الفهرس | Only 14 pages are availabe for public view |
Abstract Male causes of infertility constitute approximately half of infertility cases. Furthermore, cases of non-obstructive azoospermia (NOA) represent 10% of male infertility and have no definite effective treatment. Recently, stem cells have been suggested to treat different conditions. Adipose derived mesenchymal stem cells (ADMSCs) proved to be more convenient than bone marrow derived mesenchymal stem cells (BMMSCs). They could be extracted from different sources with minor invasive techniques, proliferate in less time and give more cell number. Animal models of NOA were induced by intratesticular CaCl2 injection. It causes irreversible chemical castration with minimal or no side effects. Accordingly, the current study was designed to evaluate the therapeutic role of (ADMSCs) in treatment of NOA induced by CaCl2. Forty five adult male albino rats of average weight of 225 gm were used in the current study. All rats were given oral paracetamol (1ml syrup) (before the injection). They were divided into four groups: group I: It included fifteen rats. Ten rats (subgroups Ia, Ib) were injected with a single intra-testicular dose of 0.1 ml saline/100gm body weight. Subgroup Ia: five rats were sacrificed after one week. Subgroup Ib: five rats were sacrificed after nine weeks. Subgroup Ic: five rats were sacrificed for the obtaining of adipose tissue derived mesenchymal stem cells (ADMSCs). group II: It included ten rats. Each rat was subjected to a single intra-testicular injection of calcium chloride (CaCl2) (2.5mg/testis/100gm body weight) dissolved in 0.1 ml saline/ testis. The animals were sacrificed after one week. group III: It included ten rats. They were given CaCl2 as in group II. After one week from CaCl2 injection, 100 𝜇L of ADMSCs mixture (106 cells) dissolved in 0.1 ml saline/ testis was injected intra-testicular. The animals were sacrificed eight weeks after stem cells injection. group IV: It included ten rats. Each rat was given CaCl2 in the same manner as in group II and sacrificed after nine weeks to be compared with group III. At the designated times, the animals were sacrificed by cervical dislocation. The testes were dissected out and processed for both light (H&E, PAS, Masson trichrome and Toludine blue stains) and electron microscopic examination. Sperm count smears sections preparation and total testosterone hormonal assay were done. Morphometric and statistical analysis were performed. The results of this study revealed spermatogenic epithelium degeneration with various degrees of apoptosis and necrosis in the seminiferous tubules of group II. Sloughing of epithelial lining of the seminiferous tubules with formation of necrotic tissue was detected. Damaged Leydig cells, inflammatory cells and multinucleated giant cells were also observed in group II. Injection of ADMSCs in group III resulted in great improvement of the structure of the testis being comparable to the control group. The seminiferous tubules showed well organized spermatogenic epithelium and sperms in the lumen. However, group IV showed a similar picture to that of group II with more damage of the testicular tissue and extensive collagen fibers deposition. Apoptosis and necrosis were obvious in this group with presence of damaged Leydig cells, chronic inflammatory cells (macrophages) in the interstitium. Statistically, groups II and IV showed a significant decrease in the mean sperm count and the serum testosterone level as compared to the control group. On the other hand, group III showed a non-significant change as compared to the control group. from the previous data, it was concluded that ADMSCs injection resulted in great improvement and regeneration of the structure of the testis following the testicular damage induced by NOA CaCl2 model. |