstrategy that aims to modify the pharmacokinetics parameters such as bioavailability andtargeted biodistribution to specific cell types and tissues. Our goal was to develop ananostructured lipid carrier (NLC) loaded with benznidazole (BNZ) to evaluate its effect on thecirculating and intracellular forms of the parasite. NLC were prepared by the hot-emulsificationand ultrasonication technique. Formulations were characterized in terms of entrapmentefficiency (%EE), particle size, polydispersity index, Z-potential, thermal and structuralproperties, and antitrypanosomal activity. Formulations showed high %EE (80%), sphericalshape, an average size of 165 nm, low polydispersity (PDI 0.3) and Z-pot around -5 mV.Antitrypanosomal assays were performed against the K98 T. cruzi clone. Trypomastigotes (7x104per well) were cocultured with different dilutions of BNZ and NLC-BNZ in RPMI 1640 medium ina 96 well-plate at 37 ºC for 24 h. The number of parasites alive after the incubation was countedusing an optical microscope. For the evaluation against amastigotes, Vero cells previouslyinfected with trypomastigotes at MOI 1:2 were seeded in RPMI at 2x104 cells per well. Freshlydilutions of BNZ and NLC-BNZ were added. After 72 h, the cells were harvested and processedfor flow cytometry analysis. Evaluation of the NLC-BNZ against trypomastigotes and amastigotesshowed a similar effect as using BNZ. In conclusion, NLC-BNZ with suitable size and shape weredeveloped, and antitrypanosomal activity resulted in a similar effect of the free drug and NLCBNZon trypomastigotes and amastigotes.