ECAUSE Toxoplasma gondii is a parasite with several developmental stages and strains, antiToxoplasma medications must be efficient for various stages and strains. The commonly used
drugs need long treatment periods and have hazardous side effects. Combining therapies may
decrease the pharmaceutical dosages while preserving therapeutic effects and minimizing toxicity.
This study evaluated the efficacy of combining spiramycin and metronidazole with
testosterone/progesterone and the combination loaded on Zeolitic imidazolate frameworks (ZIF-8)
nanoparticles in treating chronic toxoplasmosis in animal models. Mice were orally infected by brain
suspension containing Toxoplasma gondii cysts. They were sorted into female and male groups, and
each group was then divided into five subgroups: negative control (uninfected), positive control
(infected untreated), treated with spiramycin and metronidazole, treated with spiramycin and
metronidazole in combination with testosterone/progesterone, and treated with the combinationloaded ZIF-8 nanoparticles. Tested compounds' efficacy was determined by parasitological
assessment of parasite load in the brain, Toxoplasma DNA quantification in brain tissues using realtime PCR, and histopathological evaluation. Treatment of infected mice with spiramycin and
metronidazole in combination with testosterone or progesterone significantly decreased the parasite
burden. Moreover, mice treated with the drug combination carried on ZIF-8 showed a marked
decrease in parasitic load versus the other groups. All treated groups showed a reduction in
Toxoplasma DNA concentrations in brain tissues, with significant variation compared to positive
controls. Likewise, combo therapy successfully cured the histopathological alterations in the infected
mice's brains. In conclusion, sex hormones can modulate Toxoplasma infection, and their loading on
ZIF-8 nanoparticles increased anti-toxoplasmic effect.