The BALB/c mice were kept in a standard living situation with 12 : 12-hour dark-light cycle, 40–50% humidity, at 25 ± 3°C temperature. Free access to a standard diet and water were also provided. Twenty-eight female BALB/c mice (4 weeks old, 18 ± 4 g weight) were obtained from Pasteur Institute, Tehran, Iran. The infection in BALB/c mice was induced by using the amastigote form of L. major (strain reference no. MRHO/IR/75/ER). To do that, 0.1 mL of a suspension, containing 4000–5000 amastigotes (counted by a hemocytometer counting-chamber), was injected subcutaneously into the top of mice's tail-base, using insulin syringes. Lesions were appeared after about 3 weeks of inoculation. Mice were divided into four groups (n = 7); group E1 treated with the 5% concentration of Tabashir gel daily; group E2 received 10% gel daily; group C1 received normal saline every day, and group C2 treated by vehicle gel (2%, CMC) daily. Gel administrations were started topically from the day that open wounds were appeared and continued daily until the end of the third week. The size of the lesions was measured every three days, using vernier caliper. To do this, the length of the minor and major axes of the lesion at the base of the mouse's tail perpendicular to each other was measured and the area of the lesion was calculated in square millimeters using ellipse area formula.

At the end of the treatment, the animals were euthanized using a high dose of ether. A circular skin sample with 1 cm of the grossly healthy margin around the lesion was excised from the wound's site and used for histomorphometric assessments.

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