Stereological Evaluation of Induction of Azoospermia after The Using Busulfan in Albino Hamster Testes
Background: Busulfan is an alkylating agent, which inhibits cell dividing by sticking to one of the DNA strands. The aim of the present study was establishment and stereological evaluation of testes of azoospermic animal model using busulfan in albino hamster. Materials and Methods: Male adult albino hamsters were randomly assigned into 3 groups. The first group was injected one dose of busulfan (10 mg/kg, intraperitoneally, Busilvex®, France) and their testes were removed on day 35 post injection. The second group received two doses of busulfan with 21 days interval and their testes were removed on day 35 after the second injection. The testes of control group were removed without busulfan therapy. The histopathologic sections (5 µm thickness) were stained with hematoxylin- eosin. In 10 circular transverse sections of tubules, inner, outer and total diameters, calculated areas of the cellular and luminal regions and cross sectional area of the tubule, number of profiles of seminiferous tubules per unit area of testis, and numerical density of the seminiferous tubules using a systematic random scheme were measured. The testes were rated for its spermatogenic potential on a modified spermatogenic scale of 0 to 5. The data of stereological indices of seminiferous tubules were analyzed by one-way ANOVA and LSD post-hoc test (SPSS 11.5). The spermatogenesis index of seminiferous tubules was compared using Mann-Whitney U test. Results: Lumen, cellular and total diameter, luminal, cellular and cross sectional area, number of tubules per unit area of testis, numerical density of the tubules and spermatogenesis index in hamsters that injected two doses of busulfan were more than the hamsters in one dose busulfan injected and control groups (p<0.05). Conclusion: Two doses of busulfan injection with 21 days interval produced an appropriate animal model of induced azoospermia with comparable stereological indices of seminiferous tubules 35 days after the last injection in hamster.