Trypanosoma cruzi trypomastigotes and amastigotes in vitro viability after incubation with benznidazoleloaded polymeric nanoparticles

MURACA GIULIANA; PIÑERO, MARÍA BELÉN; MARCHETTO, MATIAS ILDEBRANDO; CECILIA Y. CHAIN; CISNEROS, JOSÉ SEBASTIÁN; CATALINA ALBA SOTO; GERMÁN A. ISLAN; ALAN TALEVI

Encuentro; XXXIV Reunión Anual de la Sociedad Argentina de Protozoología; 2023

Sociedad Argentina de Protozoología

Trypanosoma cruzi is the causative agent of Chagas disease, an endemic infection thataffects approximately 6-7 million people worldwide. The only two available drugs,nifurtimox and benznidazole (BNZ), are ineffective against the chronic stage of thedisease. Intracellular persistent dormant parasites are hypothesized as a factor ofresistance to pharmacotherapy. Therefore, pharmaceutical formulations capable ofenhancing distribution may result in improved therapeutic. The production of nano-carriers isa novel strategy that aims to modify pharmacokinetics parameters. Our goal was to develop polymeric carriers (EU) loaded with BNZ to evaluate their effect on trypomastigotes and amastigotes. 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 vitroantitrypanosomal 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 mediumin a 96 well-plate at 37 ºC in 5% CO2 for 24 h, then live parasites were counted in a Neubauerchamber to determine the parasite viability.For the evaluation against amastigotes, Verocells 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 andamastigotes 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.