CCR2-Overexpressing Mesenchymal Stem Cells Targeting Damaged Liver Enhances Recovery of Acute Liver Failure

Abstract Background: Mesenchymal stem cell (MSC) transplantation is emerging as a promising cell therapeutic strategy in acute liver failure (ALF) clinical research. The potency of MSCs to migrate and engraft into targeted lesions could largely determine their clinical efficacy, in which chemokine/receptor axes play a crucial role. Unfortunately, the downregulation of chemokine receptors expression after in vitro expansion results in a poor homing capacity of MSCs.Methods: By evaluating the chemokine expression profile in the liver of ALF patients and ALF mice, we found that CCL2 expression was highly upregulated in damaged livers, while the corresponding receptor, CCR2, was lacking in cultured MSCs. Thus, we genetically modified MSCs to overexpress CCR2 and investigated the targeted homing capacity and treatment efficacy of MSCCCR2 compared to those of the MSCvector control.Results: In vivo and ex vivo near-infrared fluorescence imaging showed that MSCCCR2 rapidly migrated and localized to injured livers in remarkably greater numbers following systemic infusion, and these cells were retained in liver lesions for a longer time than MSCvector. Furthermore, MSCCCR2 exhibited significantly enhanced efficacy in the treatment of ALF in mice, which was indicated by a dramatically improved survival rate, the alleviation of liver injury with reduced inflammatory infiltration, and hepatic apoptosis, and the promotion of liver regeneration.Conclusions: Altogether, these results indicate that CCR2 overexpression enhances the targeted migration of MSCs to damaged livers, improves their treatment effect, and may provide a novel strategy for improving the efficacy of cell therapy for ALF.

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Efficient Photoacoustic Imaging With Biomimetic Mesoporous Silica-Based Nanoparticles

Frontiers in Bioengineering and Biotechnology ◽

10.3389/fbioe.2021.762956 ◽

2021 ◽

Vol 9 ◽

Author(s):

Chuangjia Huang ◽

Xiaoling Guan ◽

Hui Lin ◽

Lu Liang ◽

Yingling Miao ◽

...

Keyword(s):

Mesoporous Silica ◽

Near Infrared ◽

Ex Vivo ◽

Photoacoustic Imaging ◽

Mesoporous Silica Nanoparticles ◽

Good Biocompatibility ◽

Targeting Ability ◽

Body Clearance

Indocyanine green (ICG), a near-infrared (NIR) fluorescent dye approved by the Food and Drug Administration (FDA), has been extensively used as a photoacoustic (PA) probe for PA imaging. However, its practical application is limited by poor photostability in water, rapid body clearance, and non-specificity. Herein, we fabricated a novel biomimetic nanoprobe by coating ICG-loaded mesoporous silica nanoparticles with the cancer cell membrane (namely, CMI) for PA imaging. This probe exhibited good dispersion, large loading efficiency, good biocompatibility, and homologous targeting ability to Hela cells in vitro. Furthermore, the in vivo and ex vivo PA imaging on Hela tumor-bearing nude mice demonstrated that CMI could accumulate in tumor tissue and display a superior PA imaging efficacy compared with free ICG. All these results demonstrated that CMI might be a promising contrast agent for PA imaging of cervical carcinoma.

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Pinocembrin ameliorates acute liver failure via activating the Sirt1/PPARα pathway in vitro and in vivo

European Journal of Pharmacology ◽

10.1016/j.ejphar.2021.174610 ◽

2021 ◽

pp. 174610

Author(s):

Pan Cao ◽

Qian Chen ◽

Chunxia Shi ◽

Maohua Pei ◽

Luwen Wang ◽

...

Keyword(s):

Liver Failure ◽

Acute Liver Failure

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IN VIVO NEAR-INFRARED FLUORESCENCE IMAGING OF HUMAN COLON ADENOCARCINOMA BY SPECIFIC IMMUNOTARGETING OF A TUMOR-ASSOCIATED MUCIN

Journal of Innovative Optical Health Sciences ◽

10.1142/s1793545809000759 ◽

2009 ◽

Vol 02 (04) ◽

pp. 407-422 ◽

Cited By ~ 3

Author(s):

RALPH S. DACOSTA ◽

YING TANG ◽

TUULA KALLIOMAKI ◽

RAYMOND M. REILLY ◽

ROBERT WEERSINK ◽

...

