Preclinical investigation of cavernous nerve injury-induced erectile dysfunction treated with HAFSC-derived secretome

BACKGROUND: Erectile dysfunction (ED) significantly affects both psychological and physiological well-being, with limited effective treatments available for nerve injury-induced ED. Stem cell therapy, particularly using human amniotic fluid stem cells (hAFSCs), shows promise due to the secretome (hAFSC-S), which is rich in growth factors and chemoattractants. This study investigated the paracrine effects of hAFSC-S on erectile function recovery in a rat model of bilateral cavernous nerve crush (BCNC) injury. MATERIALS AND METHODS: Thirty rats were divided into the sham and BCNC groups. The BCNC group received 200 µL of hAFSC-S via intracavernous injection. Western blotting was used to evaluate endothelial markers, while both Western blotting and enzyme-linked immunosorbent assay were used to characterize the secretome. Four weeks post-treatment, intracavernous pressure (ICP) was measured to assess erectile function. Histological analysis, immunofluorescence staining, and electron microscopy were conducted to examine penile tissue and nerve integrity. RESULTS: The hAFSC-S-treated group demonstrated significantly higher ICP (91.77 ± 5.21 cmH 2 O) compared to the untreated BCNC group (66.60 ± 3.64 cmH 2 O). Histology revealed improved muscle integrity in the corpus cavernosum, increased smooth muscle content, and enhanced nerve myelination. Immunofluorescence staining showed elevated expression of neuronal markers, including nNOS and β-III-tubulin, in the major pelvic ganglion (0.42 ± 0.01%) and dorsal penile nerve (0.46 ± 0.01%). No significant differences were observed in vWF expression between the BCNC and BCNC + secretome groups in some regions (0.07 ± 0.009%). However, Western blot analysis revealed restored expression of endothelial markers CD31 and eNOS in penile tissue. CONCLUSION: hAFSC-S therapy enhanced nerve regeneration and penile tissue restoration, leading to improved erectile function in a BCNC rat model. These findings highlight its potential as a therapeutic option for ED.