Nile Red Quantifier: a novel and quantitative tool to study lipid accumulation in patient-derived circulating monocytes using confocal microscopy

Schnitzler, Johan G.; Moens, Sophie J. Bernelot; Tiessens, Feiko; Bakker, Guido J.; Dallinga‐Thie, Geesje M.; Groen, Albert K.; Nieuwdorp, Max; Stroes, Erik S.G.; Kroon, Jeffrey

doi:10.1194/jlr.d073197

Cited by 24 publications

(23 citation statements)

References 43 publications

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“…Murine peripheral blood mononuclear cells (PBMCs) were obtained from whole blood samples through density centrifugation using Lymphoprep (Stemcell Technologies, Koln, Germany; D = 1.077 g/mL) as described in detail previously ( Schnitzler et al, 2017 ). In short, blood was diluted in a 1:1 ratio with PBS enriched with 2 mM EDTA and subsequently added to a layer of Lymphoprep.…”

Section: Methodsmentioning

confidence: 99%

Inhibition of PFKFB3 Hampers the Progression of Atherosclerosis and Promotes Plaque Stability

Poels

Schnitzler

Waissi

et al. 2020

Front. Cell Dev. Biol.

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Aims 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase (PFKFB)3-mediated glycolysis is pivotal in driving macrophage- and endothelial cell activation and thereby inflammation. Once activated, these cells play a crucial role in the progression of atherosclerosis. Here, we analyzed the expression of PFKFB3 in human atherosclerotic lesions and investigated the therapeutic potential of pharmacological inhibition of PFKFB3 in experimental atherosclerosis by using the glycolytic inhibitor PFK158. Methods and Results PFKFB3 expression was higher in vulnerable human atheromatous carotid plaques when compared to stable fibrous plaques and predominantly expressed in plaque macrophages and endothelial cells. Analysis of advanced plaques of human coronary arteries revealed a positive correlation of PFKFB3 expression with necrotic core area. To further investigate the role of PFKFB3 in atherosclerotic disease progression, we treated 6–8 weeks old male Ldlr –/– mice. These mice were fed a high cholesterol diet for 13 weeks, of which they were treated for 5 weeks with the glycolytic inhibitor PFK158 to block PFKFB3 activity. The incidence of fibrous cap atheroma (advanced plaques) was reduced in PFK158-treated mice. Plaque phenotype altered markedly as both necrotic core area and intraplaque apoptosis decreased. This coincided with thickening of the fibrous cap and increased plaque stability after PFK158 treatment. Concomitantly, we observed a decrease in glycolysis in peripheral blood mononuclear cells compared to the untreated group, which alludes that changes in the intracellular metabolism of monocyte and macrophages is advantageous for plaque stabilization. Conclusion High PFKFB3 expression is associated with vulnerable atheromatous human carotid and coronary plaques. In mice, high PFKFB3 expression is also associated with a vulnerable plaque phenotype, whereas inhibition of PFKFB3 activity leads to plaque stabilization. This data implies that inhibition of inducible glycolysis may reduce inflammation, which has the ability to subsequently attenuate atherogenesis.

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Section: Methodsmentioning

confidence: 99%

Inhibition of PFKFB3 Hampers the Progression of Atherosclerosis and Promotes Plaque Stability

Poels

Schnitzler

Waissi

et al. 2020

Front. Cell Dev. Biol.

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“…Since lipid uptake by monocytes induces inflammation and this is directly connected to an elevated LD content, Nile Red served as a platform in the development of a quantitative imaging-based analysis of lipid accumulation [ 66 ]. Recently BioTek ® has developed a phenotypic-based assay using lipophilic dyes (Nile Red and BODIPY 493/503) to probe LDs formation in hepatic HepG2 cells associated with non-alcoholic fatty liver disease (NAFLD) caused by fatty acids overload [ 67 ].…”

Section: Application Of Fluorescent Ld Probesmentioning

confidence: 99%

“…Recently BioTek ® has developed a phenotypic-based assay using lipophilic dyes (Nile Red and BODIPY 493/503) to probe LDs formation in hepatic HepG2 cells associated with non-alcoholic fatty liver disease (NAFLD) caused by fatty acids overload [ 67 ]. However, Nile Red was also able to detect alterations in neutral LD content in circulating monocytes derived from patients [ 66 ] and report LDs NAFLD-associated overproduction [ 67 ].…”

Section: Application Of Fluorescent Ld Probesmentioning

confidence: 99%

Recent Advances in Fluorescent Probes for Lipid Droplets

2018

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Lipid droplets (LDs) are organelles that serve as the storage of intracellular neutral lipids. LDs regulate many physiological processes. They recently attracted attention after extensive studies showed their involvement in metabolic disorders and diseases such as obesity, diabetes, and cancer. Therefore, it is of the highest importance to have reliable imaging tools. In this review, we focus on recent advances in the development of selective fluorescent probes for LDs. Their photophysical properties are described, and their advantages and drawbacks in fluorescence imaging are discussed. At last, we review the reported applications using these probes including two-photon excitation, in vivo and tissue imaging, as well as LDs tracking.

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“…Nile red is a lipophilic dye and has been used to stain lipids, cells, and oils [50,51,52,53]. In this study, Nile red was employed to stain vegetable oils to evaluate oil encapsulation efficiency.…”

Section: Resultsmentioning

confidence: 99%

Ultrasound-Assisted Encapsulation of Sacha Inchi (Plukenetia volubilis Linneo.) Oil in Alginate-Chitosan Nanoparticles

et al. 2019

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Sacha inchi oil is rich in essential and non-essential fatty acids and other types of bioactive agents like tocopherols and polyphenolic compounds, which are very well-known antioxidants. In this study, the encapsulation of sacha inchi oil in alginate (AL) and chitosan (CS) nanoparticles was achieved with the assistance of high-intensity ultrasound. Nanoemulsion is the most effective delivery and high stability system for lipophilic bioactive agents. Chitosan and surfactant concentrations were varied to study their effect on particle formulations. Size, zeta-potential, polydispersity, and stability of particles were determined in time to optimize the preparation conditions. Sacha inchi oil encapsulated in AL-CS nanoparticles showed a higher loading efficiency and stability for short and long periods compared with other vegetable oils such as olive and soybean. Also, because of the types of tocopherols present in sacha inchi oil (γ- and δ-tocopherols), a much higher antioxidant activity (95% of radical reduction in 15 min) was found in comparison with nanocapsules with olive oil, which contain α-tocopherols. The particles showed high efficiency of protein loading at high concentration of bovine serum albumin (BSA) and a low rate of leaching profiles in various testing media like simulated gastric and intestinal fluids with/without enzymes, that is, pepsin 0.1% (w/v) and pancreatin 0.1% (w/v), respectively.

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Nile Red Quantifier: a novel and quantitative tool to study lipid accumulation in patient-derived circulating monocytes using confocal microscopy

Cited by 24 publications

References 43 publications

Inhibition of PFKFB3 Hampers the Progression of Atherosclerosis and Promotes Plaque Stability

Inhibition of PFKFB3 Hampers the Progression of Atherosclerosis and Promotes Plaque Stability

Recent Advances in Fluorescent Probes for Lipid Droplets

Ultrasound-Assisted Encapsulation of Sacha Inchi (Plukenetia volubilis Linneo.) Oil in Alginate-Chitosan Nanoparticles

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