Nanoparticle Tracking Analysis of β-Casein Nanocarriers

Accurate physico-chemical characterization of exosomes and liposomes in biological media is challenging due to the inherent complexity of the sample matrix. An appropriate purification step can significantly reduce matrix interferences, and thus facilitate analysis of such demanding samples. Electrical Asymmetrical Flow Field-Flow Fractionation (EAF4) provides online sample purification while simultaneously enabling access to size and Zeta potential of sample constituents in the size range of approx. 1–1000 nm. Hyphenation of EAF4 with Multi-Angle Light Scattering (MALS) and Nanoparticle Tracking Analysis (NTA) detection adds high resolution size and number concentration information turning this setup into a powerful analytical platform for the comprehensive physico-chemical characterization of such challenging samples. We here present EAF4-MALS hyphenated with NTA for the analysis of liposomes and exosomes in complex, biological media. Coupling of the two systems was realized using a flow splitter to deliver the sample at an appropriate flow speed for the NTA measurement. After a proof-of-concept study using polystyrene nanoparticles, the combined setup was successfully applied to analyze liposomes and exosomes spiked into cell culture medium and rabbit serum, respectively. Obtained results highlight the benefits of the EAF4-MALS-NTA platform to study the behavior of these promising drug delivery vesicles under in vivo like conditions.

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Analysis of Neat Biofluids Obtained During Cardiac Surgery Using Nanoparticle Tracking Analysis: Methodological Considerations

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2020.00367 ◽

2020 ◽

Vol 8 ◽

Author(s):

Andrew I. U. Shearn ◽

Sezin Aday ◽

Soumaya Ben-Aicha ◽

Pauline Carnell-Morris ◽

Agnieszka Siupa ◽

...

Keyword(s):

Cardiac Surgery ◽

Nanoparticle Tracking Analysis ◽

Methodological Considerations

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Applying Nanoparticle Tracking Analysis to Characterize the Polydispersity of Aggregates Resulting from Tannin–Polysaccharide Interactions in Wine-Like Media

Molecules ◽

10.3390/molecules24112100 ◽

2019 ◽

Vol 24 (11) ◽

pp. 2100 ◽

Cited By ~ 5

Author(s):

Sijing Li ◽

Kerry L. Wilkinson ◽

Agnieszka Mierczynska-Vasilev ◽

Keren A. Bindon

Keyword(s):

Particle Size ◽

Light Scattering ◽

Weak Interactions ◽

Grape Seed ◽

Alcohol Concentration ◽

Nanoparticle Tracking Analysis ◽

Promising Tool ◽

Light Scattering Intensity ◽

Tannin Concentration ◽

Uv Visible

Interactions between grape seed tannin and either a mannoprotein or an arabinogalactan in model wine solutions of different ethanol concentrations were characterized with nanoparticle tracking analysis (NTA), UV-visible spectroscopy and dynamic light scattering (DLS). NTA results reflected a shift in particle size distribution due to aggregation. Furthermore, the light scattering intensity of each tracked particle measured by NTA demonstrated the presence of aggregates, even when a shift in particle size was not apparent. Mannoprotein and arabinogalactan behaved differently when combined with seed tannin. Mannoprotein formed large, highly light-scattering aggregates, while arabinogalactan exhibited only weak interactions with seed tannin. A 3% difference in alcohol concentration of the model solution (12 vs. 15% v/v) was sufficient to affect the interactions between mannoprotein and tannin when the tannin concentration was high. In summary, this study showed that NTA is a promising tool for measuring polydisperse samples of grape and wine macromolecules, and their aggregates under wine-like conditions. The implications for wine colloidal properties are discussed based on these results.

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Oxidative Stress Product, 4-Hydroxy-2-Nonenal, Induces the Release of Tissue Factor-Positive Microvesicles From Perivascular Cells Into Circulation

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvbaha.120.315187 ◽

2020 ◽

Cited By ~ 1

Author(s):

Shabbir A. Ansari ◽

Shiva Keshava ◽

Usha R. Pendurthi ◽

L. Vijaya Mohan Rao

Keyword(s):

Tissue Factor ◽

Thrombin Generation ◽

Smooth Muscle Actin ◽

Blue Dye ◽

Nanoparticle Tracking Analysis ◽

Perivascular Cells ◽

Aberrant Expression ◽

Intravascular Coagulation ◽

Coagulation Assays ◽

Cellular Source

Objective: TF (Tissue factor) plays a key role in hemostasis, but an aberrant expression of TF leads to thrombosis. The objective of the present study is to investigate the effect of 4-hydroxy-2-nonenal (HNE), the most stable and major oxidant produced in various disease conditions, on the release of TF + microvesicles into the circulation, identify the source of TF + microvesicles origin, and assess their effect on intravascular coagulation and inflammation. Approach and Results: C57BL/6J mice were administered with HNE intraperitoneally, and the release of TF + microvesicles into circulation was evaluated using coagulation assays and nanoparticle tracking analysis. Various cell-specific markers were used to identify the cellular source of TF + microvesicles. Vascular permeability was analyzed by the extravasation of Evans blue dye or fluorescein dextran. HNE administration to mice markedly increased the levels of TF + microvesicles and thrombin generation in the circulation. HNE administration also increased the number of neutrophils in the lungs and elevated the levels of inflammatory cytokines in plasma. Administration of an anti-TF antibody blocked not only HNE-induced thrombin generation but also HNE-induced inflammation. Confocal microscopy and immunoblotting studies showed that HNE does not induce TF expression either in vascular endothelium or circulating monocytes. Microvesicles harvested from HNE-administered mice stained positively with CD248 and α-smooth muscle actin, the markers that are specific to perivascular cells. HNE was found to destabilize endothelial cell barrier integrity. Conclusions: HNE promotes the release of TF + microvesicles from perivascular cells into the circulation. HNE-induced increased TF activity contributes to intravascular coagulation and inflammation.

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