Therapeutic effects of adipose-derived stem cells-based microtissues on erectile dysfunction in streptozotocin-induced diabetic rats

This study aimed to explore the therapeutic effects of adipose-derived stem cells （ADSCs）-based microtissues （MTs） on erectile dysfunction （ED） in streptozotocin （STZ）-induced diabetic rats. Fifty-six 8-week-old Sprague-Dawley rats received intraperitoneal injection of STZ （60 mg kg-1）, and 8 weeks later, the determined diabetic rats randomly received intracavernous （IC） injection of phosphate buffer solution （PBS）, ADSCs, or MTs. Another eight normal rats equally got IC injection of PBS. MTs were generated with a hanging drop method, and the injected cells were tracked in ADSC- and MT-injected rats. Four weeks after the treatments, intracavernous pressure （ICP）, histopathological changes in corpus cavernosum （CC）, and functional proteins were measured. Rat cytokine antibody array was used to detect ADSCs or MTs lysate. The results showed that MTs expressed vascular endothelial growth factor （VEGF）, nerve growth factor （NGF）, and tumor necrosis factor-stimulated gene-6 （TSG-6）. MTs injection had a higher retention than ADSCs injection and MTs treatment improved ICP, neuronal nitric oxide synthase （nNOS） expression, smooth muscle, and endothelial contents in diabetic rats, ameliorated local inflammation in CC better. Thus, our findings demonstrate that IC injection of MTs improves erectile function and histopathological changes in STZ-induced diabetic rats and appears to be more promising than traditional ADSCs. The underlying mechanisms involve increased cell retention accompanied with neuroprotection and anti-inflammatory behaviors of the paracrine factors.

Proteomic analysis and miRNA profiling of human testicular endothelial cell-derived exosomes:the potential effects on spermatogenesis

Testicular endothelial cells have been found to play an important role in spermatogenesis and fertility,but their mechanism is obscure.Exosomes released by various cells are recognized as cell–cell communication mediators during the initiation and progression of many diseases.Therefore,the current study aimed to investigate the protein and miRNA components of human testicular endothelial cell-derived exosomes(HTEC-Exos)and to explore their potential effects on spermatogenesis.In this study,HTEC-Exos were first isolated by the ultracentrifugation method,and then identified by nanoparticle tracking analysis,transmission electron microscopy(TEM),and western blotting.The characteristics of HTEC-Exos were examined by liquid chromatography–mass spectrometry and microRNA(miRNA)chip analysis.Bioinformatics analysis was performed to explore the potential role of the exosomal content on spermatogenesis.A total of 945 proteins were identified,11 of which were closely related to spermatogenesis.A total of 2578 miRNAs were identified.Among them,30 miRNAs demonstrated potential associations with male reproductive disorders,such as azoospermia,and spermatogenesis disorders.In particular,11 out of these 30 miRNAs have been proven to be involved in spermatogenesis based on available evidence.This study provides a global view of the proteins and miRNAs from HTEC-Exos,suggesting that HTEC-Exos may function as potential effectors during the process of spermatogenesis.

Comparative study of intracavernous pressure and cavernous pathology after bilateral caver nous n erve crushing and resection in rats

This study aimed to compare the effects of bilateral cavernous nerve crushing(BCNC)and bilateral cavernous nerve resection(BCNR)on intracavernous pressure(ICP)and cavernous pathology in rats and to explore the optimal treatment time for the BCNC and BCNR models.Seventy-two male rats aged 12 weeks were randomly divided into three equal groups:Sham(both cavernous nerves exposed only),BCNC(BCN crushed for 2 min),and BCNR(5 mm of BCN resected).Erectile function was then measured at 1 week,3 weeks,and 5 weeks after nerve injury,and penile tissues were harvested for histological and molecular analyses by immunohistochemistry,immuno fluoresce nee,Western blot,and cytokine array.We found that erectile function parameters including the maximum,area,and slope of ICP/mean arterial pressure(MAP)significantly decreased after BCNR and BCNC at 1 week and 3 weeks.At 5 weeks,no significant differences were observed in ICP/MAP between the BCNC and Sham groups,whereas the ICP/MAP of the BCNR group remained significantly lower than that of the Sham group.After BCNC and BCNR,the amount of neuronal-nitric oxide synthase-positive fibers,smooth muscle cells,and endothelial cells decreased,whereas the amount of collage n III con tent increased.These pathological cha nges recovered over time,especially in the BCNC group.Our fin dings demonstrate that BCNC leads to acute and reversible erectile dysfunction,thus treatme nt time should be restricted to the first 3 weeks post-BCNC.In contrast,the self-healing ability of the BCNR model is poor,making it more suitable for long-term treatment research.