Pericyte-Derived Extracellular Vesicle–Mimetic Nanovesicles Restore Erectile Function by Enhancing Neurovascular Regeneration in a Mouse Model of Cavernous Nerve Injury

Abstract

Background Extracellular vesicle (EV)–mimetic nanovesicles (NVs) from embryonic stem cells have been observed to stimulate neurovascular regeneration in the streptozotocin-induced diabetic mouse. Pericytes play important roles in maintaining penile erection, yet no previous studies have explored the effects of pericyte-derived NVs (PC-NVs) in neurovascular regeneration in the context of erectile dysfunction. Aim To investigate the potential effect of PC-NVs in neurovascular regeneration. Methods PC-NVs were isolated from mouse cavernous pericytes, and neurovascular regeneration was evaluated in an in vitro study. Twelve-week-old C57BL/6J mice were used to prepare cavernous nerve injury model. Erectile function evaluation, histologic examination of the penis, and Western blots were assessed 2 weeks after model creation and PC-NVs treatment. Outcomes The main outcomes of this study are PC-NVs characterization, intracavernous pressure, neurovascular regeneration in the penis, and in vitro functional evaluation. Results The PC-NVs were extracted and characterized by cryotransmission electron microscopy and EV-positive (Alix, TSG101, CD81) and EV-negative (GM130) markers. In the in vivo studies, PC-NVs successfully improved erectile function in cavernous nerve injury mice (∼82% of control values). Immunofluorescence staining showed significant increases in pericytes, endothelial cell, and neuronal contents. In the in vitro studies, PC-NVs significantly increased mouse cavernous endothelial cells tube formation, Schwann cell migration, and dorsal root ganglion and major pelvic ganglion neurite sprouting. Finally, Western blot analysis revealed that PC-NVs upregulated cell survival signaling (Akt and eNOS) and induced the expression of neurotrophic factors (brain-derived neurotrophic factor, neurotrophin-3, and nerve growth factor). Clinical Implications PC-NVs may be used as a strategy to treat erectile dysfunction after radical prostatectomy or in men with neurovascular diseases. Strengths & Limitations We evaluated the effect of PC-NVs in vitro and in a mouse nerve injury model, cavernous nerve injury. Additional studies are necessary to determine the detailed mechanisms of neurovascular improvement. Further study is needed to test whether PC-NVs are also effective when given weeks or months after nerve injury. Conclusion PC-NVs significantly improved erectile function by enhancing neurovascular regeneration. Local treatment with PC-NVs may represent a promising therapeutic strategy for the treatment of neurovascular diseases.

Pericyte‑derived extracellular vesicles‑mimetic nanovesicles improves peripheral nerve regeneration in mouse models of sciatic nerve transection

“…The diameter of the purified PC-NVs was ~30 nm. As previous studies have shown, the NVs also expressed EV surface markers, such as Alix, TSG101 and CD81 ( 17 , 18 ). Therefore, the extracted PC-NVs were characterized by western blot analysis.…”

“…Therefore, to overcome this limitation, a mini extruder system was developed, and >100-fold greater EV-mimetic NVs were extracted from different cell types, such as embryonic stem cells (ESCs) and mouse cavernous pericytes (MCPs). In addition, ESC-derived EV-mimetic NVs (ESC-NVs) and MCP-derived EV-mimetic NVs (PC-NVs) showed similar qualities to natural EVs, thus might be helpful for studies on neurovascular regeneration ( 17 , 18 ). However, the detailed molecular mechanisms by which PC-NVs promote nerve regeneration remains largely unknown.…”

“…Regeneration of severed nerves requires polarized vasculature induced by macrophages in the hypoxic bridge to guide the collective migration of Schwann cells ( 19 ). Our recent studies showed that ESC-NVs could rescue erectile function in diabetic mice, whereas PC-NVs significantly improved erectile dysfunction in a mouse cavernous nerve injury (CNI) model ( 17 , 18 ). However, there are no studies assessing the functional evaluation of PC-NVs in the peripheral nervous system (PNS), to the best of our knowledge.…”

RNA-sequencing profiling analysis of pericyte-derived extracellular vesicle–mimetic nanovesicles-regulated genes in primary cultured fibroblasts from normal and Peyronie’s disease penile tunica albuginea

“…These findings suggest that different antiangiogenic and molecular targets produce different results in the treatment of fibrosis. We recently reported in a mouse model of diabetic ED that pericyte-derived angiogenic factor restored erectile function by enhancing cavernous angiogenesis [ 14 , 15 ].…”

“…The cell-derived EV–mimetic NVs showed similar characteristics to the natural EVs [ 22 ]. We recently demonstrated that intracavernous injection of PC–NVs induced penile angiogenesis and rescued erectile function in a mouse model of cavernous nerve injury [ 15 ]. Along with potential relationship between angiogenesis and suppression of tissue fibrosis [ 11 – 13 ], the angiogenic potential of pericytes and PC–NVs led us to investigate the potential role of PC–NVs in the fibrogenic process of PD.…”