Trypanosoma cruzi trypomastigotes and amastigotes in vitro viability after incubation with benznidazole -loaded polymeric nanoparticles

MURACA, GIULIANA; MARIA PIÑEIRO; MATIAS MARCHETTO; CHAIN, CECILIA YAMIL; CISNEROS, JOSÉ SEBASTIÁN; SOTO, CATALINA ALBA; ISLAN, GERMÁN A.; TALEVI, ALAN

La Plata

Congreso; Sociedad Argentina de Protozoología; 2023

SAP

Trypanosoma cruzi is the causative agent of Chagas disease, an endemic infection that affects approximately 6-7 million people worldwide, mostly in Latin America. The only two available drugs, nifurtimox and benznidazole, are ineffective against the chronic stage of the disease. Intracellular persistent dormant parasites are hypothesized as a factor of resistance to pharmacotherapy. Therefore, pharmaceutical formulations capable of enhancing distribution may result in improved therapeutic output. The production of nano-carriers is a novel strategy that aims to modify pharmacokinetics parameters. Our goal was to develop polymeric carriers (EU) loaded with benznidazole (BNZ) to evaluate their effect on the trypomastigotes and amastigotes forms of the parasites. EU were prepared by the emulsification by ultrasonication technique. Formulations were characterized in terms of entrapment efficiency (%EE), size, TEM, polydispersity index, Z-potential, and in vitro antitrypanosomal activity. Formulations showed high %EE (74%), spherical shape, an average size of 157 nm, low polydispersity (PDI 0.026) and Z-pot around -33 mV. In vitro antitrypanosomal assays were performed against the K98 T. cruzi clone. Trypomastigotes (1x105 per well) were cocultured with different dilutions of BNZ and EU-BNZ in RPMI medium in a 96 well-plate at 37 ºC in 5% CO2 for 24 h, then live parasites were counted in a Neubauer chamber to determine the parasite viability. For the evaluation against amastigotes, Vero cells were previously infected with trypomastigotes at MOI 1:1 then seeded in RPMI adding at least 2x104 cells per well. Freshly dilutions of BNZ and EU-BNZ were added. After 72 h, the cells were harvested and processed for flow cytometry analysis. Evaluation of EU against trypomastigotes and amastigotes showed both a similar effect as using free drug. In conclusion, EU-BNZ with suitable size and shape were developed, and antitrypanosomal activity resulted in a similar effect of free and nanoparticulated drug on trypomastigotes and amastigotes.