| Objective: To observe the therapeutic effect of DF-PEG/GCS selfhealing hydrogel seeded with adipose-derived stem cells(ASCs)and EVs on erectile dysfunction(ED)in diabetic rats,and investigate the underlying mechanisms.Study content: DF-PEG/GCS hydrogels were prepared and evaluated.Next,ASCs were seeded into the DF-PEG/GCS hydrogels.Morphological,histological,functional and tissue regeneration of corpus cavernousa after penile cavernous injection treatment was evaluated in a rat model of Diabetes mellitus erectile dysfunction(DMED).The specific mechanisms of vascularization and smooth muscle regeneration were investigated.Methods: Adipose stem cells were extracted from 2 week old SpragueDawley rats.Intracavernous injection was carried out with a hydrogel.After 12 weeks,the intracavernous pressure(ICP)and mean arterial pressure(MAP)were measured.The cavernosal smooth muscle regeneration was determined by Masson staining.The number of blood vessels was detected by immunofluorescent staining.The controlled-release effects of hydrogels were evaluated by whole body imaging system.Extracellular vesicles were obtained from adipose-derived stem cells by ultracentrifugation.Then EVs were seeded into hydrogels and injected intracavernally.The ICP/MAP,immunofluorescence and Masson staining were again evaluated to observe the recovery of erectile function and vascularization.And smooth muscle regeneration was evaluated by immunofluorescent staining.Results: ASCs can partially restore the erectile function of diabetic ED rats,while DF-PEG/GCS hydrogel can further improve erectile function.Meanwhile,whole-body in vivo imaging confirms the control-released function of hydrogels on ASCs.Western Blot showed that ASCs can ameliorate the inflammation of the corpus cavernosum and promote vascularization and neuronization.PCR showed that ASCs prevent cells from apoptosis,and immunofluorescence showed that ASCs promote vascularization and smooth muscle regeneration.DF-PEG/GCS hydrogels enable ASCs to exert their effects more efficiently.EVs can improve the erectile function of diabetic ED rats,while DF-PEG/GCS hydrogels can further promote the effects of EVs.Masson staining and immunofluorescence showed that EVs can promote vascularization and neuronalization of the corpus cavernosum and also promote smooth muscle cell regeneration,which proved to be a good therapeutic substitute for ASCs.Conclusion: DF-PEG/GCS self-healing hydrogels seeded with adipose-derived stem cells and extracellular vesicles can restore erectile function in diabetic ED rats.Hydrogels can help ASCs to be sustainedly released in corpus cavernosum,which promote vascularization,neurogenesis and smooth muscle regeneration.Hydrogels seed with EVs can further improve erectile function in diabetic ED rats. |
PDF Full Text Request