Combined protective effect of zinc oxide nanoparticles and melatonin on cyclophosphamide-induced toxicity in testicular histology and sperm parameters in adult Wistar rats

Background: Cyclophosphamide [CP] has been known as an anticancer drug with several side effects on various organs such as a male reproductive system that can cause infertility

Objective: To evaluate the possible combined effects of zinc oxide nanoparticles [nZno] and melatonin [Mel] on sperm parameters and histopathological changes of the testis in CP-treated rats

Materials and Methods: 42 adult male Wistar rats were divided into six groups. GI: control, GII: 60 mg/kg/wk CP, GIII and GIV, 10 mg/kg/wk Mel and 5mg/kg/wk nZno and GV: 5 mg/kg/wk nZno and 10 mg/kg/wk Mel were given 2 hr prior to CP injection, respectively, GVI: 5mg/kg/wk nZno and 10 mg/kg/wk Mel simultaneously. After 8 wk of treatment, rats were sacrificed and testis and epididymis were harvested for further evaluation

Results: The CP-treated group showed significant decreases in the body, testes and epididymis weights and sperm parameters [sperm count, viability, motility] with an increase abnormal sperms when compared with the control [p<0.001], as well as many histological alterations included decreased diameters of seminiferous tubules and Johnsen's Testicular Score [with degeneration, desquamation, multi-nucleated giant cell formation], whereas combined treatment [GV], showed more protective effects on CP-induced reproductive system damage compared with groups III or IV [p<0.001]

Conclusion: These results suggest simultaneous administration of Mel and nZno have more effectively protections against CP-induced reproductive damage than Mel or nZno alone

Expression of spermatogonial and pluripotency markers in spermatogonial stem cells after treatment with different culture factors

As condition and component of culture determine fate map of spermatogonial stem cells [SSCs], the aim of this study was to evaluate of growth factors GDNF, LIF and RA on proliferation and differentiation of SSC. SSCs were cultured in two groups: The first group GDNF and LIF and the second group RA. The number of clumps and colony formation was monitored during 1 month in culture. To identification of the colony, stained with PLZF using immunostaining. Pluripotency gene Oct 4 and neural markers MAP2, NeuroD and Nestin were analyzed by RT-PCR. In the presence of GDNF and LIF, cells proliferated rapidly and many compact clumps were appeared whereas after exposure to RA cells formed small clumps. The results of immunocytochemistry shows PLZF was detected in the group GDNF and LIF. RT-PCR showed high level expression Oct 4 in the group GDNF and LIF whereas neural markers MAP2, NeuroD and Nestin were expressed in the group RA. GDNF and LIF are essential for self-renewal and colony formation of SSCs that confirm the stem cells activity of these cells but RA inhibits stem cell activity of SSCs and induces neural differentiation of these

Effects of essential and non-essential amino acids on in-vitro maturation, fertilization and development of immature bovine oocytes

Addition of amino acids to the culture medium is beneficial for embryonic development in many species. The objective of this study was to investigate the effects of amino acids on the in vitro maturation and embryonic development of the bovine oocyte. Bovine ovaries were collected from a local abattoir and brought into laboratory. Cumulus-oocyte complexes [COCs; n=1212] were aspirated from follicles [2-8 mm in diameter] and randomly assigned to four groups for maturation in culture: [1] Basic medium alone as control; [2] Basic medium supplemented with 2% MEM essential amino acids solution; [3] Basic medium supplemented with 1% MEM non-essential amino acids solution; and [4] Basic medium supplemented with 2% MEM essential amino acids solution + 1% MEM non-essential amino acids solution. COCs were incubated in 1 ml maturation medium in an Organ culture dish at 38.5°C in an atmosphere of 5% CO2 with high humidity. After 24 h of culture, 372 oocytes were fixed to determine maturation rate and the remaining oocytes were used for in vitro fertilization [IVF]. Following 18 h of insemination, 437 oocytes were fixed and examined for fertilization and 403 oocytes were further cultured. There were no differences in maturation rates and penetration rates among the four groups. Although oocyte cleavage rates were not different in the four groups, embryo development up to the 8-cell stage and blastocyst were significantly higher [p<0.05] in Group [2] and [4] than in the Control and Group [3]. These results indicate that the presence of amino acids, especially essential amino acids in the maturation medium is beneficial to oocyte cytoplasmic maturation and subsequent early embryo development in vitro