Size fractionation and size characterization of nanoemulsions of lipid droplets and large unilamellar lipid vesicles by asymmetric-flow field-flow fractionation/multi-angle light scattering and dynamic light scattering

Vezočnik, Valerija; Rebolj, Katja; Sitar, Simona; Ota, Katja; Tušek‐Žnidarič, Magda; Štrus, Jasna; Sepčić, Kristina; Pahovnik, David; Maček, Peter; Žagar, Ema

doi:10.1016/j.chroma.2015.09.048

Cited by 27 publications

(26 citation statements)

References 51 publications

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“…The methodology applied in this study for NC and NE preparation, AFM analyses and previous studies indicate that the NC are spherical. For spherical particles R g /R h of 0.775 is characteristic of spheres with uniform density, such as polystyrene latexes (Brewer and Striegel, 2009), values close to 1.0 are obtained with vesicles or hollow spheres (Stauch et al, 2002;Vezočnik et al, 2015), values above 1 are obtained with elongated or porous structures and values as low as 0.3 are characteristic of microgels (Kunz et al, 1983). These values are in agreement with literature values for similar types of NC (Roy et al, 2015) and liposomes (Vezočnik et al, 2015).…”

Section: Accepted Manuscriptsupporting

confidence: 89%

“…For spherical particles R g /R h of 0.775 is characteristic of spheres with uniform density, such as polystyrene latexes (Brewer and Striegel, 2009), values close to 1.0 are obtained with vesicles or hollow spheres (Stauch et al, 2002;Vezočnik et al, 2015), values above 1 are obtained with elongated or porous structures and values as low as 0.3 are characteristic of microgels (Kunz et al, 1983). These values are in agreement with literature values for similar types of NC (Roy et al, 2015) and liposomes (Vezočnik et al, 2015). The higher R g /R h for PLA-PEG NC may be attributed to the PEG corona creating a dense hydration layer around the oil-polymer vesicle…”

Section: Accepted Manuscriptmentioning

confidence: 97%

See 1 more Smart Citation

Impact of dose and surface features on plasmatic and liver concentrations of biodegradable polymeric nanocapsules

Oliveira

Paula

Roatt

et al. 2017

European Journal of Pharmaceutical Sciences

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Section: Accepted Manuscriptsupporting

confidence: 89%

Section: Accepted Manuscriptmentioning

confidence: 97%

Impact of dose and surface features on plasmatic and liver concentrations of biodegradable polymeric nanocapsules

Oliveira

Paula

Roatt

et al. 2017

European Journal of Pharmaceutical Sciences

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“…Dynamic light scattering can provide quantitative information on particle size and shape, with relatively fast measurements ( Faustino et al, 2014 ). However, for heterogeneous and highly polydisperse systems, the results can be inaccurate ( Vezočnik et al, 2015 ). If the LDs sample in study is highly heterogeneous, which may naturally occur, the light scattered from larger LDs may obscure the light scattered from the smaller ones.…”

Section: Methods For Lipid Droplets Characterizationmentioning

confidence: 99%

Methods for Lipid Droplet Biophysical Characterization in Flaviviridae Infections

Martins

Santos

2018

Front. Microbiol.

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Lipid droplets (LDs) are intracellular organelles for neutral lipid storage, originated from the endoplasmic reticulum. They play an essential role in lipid metabolism and cellular homeostasis. In fact, LDs are complex organelles, involved in many more cellular processes than those initially proposed. They have been extensively studied in the context of LD-associated pathologies. In particular, LDs have emerged as critical for virus replication and assembly. Viruses from the Flaviviridae family, namely dengue virus (DENV), hepatitis C virus (HCV), West Nile virus (WNV), and Zika virus (ZIKV), interact with LDs to usurp the host lipid metabolism for their own viral replication and pathogenesis. In general, during Flaviviridae infections it is observed an increasing number of host intracellular LDs. Several viral proteins interact with LDs during different steps of the viral life cycle. The HCV core protein and DENV capsid protein, extensively interact with LDs to regulate their replication and assembly. Detailed studies of LDs in viral infections may contribute for the development of possible inhibitors of key steps of viral replication. Here, we reviewed different techniques that can be used to characterize LDs isolated from infected or non-infected cells. Microscopy studies have been commonly used to observe LDs accumulation and localization in infected cell cultures. Fluorescent dyes, which may affect LDs directly, are widely used to probe LDs but there are also approaches that do not require the use of fluorescence, namely stimulated Raman scattering, electron and atomic force microscopy-based approaches. These three are powerful techniques to characterize LDs morphology. Raman scattering microscopy allows studying LDs in a single cell. Electron and atomic force microscopies enable a better characterization of LDs in terms of structure and interaction with other organelles. Other biophysical techniques, such as dynamic light scattering and zeta potential are also excellent to characterize LDs in terms of size in a simple and fast way and test possible LDs interaction with viral proteins. These methodologies are reviewed in detail, in the context of viral studies.

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“…MALS represents a static light scattering technique. 15 , 16 Depending on the analysis model, MALS determines, using the angular dependence of the time-averaged scattering intensity, the geometric radius ( R geo ) or the root-mean-square radius, commonly known as the radius of gyration ( R g ).…”

Section: Introductionmentioning

confidence: 99%

Multiple Techniques for Size Determination of Generalized Modules for Membrane Antigens from Salmonella typhimurium and Salmonella enteritidis

et al. 2017

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In the last years, outer membrane vesicles have attracted a lot of attention for the development of vaccines against bacterial pathogens. Extracellular vesicles can be obtained in high yields by genetic mutations, resulting in generalized modules for membrane antigens (GMMA). Methods to check the quality, consistency of production, and stability of GMMA vaccines are of fundamental importance. In this context, analytical methods for size distribution determination and verifying the integrity and possible aggregation of GMMA particles are strongly needed. Herein, GMMA particle size distribution has been evaluated by means of three different techniques. Dynamic light scattering (DLS), multiangle light scattering (MALS) coupled with high-performance liquid chromatography–size exclusion chromatography (SEC), and nanoparticle tracking analysis (NTA) have been compared to characterize GMMA from different mutants of Salmonella typhimurium and Salmonella enteritidis strains. We found that the presence of O-antigen chains on GMMA determined higher Z-average diameters by DLS compared to size estimation by MALS and that the hydrodynamic diameter increased with the number of O-antigen chains per GMMA particle. In the case of SEC-MALS, the size of the whole population better reflects the size of the most abundant particles, whereas DLS diameter is more influenced by the presence of larger particles in the sample. SEC-MALS and NTA are preferable to DLS for the analysis of bimodal samples, as they better distinguish populations of different size. MALS coupled to a size exclusion chromatography module also allows checking the purity of GMMA preparations, allowing determination of generally occurring contaminants such as soluble proteins and DNA. NTA permits real-time visualization with simultaneous tracking and counting of individual particles, but it is deeply dependent on the choice of data analysis parameters. All of the three techniques have provided complementary information leading to a more complete characterization of GMMA particles.

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Size fractionation and size characterization of nanoemulsions of lipid droplets and large unilamellar lipid vesicles by asymmetric-flow field-flow fractionation/multi-angle light scattering and dynamic light scattering

Cited by 27 publications

References 51 publications

Impact of dose and surface features on plasmatic and liver concentrations of biodegradable polymeric nanocapsules

Impact of dose and surface features on plasmatic and liver concentrations of biodegradable polymeric nanocapsules

Methods for Lipid Droplet Biophysical Characterization in Flaviviridae Infections

Multiple Techniques for Size Determination of Generalized Modules for Membrane Antigens from Salmonella typhimurium and Salmonella enteritidis

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