Production of giant unilamellar vesicles and encapsulation of lyotropic nematic liquid crystals

Bao, Peng; Paterson, Daniel A.; Peyman, Sally A.; Jones, John C.; Sandoe, Jonathan; Gleeson, Helen F.; Evans, Stephen D.; Bushby, Richard J.

doi:10.1039/d0sm01684e

Cited by 21 publications

(14 citation statements)

References 64 publications

(85 reference statements)

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“…10,11,36 The oil phase containing a mixture of hexane and chloroform is incompatible with the PDMS chip, and our attempt to use n-octanol as the oil phase failed to generate liposomes, as it was recently shown that physical perturbation is needed to trigger dewetting for such a system. 11,38 Using a mixture of paraffin oil and n-hexanol (v/v% of 40 : 60), our results show that double emulsions with varying sizes can be stably produced, leading to liposome formation via spontaneous dewetting. At this stage, the detailed mechanism of how the mixture of paraffin oil and n-hexanol can trigger dewetting process is still unclear, as a more comprehensive analysis, such as comparing the interfacial tensions between three phases, is required to reveal it.…”

Section: Formation Of Double-emulsion Droplets and Liposomes With Hig...mentioning

confidence: 82%

Chip assisted formation of phase-separated liposomes for reconstituting spatial protein–lipid interactions

et al. 2022

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Section: Formation Of Double-emulsion Droplets and Liposomes With Hig...mentioning

confidence: 82%

Chip assisted formation of phase-separated liposomes for reconstituting spatial protein–lipid interactions

et al. 2022

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“…To verify the formation of lipid membranes, we encapsulated fluorescein and 0.2 M sucrose in the inner core of the double emulsions (along with 2% w/w PVA) prior to tip streaming as previously described. , The resulting droplets were collected in a 2% w/w PVA solution (blank) or in 2% w/w PVA solution containing 50 μg/mL α-hemolysin. The latter spontaneously incorporates into the phospholipid membrane, forming narrow pores.…”

Section: Resultsmentioning

confidence: 99%

Tip Streaming of a Lipid-Stabilized Double Emulsion Generated in a Microfluidic Channel

et al. 2021

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Water/oil/water (w/o/w) double emulsions (DEs) are multicompartment structures which can be used in many technological applications and in fundamental studies as models of cells like microreactors or templates for other materials. Herein we study the flow dynamics of water/oil/water double emulsions generated in a microfluidic device and stabilized with the phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). We show that by varying the concentration of lipids in the oil phase (chloroform), or by modulating the viscosity of the aqueous continuous phase, the double emulsions under flow exhibit a rich dynamic behavior. An initial deformation of the double emulsions is followed by tube extraction at the rear end, relative to the flow direction, resulting in pinch off at the tube extremity, by which small aqueous compartments are released. These compartments are phospholipid vesicles as deduced from fluorescence

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“…Concerning octanol de-wetting and separation on-chip demonstrated by Deshpande et al [86], LO phase concentration was adjusted to provide an appropriate interfacial tension that induces spontaneously de-wetting. This assures the separation of octanol residues from GUVs in a two-phase sedimentation process that takes about 1-2 h. This principle of separation by sedimentation is due to the octanol buoyancy that allows its precipitation for further supernatant extraction off-chip [87], which makes the proposed device a simpler low-cost alternative with high effectiveness for the synthesis of GUVs.…”

Section: Low-cost Octanol-assisted Liposomes Assemblymentioning

confidence: 99%

Design and Manufacture of a Low-Cost Microfluidic System for the Synthesis of Giant Liposomes for the Encapsulation of Yeast Homologues: Applications in the Screening of Membrane-Active Peptide Libraries

et al. 2021

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The discovery of new membrane-active peptides (MAPs) is an area of considerable interest in modern biotechnology considering their ample applicability in several fields ranging from the development of novel delivery vehicles (via cell-penetrating peptides) to responding to the latent threat of antibiotic resistance (via antimicrobial peptides). Different strategies have been devised for such discovery process, however, most of them involve costly, tedious, and low-efficiency methods. We have recently proposed an alternative route based on constructing a non-rationally designed library recombinantly expressed on the yeasts’ surfaces. However, a major challenge is to conduct a robust and high-throughput screening of possible candidates with membrane activity. Here, we addressed this issue by putting forward low-cost microfluidic platforms for both the synthesis of Giant Unilamellar Vesicles (GUVs) as mimicking entities of cell membranes and for providing intimate contact between GUVs and homologues of yeasts expressing MAPs. The homologues were chitosan microparticles functionalized with the membrane translocating peptide Buforin II, while intimate contact was through passive micromixers with different channel geometries. Both microfluidic platforms were evaluated both in silico (via Multiphysics simulations) and in vitro with a high agreement between the two approaches. Large and stable GUVs (5–100 µm) were synthesized effectively, and the mixing processes were comprehensively studied leading to finding the best operating parameters. A serpentine micromixer equipped with circular features showed the highest average encapsulation efficiencies, which was explained by the unique mixing patterns achieved within the device. The microfluidic devices developed here demonstrate high potential as platforms for the discovery of novel MAPs as well as for other applications in the biomedical field such as the encapsulation and controlled delivery of bioactive compounds.

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Production of giant unilamellar vesicles and encapsulation of lyotropic nematic liquid crystals

Abstract: We describe a modified microfluidic method for making Giant Unilamellar Vesicles (GUVs) via water/octanol-lipid/water double emulsion droplets and encapsulation of nematic lyotropic liquid crystals (LNLCs).

Cited by 21 publications

References 64 publications

Chip assisted formation of phase-separated liposomes for reconstituting spatial protein–lipid interactions

Chip assisted formation of phase-separated liposomes for reconstituting spatial protein–lipid interactions

Tip Streaming of a Lipid-Stabilized Double Emulsion Generated in a Microfluidic Channel

Design and Manufacture of a Low-Cost Microfluidic System for the Synthesis of Giant Liposomes for the Encapsulation of Yeast Homologues: Applications in the Screening of Membrane-Active Peptide Libraries

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