In vitro production of sperm is a desirable idea for fertility preservation in azoospermic men and prepubertal boys suffering from cancer. In this study, a biocompatible porous scaffold based on a triad mixture of silk fibroin (SF), alginate (Alg), and laminin (LM) is developed to facilitate the differentiation of mouse spermatogonia stem cells (SSCs). Following SF extraction, the content is analyzed by SDS-PAGE and stable porous 3D scaffolds are successfully prepared by merely Alg, SF, and a combination of Alg-SF, or Alg-SF-LM through freeze-drying. Then, the biomimetic scaffolds are characterized regarding the structural and biological properties, water absorption capacity, biocompatibility, biodegradability, and mechanical behavior. Neonatal mice testicular cells are seeded on three-dimensional scaffolds and their differentiation efficiency is evaluated using real-time PCR, flow cytometry, immunohistochemistry. Blend matrices showed uniform porous microstructures with interconnected networks, which maintained long-term stability and mechanical properties better than homogenous structures. Molecular analysis of the cells after 21 days of culture showed that the expression of differentiation-related proteins in cells that are developed in composite scaffolds is significantly higher than in other groups. The application of a composite system can lead to the differentiation of SSCs, paving the way for a novel infertility treatment landscape in the future.
Testes from nine male Malin x Santa-Ines rams with an average body weight of 43.1+/-3.53 kg, were used to study the effects of palm kernel cake (PKC) based diet on spermatogenic cells and to assess copper (Cu) levels in liver, testis and plasma in sheep. Animals were divided into three groups and randomly assigned three dietary treatments using restricted randomization of body weight in completely randomized design. The dietary treatments were 60% palm kernel cake plus 40% oil palm frond (PKC), 60% palm kernel cake plus 40% oil palm frond supplemented with 23 mg/kg dry matter of molybdenum as ammonium molybdate [(NH(4))(6)Mo(7)O(24).4H(2)O] and 600 mg/kg dry matter of sulphate as sodium sulphate [Na(2)SO(4)] (PKC-MS) and 60% concentrate of corn-soybean mix+40% oil palm frond (Control), the concentrate was mixed in a ratio of 79% corn, 20% soybean meal and 1% standard mineral mix. The results obtained showed that the number of spermatogonia, spermatocytes, spermatids and Leydig cells were not significantly different among the three treatment groups. However, spermatozoa, Sertoli cells and degenerated cells showed significant changes, which, may be probably due to the Cu content in PKC. Liver and testis Cu levels in the rams under PKC diet was found to be significantly higher (P<0.05) than rams in Control and PKC-MS diets. Plasma Cu levels showed a significant increase (P<0.05) at the end of the experiment as compared to at the beginning of the experiment for PKC and Control. In conclusion, spermatogenesis is normal in rams fed the diet without PKC and PKC supplemented with Mo and S. However spermatogenesis was altered in the PKC based diet probably due to the toxic effects of Cu and the significant changes in organs and plasma. Thus, Mo and S play a major role in reducing the accumulation of Cu in organs.
Introduction: We aim to investigate the effect of vasectomy on the histology of the testis as well as to evaluate DNA fragmentation in testicular tissue of male mice. Methods: Bilateral vasectomy was performed on 20 mature male mice; 10 control mice underwent sham-operation. After 6 weeks, the testes were evaluated for histological changes and DNA fragmentation by single cell gel electrophoresis (comet assay). Results: Marked alterations were observed in the testes of vasectomized mice, including degeneration of spermatids, thickened basement membrane, dilatation of the seminiferous tubules, exfoliation of germ cells, reduction in the seminiferous cell population, vacuolated appearance of the epithelium in the tubules and marked interstitial fibrosis. Single cell gel electrophoresis showed a highly significant (P
Effect of Centella asiatica aqueous leaf extract was assessed on spermatogenesis and sperm quality in mice. The experiment was conducted by giving three different doses of C. asiatica extract orally; 25mg/kg, 50mg/ kg and 100mg/kg body weight daily for four weeks respectively. Control group was treated with normal saline (0.9% NaCl). Marked decreases in sperm count and motility were found in sperm collected from the cauda epididymis of treated mice. The treatment, however did not affect sperm mortality and morphology. Histologically, signs of inhibition of testicular cells appeared clearly at dose 100mg/kg, when the lumen of seminiferous tubules were only full with spermatocytes and spermatids. The present results show that the plant has antispermatogenic effect in mice. It is therefore, suggested that leaf extracts of C. asiatica possess antifertility activity in male mice.
Kesan ekstrak berakua daun Centella asiatica telah dikaji ke atas proses spermatogenesis dan kualiti sperma mencit. Kajian dijalankan dengan memberikan tiga dos C. asiatica secara oral masing-masing; 25mg/kg, 50mg/ kg and 100mg/kg berat tubuh mencit setiap hari selama empat minggu. Kumpulan kawalan diberikan larutan salin normal (0.9% NaCl). Pengurangan dalam bilangan dan motiliti sperma yang ketara telah diperhatikan pada semua kumpulan mencit yang diberikan ekstrak C. asiatica. Walau bagaimanapun pemberian ekstrak C. asiatica tidak mendatangkan kesan terhadap mortaliti dan morfologi sperma. Kajian histologi pula menunjukkan terdapat kesan perencatan sel-sel spermatogenik testis pada dos C. asiatica 100mg/kg di mana lumen pada tubul seminiferus hanya dipenuhi dengan sel-sel spermatosit dan spermatid. Hasil kajian menunjukkan tumbuhan ini mempunyai kesan antispermatogenik pada mencit. Oleh itu dicadangkan bahawa ekstrak daun C. asiatica mempunyai aktiviti antikesuburan pada mencit jantan.