Exosomes from umbilical cord mesenchymal stromal cells promote the collagen production of fibroblasts from pelvic organ prolapse

BACKGROUND Pelvic organ prolapse(POP)involves pelvic organ herniation into the vagina due to pelvic floor tissue laxity,and vaginal structure is an essential factor.In POP,the vaginal walls exhibit abnormal collagen distribution and decreased fibroblast levels and functions.The intricate etiology of POP and the prohibition of trans-vaginal meshes in pelvic reconstruction surgery present challenges in targeted therapy development.Human umbilical cord mesenchymal stromal cells(hucMSCs)present limitations,but their exosomes(hucMSC-Exo)are promising therapeutic tools for promoting fibroblast proliferation and extracellular matrix remodeling.suppressed inflammation in POP group fibroblasts,stimulated primary fibroblast growth,and elevated collagen I(Col1)production in vitro.High-throughput RNA-seq of fibroblasts treated with hucMSC-Exo and miRNA sequencing of hucMSC-Exo revealed that abundant exosomal miRNAs downregulated matrix metalloproteinase 11(MMP11)expression.CONCLUSION HucMSC-Exo normalized the growth and function of primary fibroblasts from patients with POP by promoting cell growth and Col1 expression in vitro.Abundant miRNAs in hucMSC-Exo targeted and downregulated MMP11 expression.HucMSC-Exo-based therapy may be ideal for safely and effectively treating POP.

Experimental Effect of Umbilical Cord Mesenchymal Stem Cells on Stress Urinary Incontinence Rats Model and Efficient Tracking in vivo with MRI

Objective To assess the effect of intra-sphincteric injections of human umbilical cord mesenchymal stem cells(HUMSCs)on leak point pressure(LPP)changes in an animal model of stress urinary incontinence(SUI).Meanwhile,to investigate in vivo MRI tracking HUMSCs in SUI rats using a clinically available paramagnetic contrast agent(Gd-DTPA)and commercially available effentence transfection reagents..Materials and Methods HUMSCs were dual labeled with Gd-DTPA and PKH26,the labeling efficiency and longevity of Gd-DTPA maintenance were measured and cell viability and proliferation were assessed.39 female Sprague–Dawley SUI rats.12 normal rats and 12 SUI rats received periurethral injection of PBS and 12 SUI rats were given periurethral injection of dual labeled HUMSCs.3 SUI rat sreceived periurethral injection of u nlabeled HUMSCs.Six weeks after injection,LPP was undertaken in animals.All rats were sacrificed and frozen urethra sections were submitted to pathology and immunohistochemistry assessment.Results The labeling efficiency of Gd-DTPA was up to 80%,the labeling procedure did not influence cell viability and proliferation.The signal intensity on T1-weighted imaging and T1 values of labeled cells were significantly higher than those of unlabeled cells.In vitro,differentiated HUMSCs expressed myosin heavy chain(MHC)and desmin,markers of striated muscles.In vivo,immunohistochemistry of rat urethras revealed dual labeled HUMSCs in situ and at the injection site.LPP was significantly improved in animals injected with HUMSCs.Atrophic urethras with implanted HUMSCs were positively stained for MHC and desmin.The distribution and migration of labeled cells could be tracked by MRI more than 14 days after t ransplantation.Conclusion HUMSCs have the ability to differentiate striated muscles,as demonstrated by MHC and desmin expression.Periurethral injection of HUMSCs in an animal model of SUI restored the damaged external urethral sphincter and significantly improved LPP.MRI can track Gd-DTPA–labled HUMSCs in an animal model of SUI in vivo.

Amelioration of ligamentum flavum hypertrophy using umbilical cord mesenchymal stromal cell-derived extracellular vesicles

Ligamentum flavum(LF)hypertrophy(LFH)has been recognised as one of the key contributors to lumbar spinal stenosis.Currently,no effective methods are available to ameliorate this hypertrophy.In this study,human umbilical cord mesenchymal stromal cell-derived extracellular vesicles(hUCMSC-EVs)were introduced for the first time as promising vehicles for drug delivery to treat LFH.The downregulation of miR-146a-5p and miR-221-3p expressions in human LF tissues negatively correlated with increased LF thickness.The hUCMSC-EVs enriched with these two miRNAs significantly suppressed LFH in vivo and notably ameliorated the progression of transforming growth factorβ1(TGF-β1)-induced fibrosis in vitro after delivering these two miRNAs to mouse LF cells.The results further demonstrated that miR-146a-5p and miR-221-3p directly bonded to the 3′-UTR regions of SMAD4 mRNA,thereby inhibiting the TGF-β/SMAD4 signalling pathway.Therefore,this translational study determined the effectiveness of a hUCMSC-EVs-based approach for the treatment of LFH and revealed the critical target of miR-146a-5p and miR-221-3p.Our findings provide new insights into promising therapeutics using a hUCMSC-EVs-based delivery system for patients with lumbar spinal stenosis.

Multipotent mesenchymal stromal cells are fully permissive for human cytomegalovirus infection

Congenital human cytomegalovirus(HCMV) infection is a leading infectious cause of birth defects.Previous studies have reported birth defects with multiple organ maldevelopment in congenital HCMV-infected neonates. Multipotent mesenchymal stromal cells(MSCs) are a group of stem/progenitor cells that are multi-potent and can self-renew, and they play a vital role in multiorgan formation. Whether MSCs are susceptible to HCMV infection is unclear. In this study, MSCs were isolated from Wharton's jelly of the human umbilical cord and identified by their plastic adherence, surface marker pattern, and differentiation capacity. Then, the MSCs were infected with the HCMV Towne strain, and infection status was assessed via determination of viral entry,replication initiation, viral protein expression, and infectious virion release using western blotting,immunofluorescence assays, and plaque forming assays. The results indicate that the isolated MSCs were fully permissive for HCMV infection and provide a preliminary basis for understanding the pathogenesis of HCMV infection in non-nervous system diseases, including multi-organ malformation during fetal development.