Keyword(s):

Fluorescence Microscopy ◽

Fluorescence Imaging ◽

Normal Tissue ◽

Near Infrared ◽

Ex Vivo ◽

Colon Adenocarcinoma ◽

Human Colon ◽

Human Colon Adenocarcinoma

Background and Aims: Accurate endoscopic detection of premalignant lesions and early cancers in the colon is essential for cure, since prognosis is closely related to lesion size and stage. Although it has great clinical potential, autofluorescence endoscopy has limited tumor-to-normal tissue image contrast for detecting small preneoplastic lesions. We have developed a molecularly specific, near-infrared fluorescent monoclonal antibody (CC49) bioconjugate which targets tumor-associated glycoprotein 72 (TAG72), as a contrast agent to improve fluorescence-based endoscopy of colon cancer. Methods: The fluorescent anti-TAG72 conjugate was evaluated in vitro and in vivo in athymic nude mice bearing human colon adenocarcinoma (LS174T) subcutaneous tumors. Autofluorescence, a fluorescent but irrelevant antibody and the free fluorescent dye served as controls. Fluorescent agents were injected intravenously, and in vivo whole body fluorescence imaging was performed at various time points to determine pharmacokinetics, followed by ex vivo tissue analysis by confocal fluorescence microscopy and histology. Results: Fluorescence microscopy and histology confirmed specific LS174T cell membrane targeting of labeled CC49 in vitro and ex vivo. In vivo fluorescence imaging demonstrated significant tumor-to-normal tissue contrast enhancement with labeled-CC49 at three hours post injection, with maximum contrast after 48 h. Accumulation of tumor fluorescence demonstrated that modification of CC49 antibodies did not alter their specific tumor-localizing properties, and was antibody-dependent since controls did not produce detectable tumor fluorescence. Conclusions: These results show proof-of-principle that our near-infrared fluorescent-antibody probe targeting a tumor-associated mucin detects colonic tumors at the molecular level in real time, and offer a basis for future improvement of image contrast during clinical fluorescence endoscopy.

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Nanocurcumin-Loaded UCNPs for Cancer Theranostics: Physicochemical Properties, In Vitro Toxicity, and In Vivo Imaging Studies

Nanomaterials ◽

10.3390/nano11092234 ◽

2021 ◽

Vol 11 (9) ◽

pp. 2234

Author(s):

Anbharasi Lakshmanan ◽

Roman A. Akasov ◽

Natalya V. Sholina ◽

Polina A. Demina ◽

Alla N. Generalova ◽

...

Keyword(s):

Near Infrared ◽

Therapeutic Potential ◽

Ex Vivo ◽

Laboratory Animals ◽

In Vitro Toxicity ◽

Upconversion Nanoparticles ◽

Lewis Lung Cancer ◽

Spectral Bands

Formulation of promising anticancer herbal drug curcumin as a nanoscale-sized curcumin (nanocurcumin) improved its delivery to cells and organisms both in vitro and in vivo. We report on coupling nanocurcumin with upconversion nanoparticles (UCNPs) using Poly (lactic-co-glycolic Acid) (PLGA) to endow visualisation in the near-infrared transparency window. Nanocurcumin was prepared by solvent-antisolvent method. NaYF4:Yb,Er (UCNP1) and NaYF4:Yb,Tm (UCNP2) nanoparticles were synthesised by reverse microemulsion method and then functionalized it with PLGA to form UCNP-PLGA nanocarrier followed up by loading with the solvent-antisolvent process synthesized herbal nanocurcumin. The UCNP samples were extensively characterised with XRD, Raman, FTIR, DSC, TGA, UV-VIS-NIR spectrophotometer, Upconversion spectrofluorometer, HRSEM, EDAX and Zeta Potential analyses. UCNP1-PLGA-nanocurcumin exhibited emission at 520, 540, 660 nm and UCNP2-PLGA-nanocurmin showed emission at 480 and 800 nm spectral bands. UCNP-PLGA-nanocurcumin incubated with rat glioblastoma cells demonstrated moderate cytotoxicity, 60–80% cell viability at 0.12–0.02 mg/mL marginally suitable for therapeutic applications. The cytotoxicity of UCNPs evaluated in tumour spheroids models confirmed UCNP-PLGA-nanocurcumin therapeutic potential. As-synthesised curcumin-loaded nanocomplexes were administered in tumour-bearing laboratory animals (Lewis lung cancer model) and showed adequate contrast to enable in vivo and ex vivo study of UCNP-PLGA-nanocurcumin bio distribution in organs, with dominant distribution in the liver and lungs. Our studies demonstrate promise of nanocurcumin-loaded upconversion nanoparticles for theranostics applications.

