Dataset Information

Imaging of photoacoustic-mediated permeabilization of giant unilamellar vesicles (GUVs).

ABSTRACT: Target delivery of large foreign materials to cells requires transient permeabilization of the cell membrane without toxicity. Giant unilamellar vesicles (GUVs) mimic the phospholipid bilayer of the cell membrane and are also useful drug delivery vehicles. Controlled increase of the permeability of GUVs is a delicate balance between sufficient perturbation for the delivery of the GUV contents and damage to the vesicles. Here we show that photoacoustic waves can promote the release of FITC-dextran or GFP from GUVs without damage. Real-time interferometric imaging offers the first movies of photoacoustic wave propagation and interaction with GUVs. The photoacoustic waves are seen as mostly compressive half-cycle pulses with peak pressures of?~?1 MPa and spatial extent FWHM?~?36 µm. At a repetition rate of 10 Hz, they enable the release of 25% of the FITC-dextran content of GUVs in 15 min. Such photoacoustic waves may enable non-invasive targeted release of GUVs and cell transfection over large volumes of tissues in just a few minutes.

SUBMITTER: Pereira DA

PROVIDER: S-EPMC7854711 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature

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Imaging of photoacoustic-mediated permeabilization of giant unilamellar vesicles (GUVs).

Pereira Diogo A DA Silva Alexandre D AD Martins Patricia A T PAT Piedade Ana P AP Martynowych Dmitro D Veysset David D Moreno Maria João MJ Serpa Carlos C Nelson Keith A KA Arnaut Luis G LG

Scientific reports 20210202 1

Target delivery of large foreign materials to cells requires transient permeabilization of the cell membrane without toxicity. Giant unilamellar vesicles (GUVs) mimic the phospholipid bilayer of the cell membrane and are also useful drug delivery vehicles. Controlled increase of the permeability of GUVs is a delicate balance between sufficient perturbation for the delivery of the GUV contents and damage to the vesicles. Here we show that photoacoustic waves can promote the release of FITC-dextra ...[more]

PMID: 33531539

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Project description:We developed a new (to our knowledge) protocol to generate giant unilamellar vesicles (GUVs) composed of mixtures of single lipopolysaccharide (LPS) species and Escherichia coli polar lipid extracts. Four different LPSs that differed in the size of the polar headgroup (i.e., LPS smooth > LPS-Ra > LPS-Rc > LPS-Rd) were selected to generate GUVs composed of different LPS/E. coli polar lipid mixtures. Our procedure consists of two main steps: 1), generation and purification of oligolamellar liposomes containing LPSs; and 2), electroformation of GUVs using the LPS-containing oligolamellar vesicles at physiological salt and pH conditions. Analysis of LPS incorporation into the membrane models (both oligolamellar vesicles and GUVs) shows that the final concentration of LPS is lower than that expected from the initial E. coli lipids/LPS mixture. In particular, our protocol allows incorporation of no more than 15 mol % for LPS-smooth and LPS-Ra, and up to 25 mol % for LPS-Rc and LPS-Rd (with respect to total lipids). We used the GUVs to evaluate the impact of different LPS species on the lateral structure of the host membrane (i.e., E. coli polar lipid extract). Rhodamine-DPPE-labeled GUVs show the presence of elongated micrometer-sized lipid domains for GUVs containing either LPS-Rc or LPS-Rd above 10 mol %. Laurdan GP images confirm this finding and show that this particular lateral scenario corresponds to the coexistence of fluid disordered and gel (LPS-enriched)-like micron-sized domains, in similarity to what is observed when LPS is replaced with lipid A. For LPSs containing the more bulky polar headgroup (i.e., LPS-smooth and LPS-Ra), an absence of micrometer-sized domains is observed for all LPS concentrations explored in the GUVs (up to ?15 mol %). However, fluorescence correlation spectroscopy (using fluorescently labeled LPS) and Laurdan GP experiments in these microscopically homogeneous membranes suggests the presence of LPS clusters with dimensions below our microscope's resolution (?380 nm radial). Our results indicate that LPSs can cluster into gel-like domains in these bacterial model membranes, and that the size of these domains depends on the chemical structure and concentration of the LPSs.

Dataset Information

Imaging of photoacoustic-mediated permeabilization of giant unilamellar vesicles (GUVs).

Publications

Imaging of photoacoustic-mediated permeabilization of giant unilamellar vesicles (GUVs).

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