Portal Fibrotic Cord is Associated with Transjugular Intrahepatic Portosystemic Shunt Failure and Death in Cirrhotic Patients

Background and Aims:Occlusive portal vein thrombosis(PVT)often causes portal hypertension-related complications in cirrhotic patients.Transjugular intrahepatic portosystemic shunt(TIPS)is an effective treatment for this difficult problem.However,the factors influencing TIPS success and overall survival in patients with occlusive PVT are unknown.This study investigated the factors influencing TIPS success and overall survival in cirrhotic patients with occlusive PVT.Methods:Cirrhotic patients with occlusive PVT were selected from a prospective database of consecutive patients treated with TIPS in Xijing Hospital between January 2015 and May 2021.Baseline characteristics,TIPS success rate,complications,and survival were collected,and the factors associated with the TIPS success rate and transplant-free survival were analyzed.Results:A total of 155 cirrhotic patients with occlusive PVT were enrolled.TIPS succeeded in 126(81.29%)cases.The 1-year survival rate was 74%.Compared with those without,patients with portal fibrotic cord had a lower TIPS success rate(39.02%vs.96.49%,p<0.001),shorter median overall survival(300 vs.1,730 days,p<0.001)and more operation-related complications(12.20%vs.1.75%,p<0.01).Logistic regression analysis found that portal fibrotic cord(odds ratio 0.024)was a risk factor for TIPS failure.Univariate and multivariate analysis showed that portal fibrotic cord was an independent predictor of death(hazard ratio 2.111;95%CI:1.094-4.071,p=0.026).Conclusions:Portal fibrotic cord increased the TIPS failure rate and is a risk factor for poor prognosis in cirrhotic patients.

Extracellular vesicles from 3D cultured dermal papilla cells improve wound healing via Krüppel-like factor 4/vascular endothelial growth factor A-driven angiogenesis

Background:Non-healing wounds are an intractable problem of major clinical relevance.Evidence has shown that dermal papilla cells(DPCs)may regulate the wound-healing process by secreting extracellular vesicles(EVs).However,low isolation efficiency and restricted cell viability hinder the applications of DPC-EVs in wound healing.In this study,we aimed to develop novel 3D-DPC spheroids(tdDPCs)based on self-feeder 3D culture and to evaluate the roles of tdDPC-EVs in stimulating angiogenesis and skin wound healing.Methods:To address the current limitations of DPC-EVs,we previously developed a self-feeder 3D culture method to construct tdDPCs.DPCs and tdDPCs were identified using immunofluorescence staining and flow cytometry.Subsequently,we extracted EVs from the cells and compared the effects of DPC-EVs and tdDPC-EVs on human umbilical vein endothelial cells(HUVECs)in vitro using immunofluorescence staining,a scratch-wound assay and a Transwell assay.We simultaneously established a murine model of full-thickness skin injury and evaluated the effects of DPC-EVs and tdDPC-EVs on wound-healing efficiency in vivo using laser Doppler,as well as hematoxylin and eosin,Masson,CD31 andα-SMA staining.To elucidate the underlying mechanism,we conducted RNA sequencing(RNA-seq)of tdDPC-EV-and phosphate-buffered saline-treated HUVECs.To validate the RNA-seq data,we constructed knockdown and overexpression vectors of Krüppel-like factor 4(KLF4).Western blotting,a scratch-wound assay,a Transwell assay and a tubule-formation test were performed to detect the protein expression,cell migration and lumen-formation ability of KLF4 and vascular endothelial growth factor A(VEGFA)in HUVECs incubated with tdDPC-EVs after KLF4 knockdown or overexpression.Dual-luciferase reporter gene assays were conducted to verify the activation effect of KLF4 on VEGFA.Results:We successfully cultured tdDPCs and extracted EVs from DPCs and tdDPCs.The tdDPC-EVs significantly promoted the proliferation,lumen formation and migration of HUVECs.Unlike DPC-EVs,tdDPC-EVs exhibited significant advantages in terms of promoting angiogenesis,accelerating wound healing and enhancing wound-healing efficiency both in vitro and in vivo.Bioinformatics analysis and further functional experiments verified that the tdDPC-EV-regulated KLF4/VEGFA axis is pivotal in accelerating wound healing.Conclusions:3D cultivation can be utilized as an innovative optimization strategy to effectively develop DPC-derived EVs for the treatment of skin wounds.tdDPC-EVs significantly enhance wound healing via KLF4/VEGFA-driven angiogenesis.