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Abstract 17216: Intracoronary Dual-modal OCT/NIRF Structural-Molecular Imaging with a Clinical Dose of Indocyanine Green (ICG) for Detection of High-risk Coronary Plaques in Diabetic Swine Model

Circulation ◽

10.1161/circ.130.suppl_2.17216 ◽

2014 ◽

Vol 130 (suppl_2) ◽

Author(s):

Sunwon Kim ◽

Min Woo Lee ◽

Han Saem Cho ◽

Joon Woo Song ◽

Sunki Lee ◽

...

Keyword(s):

High Risk ◽

Near Infrared ◽

Ex Vivo ◽

Swine Model ◽

Fully Integrated ◽

Coronary Syndrome ◽

Fibrous Cap ◽

Nirf Imaging

Background: Acute coronary syndrome is frequently caused by rupture of macrophage abundant plaques with a large lipid-rich core. The present study aimed to investigate whether a fully integrated OCT/NIRF imaging combined with a clinically available near-infrared fluorescence (NIRF) enhancing ICG can detect the inflamed, lipid-rich plaques in swine coronary atheromata whose phenotype is similar to human vulnerable fibroatheroma. Methods and Results: Accelerated atherosclerosis was made by coronary balloon denudation in alloxan induced diabetic minipigs. A rapid coronary imaging (20 mm/sec pullback speed) using a fully integrated OCT/NIRF catheter was safely performed 30 minutes after I.V. injection of ICG (2.0 mg/kg) just under contrast purge. OCT clearly identified the lipid-rich plaques with fibrous cap. Simultaneously acquired, distance-calibrated NIRF imaging detected lipid-laden macrophage signals in OCT-proven plaques (figure). The in vivo plaque target-to-background ratio (pTBR) was significantly higher in ICG-injected swine compared to non-diabetic swines or saline-injected controls (p<0.05), which was validated on ex vivo fluorescence reflectance imaging (FRI) (figure). The in vivo and ex vivo peak pTBRs correlated significantly (p<0.05). In vitro experiments, and histopathology including fluorescence microscopic imaging and immunostaining of the plaque sections corroborated the findings in vivo . Conlusions: An OCT/NIRF imaging with a clinical use of ICG accurately identified macrophage abundant, lipid-rich coronary plaques in diabetic atheromatous minipigs. This highly translatable dual-modal molecular-structural imaging could be relevant for clinical intracoronary detection of high-risk plaques.

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FTY720 Abrogates the Therapeutic Potential of Adoptively Transferred Treg Via Inhibition of IL-2 Induced in Vivo Expansion

Blood ◽

10.1182/blood.v112.11.2584.2584 ◽

2008 ◽

Vol 112 (11) ◽

pp. 2584-2584

Author(s):

Anna Maria Wolf ◽

Kathrin Hochegger ◽

Robert Zeiser ◽

Christoph Duerr ◽

Michael Sixt ◽

...

