Proinflammatory Effect of Carbon-Based Nanomaterials: In Vitro Study on Stimulation of Inflammasome NLRP3 via Destabilisation of Lysosomes

Carbon-based nanomaterials (C-BNM) have recently attracted an increased attention as the materials with potential applications in industry and medicine. Bioresistance and proinflammatory potential of C-BNM is the main obstacle for their medicinal application which was documented in vivo and in vitro. However, there are still limited data especially on graphene derivatives such as graphene platelets (GP). In this work, we compared multi-walled carbon nanotubes (MWCNT) and two different types of pristine GP in their potential to activate inflammasome NLRP3 (The nod-like receptor family pyrin domain containing 3) in vitro. Our study is focused on exposure of THP-1/THP1-null cells and peripheral blood monocytes to C-BNM as representative models of canonical and alternative pathways, respectively. Although all nanomaterials were extensively accumulated in the cytoplasm, increasing doses of all C-BNM did not lead to cell death. We observed direct activation of NLRP3 via destabilization of lysosomes and release of cathepsin B into cytoplasm only in the case of MWCNTs. Direct activation of NLRP3 by both GP was statistically insignificant but could be induced by synergic action with muramyl dipeptide (MDP), as a representative molecule of the family of pathogen-associated molecular patterns (PAMPs). This study demonstrates a possible proinflammatory potential of GP and MWCNT acting through NLRP3 activation.

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Carbon-Based Nanomaterials for Delivery of Biologicals and Therapeutics: A Cutting-Edge Technology

C – Journal of Carbon Research ◽

10.3390/c7010019 ◽

2021 ◽

Vol 7 (1) ◽

pp. 19

Author(s):

Alok Mahor ◽

Prem Prakash Singh ◽

Peeyush Bharadwaj ◽

Neeraj Sharma ◽

Surabhi Yadav ◽

...

Keyword(s):

Biomedical Applications ◽

Graphene Quantum Dots ◽

Future Perspectives ◽

Thermal Features ◽

Ancient Times ◽

Characteristic Features ◽

Carbon Based Nanomaterials ◽

Carbon Based

After hydrogen and oxygen, carbon is the third most abundant component present in the cosmos with excellent characteristic features of binding to itself and nearly all elements. Since ancient times, carbon-based materials such as graphite, charcoal, and carbon black have been utilized for writing and drawing materials. As these materials possess excellent chemical, mechanical, electrical, and thermal features, they have been readily engineered into carbon-based nanomaterials (CNMs) such as carbon nanotubes, graphene oxide, graphene quantum dots, nanodiamonds, fullerenes, carbon nano-onions, and so forth. These materials are now widely explored in biomedical applications. Thus, the emergence of CNMs has opened up a gateway for the detection, delivery, and treatment of a multitude of diseases. They are being actively researched for applications within tissue engineering, as vaccine vectors, and for the delivery of therapeutics to the immune system. This review focuses on the recent advances in various types of CNMs, their fabrication techniques, and their application in the delivery of therapeutics both in vitro and in vivo. The review also focuses on the toxicity concern of the CNMs and the possible remedies to tackle the toxicity issues. Concluding remarks emphasize all the CNMs discussed in the review over their possible biomedical applications, while the future perspectives section discusses the approaches to bring CNMs into the mainstream of clinical trials and their therapeutic applications.

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P600 Absolute quantification of myocardial perfusion using real-time myocardial contrast echocardiography: an in vitro study and first in vivo results

European Heart Journal ◽

10.1016/s0195-668x(03)94038-3 ◽

2003 ◽

Vol 24 (5) ◽

pp. 102 ◽

Cited By ~ 1

Author(s):

R VOGEL

Keyword(s):

Myocardial Perfusion ◽

Real Time ◽

Contrast Echocardiography ◽

In Vitro Study ◽

Absolute Quantification ◽

Vitro Study ◽

Myocardial Contrast Echocardiography

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Effects of TSH and calcitriol on bone metabolism: in vivo and in vitro study

Bone Abstracts ◽

10.1530/boneabs.3.pp78 ◽

2014 ◽

Author(s):

Ivo Dumic-Cule ◽

Dunja Rogic ◽

Damir Jezek ◽

Lovorka Grgurevic ◽

Slobodan Vukicevic

Keyword(s):

Bone Metabolism ◽

In Vitro Study ◽

Vitro Study

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Involvement of cytochrome P450 2D in the formation of serotonin in the brain: in vivo and in vitro study

10.26226/morressier.5785edc6d462b80296c9934c ◽

2016 ◽

Author(s):

Anna Haduch

Keyword(s):

Cytochrome P450 ◽

In Vitro Study ◽

Vitro Study ◽

The Brain

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Functional and Transcriptional Adaptations of Blood Monocytes Recruited to the Cystic Fibrosis Airway Microenvironment In Vitro

International Journal of Molecular Sciences ◽

10.3390/ijms22052530 ◽

2021 ◽

Vol 22 (5) ◽

pp. 2530

Author(s):

Bijean D. Ford ◽

Diego Moncada Giraldo ◽

Camilla Margaroli ◽

Vincent D. Giacalone ◽

Milton R. Brown ◽

...

