Effects of adipose derived mesenchymal stem cells and erythropoietin in testicular torsion-induced the germ cell injury in the adult albino rat

ABSTRACT
Background: The impairment of spermatogenesis due to a failure in the germ cell proliferation and differentiation is considering one of the major factors of male infertility which is a common complication due to ischemic injury of the testis. So far, even after surgical correction and orchiopexy, there is no an effective method to restore the spermatogenesis. Erythropoietin (EPO) and different types of stem cells were used separately to rescue the testis from this complication.
Aim of the work: To compare the separate and combined effects of erythropoietin and adipose-derived stem cells (AD-MSCs) in the rat testis after the torsion de-torsion (T/D) injury.
Material and methods: A total of sixty adult male albino rats weighing (12-week-old) 200-250 grams were used in this study. They were divided randomly into five groups (10 rats for each group), in addition to 10 rats used as a source for AD-MSCs. The groups were: Group I (Control group): which subdivided into a negative control and Sham operated rats), Group II(torsion detorsion (T/D) group): Torsion of the left testis by rotating the testis 270O in a clockwise direction for 2 hours, followed by detorsion in an anticlockwise direction and then fixed in position till the scarification after 6weeks, Group III (Erythropoietin-treated): The left testes of all rats were exposed to 720o torsion for 2 hours de-torsion and intra testicular injection of 3X106AD-MSCs suspended in 0.5 ml of DMEM as a vehicle. Group IV (MSCs-treated group): The left testes of all rats were exposed to 720o torsion for 2 hours, de-torsion and received a single intra venous injection of erythropoietin 3000 u/Kg. Group V: (MSCs/erythropoietin-treated): The left testes of all rats were exposed to 720o torsion for 2 hours, de-torsion and received a single intra venous injection of erythropoietin 3000 u/Kg in addition to intra testicular injection of 3X106 MSCs suspended in 0.5 ml of DMEM. After the end of the study, all rats from different groups were scarified and the left testicular tissue was obtained, weighed, examined grossly and prepared for a histopathological (a light and an electron microscope) and an immunohistochemical examination. As well, a quantitative assessment of the spermatogenesis was done statistically.
Results: Histopathological examinations showed a severe destruction of seminiferous tubules of the left testes with a failure of the sperm maturation. Also; recognizable abnormalities of the ultrastructure of Sertoli, Leydig cells and all the stages of spermatogenesis in Group II (T/D) were noticed. In addition, there was a significant decrease in the testicular weight and the number of sperms. Affection was also observed but to a lesser degree in Groups III &IV (EPO and AD-MSCs).The greatest amelioration of the tissue damage and the best histological improvement of spermatogenesis, and the testicular weight with a high statistically significance were seen in Group V (EPO and AD-MSC, respectively).
Conclusion: The administration of EPO and AD-MSCs has a great protective effect on the testis and the spermatogenesis. It may be used as a promising therapy after T/D to keep the fertility.