A new approach for the separation, characterization and testing of potential prionoid protein aggregates through hollow-fiber flow field-flow fractionation and multi-angle light scattering

Marassi, Valentina; Beretti, Francesca; Alessandrini, Andrea; Facci, Paolo; Maraldi, Tullia; Zattoni, Andrea; Reschiglian, Pierluigi; Portolani, Marinella

doi:10.1016/j.aca.2019.08.003

Cited by 19 publications

(16 citation statements)

References 36 publications

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“…The micro-volume variant of FFF, hollow-fiber flow-field flow fractionation (HF5), was demonstrated to be capable of achieving a high performance and low dilution at the same time for the analysis of particles of different natures; this method also allows for applications where a disposable device is needed to avoid cross-contamination [43][44][45][46][47][48][49][50][51].…”

Section: Introductionmentioning

confidence: 99%

Synthesis Monitoring, Characterization and Cleanup of Ag-Polydopamine Nanoparticles Used as Antibacterial Agents with Field-Flow Fractionation

et al. 2022

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Advances in nanotechnology have opened up new horizons in nanomedicine through the synthesis of new composite nanomaterials able to tackle the growing drug resistance in bacterial strains. Among these, nanosilver antimicrobials sow promise for use in the treatment of bacterial infections. The use of polydopamine (PDA) as a biocompatible carrier for nanosilver is appealing; however, the synthesis and functionalization steps used to obtain Ag-PDA nanoparticles (NPs) are complex and require time-consuming cleanup processes. Post-synthesis treatment can also hinder the stability and applicability of the material, and dry, offline characterization is time-consuming and unrepresentative of real conditions. The optimization of Ag-PDA preparation and purification together with well-defined characterization are fundamental goals for the safe development of these new nanomaterials. In this paper, we show the use of field-flow fractionation with multi-angle light scattering and spectrophotometric detection to improve the synthesis and quality control of the production of Ag-PDA NPs. An ad hoc method was able to monitor particle growth in a TLC-like fashion; characterize the species obtained; and provide purified, isolated Ag-PDA nanoparticles, which proved to be biologically active as antibacterial agents, while achieving a short analysis time and being based on the use of green, cost-effective carriers such as water.

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Section: Introductionmentioning

confidence: 99%

Synthesis Monitoring, Characterization and Cleanup of Ag-Polydopamine Nanoparticles Used as Antibacterial Agents with Field-Flow Fractionation

et al. 2022

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“…Field flow fractionation (FFF) is a flow-based sorting method applicable for separation of particles ranging from a few nanometers to about 100 μm in diameter. − This technique has been widely utilized to isolate and characterize biological and nonbiological particles. − To separate particles, a liquid suspension containing particles traveling (laminar flow with a parabolic profile) in a narrow channel (height of 50–500 μm) while being subjected to an externally applied field (liquid flows, centrifugal forces, gravity or electrical fields) perpendicular to the direction of suspension flow (Figure A). − The separation mechanism is based on the differences in density and hydrodynamic properties of particles that define particle mobility under the effect of forces perpendicular to each other; the forward force exerted by laminar channel flow and vertical forces exerted by the externally applied field and diffusion. External field forces drive the accumulation of particles at the bottom wall of the channel, whereas Brownian motion of the particles results in their diffusion toward the channel center. − Depending on their equilibrium position from the wall of the microchannel, particles travel at different speeds and thereby are separated as they move down the microchannel (Figure A).…”

Section: Label-free Microfluidic Methods For Exosome Isolationmentioning

confidence: 99%

Label-Free Isolation of Exosomes Using Microfluidic Technologies

2021

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Exosomes are cell-derived structures packaged with lipids, proteins, and nucleic acids. They exist in diverse bodily fluids and are involved in physiological and pathological processes. Although their potential for clinical application as diagnostic and therapeutic tools has been revealed, a huge bottleneck impeding the development of applications in the rapidly burgeoning field of exosome research is an inability to efficiently isolate pure exosomes from other unwanted components present in bodily fluids. To date, several approaches have been proposed and investigated for exosome separation, with the leading candidate being microfluidic technology due to its relative simplicity, costeffectiveness, precise and fast processing at the microscale, and amenability to automation. Notably, avoiding the need for exosome labeling represents a significant advance in terms of process simplicity, time, and cost as well as protecting the biological activities of exosomes. Despite the exciting progress in microfluidic strategies for exosome isolation and the countless benefits of label-free approaches for clinical applications, current microfluidic platforms for isolation of exosomes are still facing a series of problems and challenges that prevent their use for clinical sample processing. This review focuses on the recent microfluidic platforms developed for labelfree isolation of exosomes including those based on sieving, deterministic lateral displacement, field flow, and pinched flow fractionation as well as viscoelastic, acoustic, inertial, electrical, and centrifugal forces. Further, we discuss advantages and disadvantages of these strategies with highlights of current challenges and outlook of label-free microfluidics toward the clinical utility of exosomes.

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“…At the end of each experiment, 0.5 mg/ml MTT was added and incubated for 3 h at 37°C. After incubation, the medium was removed and acidified isopropanol was added to solubilize the formazan salts [21]. The absorbance was measured at 570 nm using a microplate spectrophotometer (Appliskan, Thermo-Fisher Scientific, Vantaa, Finland).…”

Section: Preparation Of Primary Cultured Cortical Neuronsmentioning

confidence: 99%

Oxidative Stress in Alzheimer’s Disease: In Vitro Therapeutic Effect of Amniotic Fluid Stem Cells Extracellular Vesicles

Gatti

Zavatti

Beretti

et al. 2020

Oxidative Medicine and Cellular Longevity

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Alzheimer’s disease (AD) is characterized by abnormal protein aggregation, deposition of extracellular β-amyloid proteins (Aβ), besides an increase of oxidative stress. Amniotic fluid stem cells (AFSCs) should have a therapeutic potential for neurodegenerative disorders, mainly through a paracrine effect mediated by extracellular vesicles (EV). Here, we examined the effect of EV derived from human AFSCs (AFSC-EV) on the disease phenotypes in an AD neuron primary culture. We observed a positive effect of AFSC-EV on neuron morphology, viability, and Aβ and phospho-Tau levels. This could be due to the apoptotic and autophagic pathway modulation derived from the decrease in oxidative stress. Indeed, reactive oxygen species (ROS) were reduced, while GSH levels were enhanced. This modulation could be ascribed to the presence of ROS regulating enzymes, such as SOD1 present into the AFSC-EV themselves. This study describes the ROS-modulating effects of extracellular vesicles alone, apart from their deriving stem cell, in an AD in vitro model, proposing AFSC-EV as a therapeutic tool to stop the progression of AD.

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A new approach for the separation, characterization and testing of potential prionoid protein aggregates through hollow-fiber flow field-flow fractionation and multi-angle light scattering

Cited by 19 publications

References 36 publications

Synthesis Monitoring, Characterization and Cleanup of Ag-Polydopamine Nanoparticles Used as Antibacterial Agents with Field-Flow Fractionation

Synthesis Monitoring, Characterization and Cleanup of Ag-Polydopamine Nanoparticles Used as Antibacterial Agents with Field-Flow Fractionation

Label-Free Isolation of Exosomes Using Microfluidic Technologies

Oxidative Stress in Alzheimer’s Disease: In Vitro Therapeutic Effect of Amniotic Fluid Stem Cells Extracellular Vesicles

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