This study aims to evaluate the effectiveness and safety of the spheroid reservoir bioartificial liver (SRBAL) with porcine hepatocyte organoids in a preclinical nonhuman primate model of acute liver failure (ALF).Methods: Thirty healthy rhesus monkeys were infused with α-amanitin and lipopolysaccharide and randomized into five groups (ALF alone control group; sham no-cell SRBAL treatment group; groups A, B and C with SRBAL treatment started at 12 h, 24 h and 36 h after induction of ALF, respectively). Animals were continuously treated with the SRBAL device for 6 h and followed for up to 336 h.Results: Survival of ALF monkeys improved with hepatocyte SRBAL treatment compared to control groups. Blood ammonia and total bilirubin were lower, and albumin levels were higher in all hepatocyte SRBAL treatment groups. No evidence of porcine endogenous retrovirus was identified in monkey liver or blood after SRBAL treatment. Titers of monkey antibody (IgG, IgM) did not rise after SRBAL treatment. In survival cases, the proportion of necrotic and apoptotic hepatocytes was lower in SRBAL-treated groups, with earlier liver regeneration leading to recovery. Cytokines TNF-α, IL-6, IL-12, IL-1β, IL-8, IFN-γ and IL-2 were ameliorated by the SRBAL treatment, while levels of M-CSF; HGF, EGF and VEGF; IL-1RA and MIF rose on priming, proliferation and the late phase of liver regeneration.Conclusions: The benefit of SRBAL therapy included preventive effects and therapeutic effects. SRBAL improved survival rate and prolonged median survival time in a nonhuman primate model of drug-induced ALF, and these benefits declined with a delay in the initiation of therapy. Improved survival and recovery of ALF monkeys was associated with a reduction in blood ammonia levels, inhibition of the pro-inflammatory response of ALF, and provided a microenvironment more suitable for regeneration of the injured liver.

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Background The challenge of using bioengineered liver lies in sustaining the quantity of high‐quality hepatocytes and the vasculature for blood perfusion. We characterized the heparinization of a porcine decellularized liver scaffold (DLS) as a carrier to support hepatocyte angiogenesis, thereby developing functional and vascularized hepatic tissue useful to treat liver injury. Method The porcine DLS was obtained by the removal of cellular components and then subjected to heparinization by the end‐point attachment technique. The heparinized DLSs were recellularized with rat hepatocyte spheroids to construct engineered hepatic tissue. The hepatic tissue was heterotopically implanted in the omentum majus of a rat model with liver injury induced by carbon tetrachloride (CCl4). Results Hepatocyte spheroids in the heparinized DLS remained viable for at least 10 weeks in vivo. The entire scaffold was populated with hepatocytes and arranged well. The volume of the heparinized DLS group was expanded over 400‐fold. Liver‐specific functions such as albumin synthesis, glycogen storage and cytochrome P 3A4 activity were highly expressed in the hepatic tissue. In addition, endothelial cells were recruited, as shown by CD31 staining, and new blood vessels formed, as visualized by fluorescein isothiocyanate‐labelled dextran intravital confocal microscopy. The heparinized bioengineered hepatic tissue alleviated CCl4‐induced liver injury by regulating the deposition and degradation of the extracellular matrix. Conclusion Primary hepatocyte spheroids survived for an extended time on the heparinized DLS and expanded to generate vascularized and functional bioengineered hepatic tissue that can alleviate liver injury in rats.