Keyword(s):

T Cells ◽

Adoptive Transfer ◽

Chemokine Receptors ◽

Therapeutic Potential ◽

Ex Vivo ◽

Effective Means ◽

Effector T Cells ◽

Target Tissue

Abstract CD4+CD25+ T cells (Treg) entry into secondary lymphoid organs (SLO) and local expansion after activation is at least in part responsible for their immunosuppressive action. Thus we hypothesized that trapping of adoptively transferred Treg in SLO would be an effective means to tip the balance towards a more immunosuppressive milieu within the LN microenvironment. Systemic application of the sphingosine-phosphate receptor agonist FTY720 has been proven to trap harmful effector T cells in SLO, thereby inhibiting their migration and destruction of target tissue. Here we provide first evidence that selective entrapment of adoptively transferred Treg in inflammatory LN can be achieved by blockade of SP-receptors upon ex vivo exposure of Treg to FTY720 before adoptive transfer. FTY720 exposure did not interfere with proper Treg localization within the T-cell areas of SLO as determined by immunofluorescent microscopy after co-transfer of either FTY720- or solvent exposed and subsequently differentially labelled Treg. However, despite the fact that the in vitro phenotype (including expression of adhesion and chemokine receptors), function (including anergy and suppressive activity) and survival (determined by Annexin/PI staining) of Treg remained unaltered by FTY720, it abrogated their protective effect after adoptive transfer in a murine model of acute experimental glomerulonephritis (determined by quantification of proteinuria and histological analysis) as well as in an acute GvHD model (determined by survival analysis and quantification of the in vivo expansion of luciferase-transgenic effector T cells by bioluminiscence technology). Notably, adoptive transfer of CFSE-labelled Treg revealed a markedly impaired proliferation of Treg in inflammatory SLO when pre-exposed to FTY720 ex vivo. Accordingly, FTY720 blocked Treg-proliferation induced by TCR-stimulation in combination with IL-2 in vitro. In line with this observation, FTY720 completely abolishes IL-2 induced phosphorylation of STAT-5. Thus, SP-1P receptors induce Treg trapping in inflammatory SLO but abrogate their in vivo immunosuppressive potential by inhibition of local Treg expansion.

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The Effects of Conditioned Medium Derived from Mesenchymal Stem Cells Cocultured with Hepatocytes on Damaged Hepatocytes and Acute Liver Failure in Rats

Stem Cells International ◽

10.1155/2018/9156560 ◽

2018 ◽

Vol 2018 ◽

pp. 1-10 ◽

Cited By ~ 3

Author(s):

Li Chen ◽

Jiexin Zhang ◽

Lu Yang ◽

Guoying Zhang ◽

Yingjie Wang ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Liver Failure ◽

Acute Liver Failure ◽

Conditioned Medium ◽

Therapeutic Potential ◽

L02 Cells ◽

Cells Cultured

Mesenchymal stem cells (MSCs) and hepatocytes are two attractive sources of cell-based therapies for acute liver failure (ALF). The cotransplantation of hepatocytes with MSCs can improve the therapeutic performance for the treatment of ALF. However, the therapeutic potential of conditioned medium (CM) derived from MSCs cocultured with hepatocytes (MSC-H-CM) remains unclear. The purpose of this study was to investigate the effects of MSC-H-CM on damaged hepatocytes in vitro and on D-galactosamine-induced ALF in vivo. D-Galactosamine-treated L02 cells cultured in MSC-H-CM exhibited higher of cell viability and total protein synthesis than L02 cells cultured in MSC-CM, CM derived from hepatocytes (H-CM), MSC-CM + H-CM, or with nonconditioned medium (NCM). Lactate dehydrogenase and aspartate aminotransferase levels were lower in the supernatant of damaged L02 cells cultured in MSC-H-CM than in that of L02 cells cultured in other types of CM. The lowest percentage of apoptotic cells was observed after the MSC-H-CM treatment. When CM was injected into the tail vein of rats with ALF, MSC-H-CM was the most successful at preventing the release of liver injury biomarkers and in promoting the recovery of liver structure. The greatest survival rate 7 days after the first treatment was observed in the MSC-H-CM-treated rats. Our results reveal that the delivery of MSC-H-CM could be a novel strategy for integrating the therapeutic potentials of hepatocytes and MSCs for the treatment of ALF.