Keyword(s):

Cystic Fibrosis ◽

Bactericidal Activity ◽

Scavenger Receptor ◽

Receptor Expression ◽

Blood Monocytes ◽

Bacterial Killing ◽

Blood Neutrophils ◽

Vitro Model

Cystic fibrosis (CF) lung disease is dominated by the recruitment of myeloid cells (neutrophils and monocytes) from the blood which fail to clear the lung of colonizing microbes. In prior in vitro studies, we showed that blood neutrophils migrated through the well-differentiated lung epithelium into the CF airway fluid supernatant (ASN) mimic the dysfunction of CF airway neutrophils in vivo, including decreased bactericidal activity despite an increased metabolism. Here, we hypothesized that, in a similar manner to neutrophils, blood monocytes undergo significant adaptations upon recruitment to CFASN. To test this hypothesis, primary human blood monocytes were transmigrated in our in vitro model into the ASN from healthy control (HC) or CF subjects to mimic in vivo recruitment to normal or CF airways, respectively. Surface phenotype, metabolic and bacterial killing activities, and transcriptomic profile by RNA sequencing were quantified post-transmigration. Unlike neutrophils, monocytes were not metabolically activated, nor did they show broad differences in activation and scavenger receptor expression upon recruitment to the CFASN compared to HCASN. However, monocytes recruited to CFASN showed decreased bactericidal activity. RNASeq analysis showed strong effects of transmigration on monocyte RNA profile, with differences between CFASN and HCASN conditions, notably in immune signaling, including lower expression in the former of the antimicrobial factor ISG15, defensin-like chemokine CXCL11, and nitric oxide-producing enzyme NOS3. While monocytes undergo qualitatively different adaptations from those seen in neutrophils upon recruitment to the CF airway microenvironment, their bactericidal activity is also dysregulated, which could explain why they also fail to protect CF airways from infection.

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64Cu-ATSM/64Cu-Cl2 and their relationship to hypoxia in glioblastoma: a preclinical study

EJNMMI Research ◽

10.1186/s13550-019-0586-6 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 4

Author(s):

Elodie A. Pérès ◽

Jérôme Toutain ◽

Louis-Paul Paty ◽

Didier Divoux ◽

Méziane Ibazizène ◽

...

Keyword(s):

Ex Vivo ◽

In Vitro Study ◽

Tumor Location ◽

Preclinical Study ◽

Significant Rise ◽

Lower Oxygen ◽

Cu Complexes ◽

Ex Vivo Study

Abstract Background Diacetyl-bis(N4-methylthiosemicarbazone), labeled with 64Cu (64Cu-ATSM) has been suggested as a promising tracer for imaging hypoxia. However, various controversial studies highlighted potential pitfalls that may disable its use as a selective hypoxic marker. They also highlighted that the results may be tumor location dependent. Here, we first analyzed uptake of Cu-ATSM and its less lipophilic counterpart Cu-Cl2 in the tumor over time in an orthotopic glioblastoma model. An in vitro study was also conducted to investigate the hypoxia-dependent copper uptake in tumor cells. We then further performed a comprehensive ex vivo study to compare 64Cu uptake to hypoxic markers, specific cellular reactions, and also transporter expression. Methods μPET was performed 14 days (18F-FMISO), 15 days (64Cu-ATSM and 64Cu-Cl2), and 16 days (64Cu-ATSM and 64Cu-Cl2) after C6 cell inoculation. Thereafter, the brains were withdrawn for further autoradiography and immunohistochemistry. C6 cells were also grown in hypoxic workstation to analyze cellular uptake of Cu complexes in different oxygen levels. Results In vivo results showed that Cu-ASTM and Cu-Cl2 accumulated in hypoxic areas of the tumors. Cu-ATSM also stained, to a lesser extent, non-hypoxic regions, such as regions of astrogliosis, with high expression of copper transporters and in particular DMT-1 and CTR1, and also characterized by the expression of elevated astrogliosis. In vitro results show that 64Cu-ATSM showed an increase in the uptake only in severe hypoxia at 0.5 and 0.2% of oxygen while for 64Cu-Cl2, the cell retention was significantly increased at 5% and 1% of oxygen with no significant rise at lower oxygen percentages. Conclusion In the present study, we show that Cu-complexes undoubtedly accumulate in hypoxic areas of the tumors. This uptake may be the reflection of a direct dependency to a redox metabolism and also a reflection of hypoxic-induced overexpression of transporters. We also show that Cu-ATSM also stained non-hypoxic regions such as astrogliosis.

