Electroformation of liposomes and phytosomes using copper electrode

Behuria, Himadri Gourav; Biswal, Bijesh K.; Sahu, Santosh Kumar

doi:10.1080/08982104.2020.1800729

Cited by 8 publications

(5 citation statements)

References 53 publications

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“…The substrate for spreading the lipid film must be electrically conductive. Commonly used substrates include platinum wires 32,41 , ITO coated glasses [42][43][44] , stainless steel wires 45,46 , copper electrodes 47 , and carbon fiber microelectrodes 48 .…”

Section: Introductionmentioning

confidence: 99%

“…32 The substrate for spreading the lipid film must be electrically conductive. Commonly used substrates include platinum wires, 32,41 ITO coated glasses, [42][43][44] stainless steel wires, 45,46 copper electrodes, 47 and carbon fiber microelectrodes. 48 One implicit assumption that is often made in the preparation of GUVs is that these vesicles have the same composition as the lipid mixture that was initially used to grow them.…”

Section: Introductionmentioning

confidence: 99%

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Binding of His-tagged fluorophores to lipid bilayers of giant vesicles

et al. 2022

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Binding of His-tagged fluorophores to lipid bilayers of giant vesicles

et al. 2022

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“… 5 Electroformation of LUVs on the copper electrode produced unilamellar, mono-dispersed vesicles of 0.25–2.0 μ diameter, which are large enough to be visualized at 1500 X magnification and had a vesicle count of ∼3600/mL. 27 ( Figure 3 A,B). Unilamellarity of the vesicles was determined by the calculation of the unilamellarity index ( I U ) that is defined by the following formula where F T = F 0 – F R , F 0 = initial fluorescence, F R = the residual unquenchable fluorescence left after the vesicles were treated with triton X-100.…”

Section: Resultsmentioning

confidence: 99%

“…As amphiphilic compounds exhibit an increased interaction with biological membranes, LUVs were used as a model system for the investigation of their membrane interaction . Electroformation of LUVs on the copper electrode produced unilamellar, mono-dispersed vesicles of 0.25–2.0 μ diameter, which are large enough to be visualized at 1500 X magnification and had a vesicle count of ∼3600/mL . (Figure A,B).…”

Section: Resultsmentioning

confidence: 99%

Lipid Flip-Flop-Inducing Antimicrobial Phytochemicals from Gymnema sylvestre are Bacterial Membrane Permeability Enhancers

et al. 2021

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An amphiphilic phytochemical fraction isolated from methanol extract of Gymnema sylvestre leaf powder contained six terpenoids, two flavonoids, and one alkaloid that induced rapid flip-flop of fluorescent phospholipid analog in the phosphatidyl choline bilayer. Lipid-flipping activity of the methanol-extracted fraction of G. sylvestre (MEFGS) was dose-dependent and time-dependent with a rate constant k = (12.09 ± 0.94) mg –1 min –1 that was saturable at (40 ± 1) % flipping of the fluorescent lipid analogue. Interactions of MEFGS phytochemicals with large unilamelar vesicles led to time-dependent change in their rounded morphology into irregular shapes, indicating their membrane-destabilizing activity. MEFGS exhibited antibacterial activity on Escherichia coli (MTCC-118), Staphylococcus aureus (MTCC-212), and Pseudomonas aeruginosa (MTCC-1035) with IC 50 values 0.5, 0.35, and 0.1 mg/mL, respectively. Phytochemicals in MEFGS increased membrane permeabilization in all three bacteria, as indicated by 23, 17, and 17% increase in the uptake of crystal violet, respectively. MEFGS enhanced membrane damage, resulting in a 3–5 fold increase in leakage of cytosolic ions, 0.5–2 fold increase in leakage of PO 4 – , and 15–20% increase in loss of cellular proteins. MEFGS synergistically increased the efficacy of curcumin, amoxillin, ampicillin, and cefotaxime on S. aureus probably by enhancing their permeability into the bacterium. For the first time, our study reveals that phytochemicals from G. sylvestre enhance the permeability of the bacterial plasma membrane by facilitating flip-flop of membrane lipids. Lipid-flipping phytochemicals from G. sylvestre can be used as adjuvant therapeutics to enhance the efficacy of antibacterials by increasing their bioavailability in the target bacteria.

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“…Electrode replacement or annealing is recommended after approximately five experiments [38]. Steel syringes and copper electrodes have also been suggested as cost-friendly alternatives [39,40]. Furthermore, titanium electrodes have been advocated by some groups since their usage seems to decrease lipid peroxidation when compared to ITO electrodes [11,[41][42][43].…”

Section: Electrode Materials and Cleaningmentioning

confidence: 99%

Giant Unilamellar Vesicle Electroformation: What to Use, What to Avoid, and How to Quantify the Results

et al. 2021

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Since its inception more than thirty years ago, electroformation has become the most commonly used method for growing giant unilamellar vesicles (GUVs). Although the method seems quite straightforward at first, researchers must consider the interplay of a large number of parameters, different lipid compositions, and internal solutions in order to avoid artifactual results or reproducibility problems. These issues motivated us to write a short review of the most recent methodological developments and possible pitfalls. Additionally, since traditional manual analysis can lead to biased results, we have included a discussion on methods for automatic analysis of GUVs. Finally, we discuss possible improvements in the preparation of GUVs containing high cholesterol contents in order to avoid the formation of artifactual cholesterol crystals. We intend this review to be a reference for those trying to decide what parameters to use as well as an overview providing insight into problems not yet addressed or solved.

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Electroformation of liposomes and phytosomes using copper electrode

Cited by 8 publications

References 53 publications

Binding of His-tagged fluorophores to lipid bilayers of giant vesicles

Binding of His-tagged fluorophores to lipid bilayers of giant vesicles

Lipid Flip-Flop-Inducing Antimicrobial Phytochemicals from Gymnema sylvestre are Bacterial Membrane Permeability Enhancers

Giant Unilamellar Vesicle Electroformation: What to Use, What to Avoid, and How to Quantify the Results

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