Preparation of Organisms for Inoculation in Animal Model

Four to five colonies of standard *C. albicans* strains were selected and suspended in normal saline (2 mL) and mixed vigorously. The suspended cells were collected by centrifugation at 2500× *g* for 10 min. Thereafter, the pelleted cells were washed with phosphate buffered saline thrice and finally suspended in the buffer to obtain the final cell concentration of 3 <sup>×</sup> <sup>10</sup><sup>8</sup> colony-forming units (CFU)/mL.

Formation of Oral Candidiasis in the Rat Model and Evaluation of the Developed Formulation Preparation of Immunocompromised Model

The efficacy of the developed HA-FS-NTZ for the oral candidiasis model was evaluated in immunocompromised animal models following the protocol of Martinez and team [23]. The acclimatized animals were immunocompromised first by the consumption of dexamethasone with tetracycline. The initial dose of dexamethasone (0.5 mg/L) and tetracycline (1 g/L) was supplied to the animals via drinking water for 7 days, followed by an increase in the concentration of dexamethasone (1 mg/L) and a reduction in the concentration of tetracycline (0.1 g/L) 1 day before infecting the animals. The same dose of dexamethasone with tetracycline was continued until the end of the study.
