Therapeutic Potential of Different Injection Methods for Bone Marrow Mesenchymal Stem Cell Transplantation in Buslfan-Induced Male Rat Infertility

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Background: In recent years, bone marrow derived mesenchymal stem cells (BM-derived MSCs) have emerged as a powerful cell-based therapy for various diseases, including male infertility.Aim: Demonstrating the efficiency of BM-derived MSCs transplantation by different routes of injection to home and repair testis of busulfan-induced azoospermic rats. Material and methods:In the present study, rat BM-derived MSC was isolated and characterized for mesenchymal & hematopoietic markers using flow-cytometry.Induction of infertility was induced by two successive doses of 10 mg/kg of busulfan.Azoospermic rats were treated by BM-derived MSCs which were injected via various routes (IP, IV, and local in testis).After 60 days; sperm analyses were performed beside mainly Biochemical, histopathological, immunohistological, and ultrastructural investigations.Results: BM-derived MSCs were expressed by CD44+ve, CD105+ve, CD106+ve, CD73+ve, CD34-ve, and CD45-ve.Sperm analysis showed a substantial improvement in sperm morphology, motility, and count following treatment with BM-derived MSCs.Caspase-3 and PCNA immunoexpression accompanied with the levels of FSH, LH, testosterone, SOD, GSH and MDA depicted a considerable restoration of healthy levels after BM-derived MSCs treatment.The seminiferous tubules showed healthy morphology and spermatozoa were detected in their lumen according to the histopathological and ultrastructural analysis of BM-derived MSCs treated rats.Interestingly, BM-derived MSCs intravenous injection revealed the most significant infertility repair outcomes (P<0.05).Conclusion: Transplanted BM-derived MSCs had the potential to home in rat azoospermic testes and restore spermatogenesis.Consequently, the distinctive characteristics of BM-derived MSCs, such as their ability to differentiate and home, make them a promising cell-based therapeutic option for male infertility.regenerative medicine represents a new hope in treatment of many diseases including male factor infertility [8] .Recent researches have interested on the application of using the stem cell in the treatment of male infertility.Many studies have shown that embryonic stem cells (ESCs), and induced pluripotent stem cells (IPSCs) have the potentiality to differentiate into cells that exhibit certain germ cell markers.However, there are limitations in using ESCs and IPSCs, such as tumor propagation and moral concerns [9] .Also, early embryos have to be destroyed in order to isolate these ESCs [10] .On the other hand, spermatogonial stem cells (SSCs) are one of the most studied stem cells in treating male infertility, but they reside in testis with low population and there are difficulties in their isolation [11] .Mesenchymal stem/stromal cells (MSCs) have recently been proposed as a potential cell-based therapy for male infertility, whereas they do not have these issues in applications [12] .MSCs are common among researchers and physicians due to their

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