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Liposomal Chitosan-N-Acetyl Cysteine Nanoparticle for In-Vitro/In-Vivo Near Infrared Fluorescent (NIR) Imaging

10.26434/chemrxiv.9976145.v1 ◽

2019 ◽

Author(s):

Piyush Kumar ◽

Timothy Van Treuren ◽

Amalendu Ranjan ◽

Jamboor K Vishwanatha

Keyword(s):

Near Infrared ◽

Infrared Imaging ◽

Tumor Imaging ◽

Ex Vivo ◽

Breast Cancer Cell Lines ◽

Nir Imaging ◽

Negative Surface ◽

Acetyl Cysteine

<p>In this work, we studied the effect of chitosan conjugated N-acetyl cysteine (CHT-NAC) coating on liposomal (DMPC:14 lysoPG: DSPE-2000-NH2) nanoparticles as a vehicle to cross the blood-brain barrier. The size of lipo-NP and Lipo-CHT-NAC NP were sub 50 nm with negative surface charge consistent with its use in an intravenous application. In vitro near infrared (NIR) imaging showed good cellular uptake in two triple-negative breast cancer cell lines (MDA-MB-231 and brain metastatic MDA-MB-831). Live (4-120 h) and ex-vivo near-infrared imaging at 24 h in nude mice showed the extended circulation of CHT-NAC Lipo-NP. These results demonstrated that Lipo-CHT-NAC NP could be used for metastatic brain tumor imaging.</p>

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Liposomal Chitosan-N-Acetyl Cysteine Nanoparticle for In-Vitro/In-Vivo Near Infrared Fluorescent (NIR) Imaging

10.26434/chemrxiv.9976145 ◽

2019 ◽

Author(s):

Piyush Kumar ◽

Timothy Van Treuren ◽

Amalendu Ranjan ◽

Jamboor K Vishwanatha

Keyword(s):

Near Infrared ◽

Infrared Imaging ◽

Tumor Imaging ◽

Ex Vivo ◽

Breast Cancer Cell Lines ◽

Nir Imaging ◽

Negative Surface ◽

Acetyl Cysteine

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Fusobacterium Nucleatum Promotes the Development of Acute Liver Failure by Inhibiting the NAD+ Salvage Metabolic Pathway

10.21203/rs.3.rs-1091141/v1 ◽

2021 ◽

Author(s):

Pan Cao ◽

Qian Chen ◽

Chunxia Shi ◽

Luwen Wang ◽

Zuojiong Gong

Keyword(s):

Energy Metabolism ◽

Liver Failure ◽

Acute Liver Failure ◽

Fusobacterium Nucleatum ◽

Mucosal Barrier ◽

Liver Inflammation ◽

Ampk Signaling ◽

Inflammatory Damage

Abstract Background: Acute liver failure (ALF) patients are often accompanied by severe energy metabolism abnormalities and intestinal microecological imbalance. The intestinal mucosal barrier is severely damaged. Intestinal endotoxin can induce intestinal endotoxemia through the "Gut-Liver axis". More and more evidence shows that members of the gut microbiota, especially Fusobacterium nucleatum (F. nucleatum), are related to inflammatory bowel disease, but whether F. nucleatum is involved in the development of ALF and whether it affects the liver energy metabolism is unclear. Methods: This study first detected the abundance of F. nucleatum and its effect on ALF disease, and explored whether F. nucleatum aggravated liver inflammation in vitro and in vivo. Results: Our data showed that liver tissues of ALF patients contained different abundances of F. nucleatum, which were related to the degree of liver inflammation. In addition, we found that F. nucleatum infection affected the energy metabolism of the liver during the development of ALF, inhibited the synthesis pathway of nicotinamide adenine dinucleotide (NAD+)'s salvage metabolism, and promoted inflammatory damage in the liver. In terms of mechanism, F. nucleatum inhibited NAD+ and the NAD+-dependent SIRT1/AMPK signaling pathway, and promoted liver damage of ALF. Conclusions: F. nucleatum coordinates a molecular network including NAD+ and SIRT1 to control the progress of ALF. Detection and targeting of F. nucleatum and its related pathways may provide valuable insights for the treatment of ALF.

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