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5-O-Demethylnobiletin Alleviates CCl4-Induced Acute Liver Injury by Equilibrating ROS-Mediated Apoptosis and Autophagy Induction

International Journal of Molecular Sciences ◽

10.3390/ijms22031083 ◽

2021 ◽

Vol 22 (3) ◽

pp. 1083

Author(s):

Sukkum Ngullie Chang ◽

Se Ho Kim ◽

Debasish Kumar Dey ◽

Seon Min Park ◽

Omaima Nasif ◽

...

Keyword(s):

Liver Injury ◽

Acute Liver Injury ◽

Elevated Serum ◽

In Vitro Study ◽

Biological Properties ◽

Autophagic Flux ◽

Serum Aminotransferase ◽

Ccl4 Treatment

Polymethoxyflavanoids (PMFs) have exhibited a vast array of therapeutic biological properties. 5-O-Demethylnobiletin (5-DN) is one such PMF having anti-inflammatory activity, yet its role in hepatoprotection has not been studied before. Results from in vitro study revealed that 5-DN did not exert a high level of cytotoxicity on HepG2 cells at 40 μM, and it was able to rescue HepG2 cell death induced by carbon tetrachloride (CCl4). Subsequently, we investigated acute liver injury on BALB/c mice induced by CCl4 through the intraperitoneal injection of 1 mL/kg CCl4 and co-administration of 5-DN at (1 and 2 mg/kg) by oral gavage for 15 days. The results illustrated that treatment with 5-DN attenuated CCl4-induced elevated serum aminotransferase (AST)/alanine aminotransferase (ALT) ratio and significantly ameliorated severe hepatic damage such as inflammation and fibrosis evidenced through lesser aberrations in the liver histology of 5-DN dose groups. Additionally, 5-DN efficiently counteracted and equilibrated the production of ROS accelerated by CCl4 and dramatically downregulated the expression of CYP2E1 vitally involved in converting CCl4 to toxic free radicals and also enhanced the antioxidant enzymes. 5-DN treatment also inhibited cell proliferation and inflammatory pathway abnormally regulated by CCl4 treatment. Furthermore, the apoptotic response induced by CCl4 treatment was remarkably reduced by enhanced Bcl-2 expression and noticeable reduction in Bax, Bid, cleaved caspase 3, caspase 9, and apaf-1 expression. 5-DN treatment also induced the conversion of LC3 and promoted the autophagic flux. Conclusively, 5-DN exhibited hepatoprotective effects in vitro and in vivo and prevented liver fibrosis induced by CCl4.

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Effect of cannulation site on emboli travel during cardiac surgery

Journal of Cardiothoracic Surgery ◽

10.1186/s13019-021-01564-1 ◽

2021 ◽

Vol 16 (1) ◽

Author(s):

Mira Puthettu ◽

Stijn Vandenberghe ◽

Stefanos Demertzis

Keyword(s):

Cardiac Surgery ◽

In Vitro Study ◽

Blood Stream ◽

Air Bubbles ◽

Arterial Tree ◽

Air Bubble ◽

Cannulation Site ◽

The Brain

Abstract Background During cardiac surgery, micro-air emboli regularly enter the blood stream and can cause cognitive impairment or stroke. It is not clearly understood whether the most threatening air emboli are generated by the heart-lung machine (HLM) or by the blood-air contact when opening the heart. We performed an in vitro study to assess, for the two sources, air emboli distribution in the arterial tree, especially in the brain region, during cardiac surgery with different cannulation sites. Methods A model of the arterial tree was 3D printed and included in a hydraulic circuit, divided such that flow going to the brain was separated from the rest of the circuit. Air micro-emboli were injected either in the HLM (“ECC Bubbles”) or in the mock left ventricle (“Heart Bubbles”) to simulate the two sources. Emboli distribution was measured with an ultrasonic bubble counter. Five repetitions were performed for each combination of injection site and cannulation site, where air bubble counts and volumes were recorded. Air bubbles were separated in three categories based on size. Results For both injection sites, it was possible to identify statistically significant differences between cannulation sites. For ECC Bubbles, axillary cannulation led to a higher amount of air bubbles in the brain with medium-sized bubbles. For Heart Bubbles, aortic cannulation showed a significantly bigger embolic load in the brain with large bubbles. Conclusions These preliminary in vitro findings showed that air embolic load in the brain may be dependent on the cannulation site, which deserves further in vivo exploration.

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