BackgroundDespite an increasing understanding of chronic obstructive pulmonary disease (COPD) pathogenesis, the mechanisms of diverse cell populations in the human lung remain unknown. Using single-cell RNA sequencing (scRNA-Seq), we can reveal changes within individual cell populations in COPD that are important for disease pathogenesis and characteristics. MethodsWe performed scRNA-Seq on lung tissue obtained from donors with non-COPD and mild-to-moderate COPD to identify disease-related genes within different cell types. We testified the findings using qRT−PCR, immunohistochemistry, immunofluorescence and Western blotting from 25 additional subjects and RAW 264.7 macrophages. Targeting ferroptosis with the ferroptosis inhibitor ferrostatin-1, iron chelator deferoxamine or HO-1 inhibitor zinc protoporphyrin was administered in the experimental cigarette smoke COPD mouse model.ResultsWe identified two populations of alveolar macrophages (AMs) in the human lung that were dysregulated in COPD patients. We discovered that M2-like AMs modulate susceptibility to ferroptosis by disrupting lipid and iron homeostasis both in vivo and in vitro. The discrepancy in sensitivity to ferroptosis can be determined and regulated by HO-1. In contrast, M1-like AMs showed the ability to attenuate oxidative stress and exert resistance to ferroptosis. In addition, the expression of genes within M2-like AMs is also involved in defects in phagocytosis and lysosome distortion. This ferroptotic phenotype was ameliorated by antiferroptotic compounds, iron chelators and HO-1 inhibitors. During COPD, the accumulation of lipid peroxidation drives ferroptosis-sensitive M2-like AMs, while M1-like AMs show characteristics of ferroptosis resistance. Ferroptotic M2 AMs lose their anti-inflammatory and repair functions but provoke inflammatory responses, resulting in consistent inflammation and tissue damage in the presence of M1 AMs in COPD. ConclusionAppropriate interventions in ferroptosis can reduce the occurrence of infections and acute onset, and delay the COPD process.

WHAT THIS PAPER ADDSThis cohort study with inverse probability of treatment weighting analyses suggests that endovascular aortic aneurysm repair (EVAR) with crossed limb (CL) configuration is safe and effective in general patients with abdominal aortic aneurysm and may decrease the risk of type IB endoleak in patients with a large aneurysm sac or tortuous iliac arteries. However, the application of the CL technique in patients with a severely angulated neck may be associated with a higher risk of re-intervention due to potential higher risks of adverse limb events.Objective: The aim was to compare mid term outcomes between crossed limb (CL) and standard limb (SL) configuration in patients who underwent endovascular aortic aneurysm repair (EVAR). Methods: This was a comparative cohort study. Eligible patients who underwent EVAR between September 2011 and March 2019 in a tertiary academic centre were included. Inverse probability of treatment weighting (IPTW) was used to balance the demographic, anatomical and operative baseline characteristics between the two groups. The primary outcome was adverse limb events including type IB endoleak (T1BEL), type III endoleak, and limb occlusion. Cox proportional hazards regression and marginal structural model were performed to compare time to event outcomes. Results: The study included 729 patients (194 CL and 535 SL) with a median follow up of 34 months (interquartile range 16 e 62 months). The weighted analyses revealed no significant difference between CL and SL EVAR in terms of adverse limb events, type IA endoleak (T1AEL), type II endoleak (T2EL), re-intervention, and overall survival. In the subgroup analysis of large aneurysm sac, the CL configuration was associated with a significantly decreased risk of T1BEL (hazard ratio [HR] 0.31, 95% confidence interval [CI] 0.12 e 0.78, p ¼ .014). Similar results were also observed in the subgroup of tortuous iliac arteries (HR 0.30, 95% CI 0.11 e 0.81, p ¼ .017). After stratification by severe neck angulation, no significant difference was found between CL and SL EVAR for T1AEL, but the CL configuration was associated with a significantly increased risk of reintervention (HR 2.69, 95% CI 1.31 e 5.51, p ¼ .007). In addition, a trend towards a higher risk of adverse limb events in the CL group with severely angulated proximal neck was observed. Conclusion: CL configuration in EVAR is safe and may be associated with a lower risk of T1BEL in patients with a large aneurysm sac or tortuous iliac arteries. However, it should be applied cautiously to aneurysms with a severely angulated neck due to the potentially higher risk of re-intervention.