Development of a Novel Ex-vivo 3D Model to Screen Amoebicidal Activity on Infected Tissue

Abstract

Amoebiasis is a parasitic disease that causes thousands of deaths every year, its adverse effects and resistance to conventional treatments have led to the search of new treatment options, as well as the development of novel screening methods. In this work, we implemented a 3D model of intestine and liver slices from hamsters that were infected ex vivo with virulent E . histolytica trophozoites. Results show preserved histology in both uninfected tissues as well as ulcerations, destruction of the epithelial cells, and inflammatory reaction in intestine slices and formation of micro abscesses, and the presence of amoebae in the sinusoidal spaces and in the interior of central veins in liver slices. The three chemically synthetized compounds T-001, T-011, and T-016, which act as amoebicides in vitro , were active in both infected tissues, as they decreased the number of trophozoites, and provoked death by disintegration of the amoeba, similar to metronidazole. However, compound T-011 induced signs of cytotoxicity to liver slices. Our results suggest that ex vivo cultures of precision-cut intestinal and liver slices represent a reliable 3D approach to evaluate novel amoebicidal compounds, and to simultaneously detect their toxicity, while reducing the number of experimental animals commonly required by other model systems.

Pharmaceutical activity of a synthetic heterocyclic (C₁₅H₁₂N₅OCl) compound on <i>Entamoeba histolytica</i> and <i>Giardia lamblia</i>

“…This result is similar to the one observed with metronidazole [28], especially at a concentration of 1 mg/kg. Also, this compound had a similar effect to metronidazole on Entamoeba cultures [15]. Thus, we believe this compound could be an excellent therapeutic alternative for the treatment of intestinal parasites.…”

“…When tested against E. histolytica and G. intestinalis, the synthetic substance 5-(3-chlorophenyl)-1-methyl-4-nitro-1H-imidazole demonstrated a high fatal dosage of 1.47 μM/mL [27]. Additional synthetic inhibitors were discovered to show potential chemotherapeutic activities comparable to metronidazole against trophozoites and/or cysts of G. intestinalis and E. histolytica [8,15,28].…”

“…Several synthetic compounds (5-(3-chlorophenyl)-1-methyl-4-nitro-1 H-imidazole, synthesized inhibitors of the Hsp90 class, and many other compounds) have been reported to have potential chemotherapeutic properties against E. histolytica and G. intestinalis [8,27]. Thus, some of the compounds tested as alternatives to metronidazole have had encouraging results on intestinal tract parasites [15,28].…”

Modeling Paratuberculosis in Laboratory Animals, Cells, or Tissues: A Focus on Their Applications for Pathogenesis, Diagnosis, Vaccines, and Therapy Studies

“…The protocols for culturing ruminant organoids from different digestive segments have been set out in [112][113][114][115][116][117][118][119][120]. Since the first report in 2009, intestinal organoids have evolved as a potential alternative to in vivo models for various experimental purposes, such as imaging, molecular analysis and gene editing, but also as reductionist approaches with which to study the interaction of epithelial cells with the other relevant actors in a physiological context, such as the immune cells [121,122] or microbiota [123,124], or even to evaluate the early host-pathogen interaction in the context of many infectious diseases (protozoa [125,126], virus [127], and bacteria [128][129][130][131][132][133]).…”