Posaconazole (POS) is an inhibitor of
ergosterol biosynthesis in clinical use for treating
invasive fungal infections. POS has potent and selective anti-Trypanosoma cruzi activity and has been evaluated as a possible treatment for
Chagas disease. Microtissues are a 3D culture system that has been shown to reproduce better tissue architecture and functionality than cell cultures in monolayer (2D). It has been used to evaluate chemotropic response as in vitro disease models. We previously developed an in vitro model that reproduces aspects of cardiac
fibrosis observed in
Chagas cardiomyopathy, using microtissues formed by primary cardiac cells infected by the T. cruzi, here called T. cruzi fibrotic cardiac microtissue (TCFCM). We also showed that the treatment of TCFCM with a TGF-β pathway inhibitor reduces
fibrosis. Here, we aimed to evaluate the effect of POS in TCFCM, observing parasite load and molecules involved in
fibrosis. To choose the concentration of POS to be used in TCFCM we first performed experiments in a monolayer of primary cardiac cell cultures and, based on the results, TCFCM was treated with 5 nM of POS for 96 h, starting at 144 h post-
infection. Our previous studies showed that at this time the TCFCM had established
fibrosis, resulting from T. cruzi
infection. Treatment with POS of TCFCM reduced 50 % of parasite load as observed by real-time PCR and reduced markedly the
fibrosis as observed by western blot and immunofluorescence, associated with a strong reduction in the expression of
fibronectin and
laminin (45 % and 54 %, respectively). POS treatment also changed the expression of
proteins involved in the regulation of
extracellular matrix proteins (TGF-β and TIMP-4, increased by 50 % and decreased by 58 %, respectively) in TCFCM. In conclusion, POS presented a potent trypanocidal effect both in 2D and in TCFCM, and the reduction of the parasite load was associated with a reduction of
fibrosis in the absence of external immunological effectors.