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Coupling in vitro cell culture with synchrotron SAXS to understand the bio-interaction of lipid-based liquid crystalline nanoparticles with vascular endothelial cells

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Abstract

Nonlamellar lipid-based liquid crystalline (LLC) nanoparticles possessing different internal nanostructures, specifically the 3D-ordered cubosomes (V2 phase) and the 2D-ordered hexosomes (H2 phase), are of increasing interest as drug delivery systems. To facilitate their development, it is important that we understand their interactions with healthy human umbilical vein endothelial cells (HUVECs). To this end, a 3D cells-in-a-tube model that recapitulates the basic morphology (i.e. tubular lumen) and in vivo microenvironment (i.e. physiological shear stress) of blood vessels was employed as a biomimetic testing platform, and the bio-nanoparticle interactions were compared with that of the conventional 2D planar cell culture. Confocal microscopy imaging revealed internalisation of the nanoparticles into HUVECs within 2 h and that the nanoparticle-cell interactions of cubosomes and hexosomes were not significantly different from one another. Low fluid shear stress conditions (i.e. venous simulation at 0.8 dynes/cm2) were shown to impose subtle effects on the degree of nanoparticle-cell interactions as compared with the static 2D culture. The unexpected similarity of cellular interactions between cubosomes and hexosomes was clarified via a real-time phase behaviour analysis using the synchrotron-based small-angle X-ray scattering (SAXS) technique. When the nanoparticles came into contact with HUVECs under circulating conditions, the cubosomes gradually evolved into hexosomes (within 16 min). In contrast, the hexosomes retained their original internal structure with minimal changes to the lattice parameters. This study highlights the need to couple cellular studies with high-resolution analytics such as time-resolved SAXS analysis to ensure that particle structures are verified in situ, enabling accurate interpretation of the dynamics of cellular interactions and potential bio-induced changes of particles intended for biomedical applications.

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Acknowledgements

Mr. Cameron J. Nowell assisted the confocal microscopy imaging and analysis at the imaging, FACS, and Analysis Core, Monash University. The cryo-TEM images were taken at the Bio21 Institute with the assistance of Ms. Xiaohan Sun (Monash PhD candidate). The SAXS experiments were undertaken on the SAXS/WAXS beamline at the ANSTO Australian Synchrotron, Victoria.

Funding

This project is funded by the Australian Research Council (ARC) Centre of Excellence in Convergent Bio-Nano Science and Technology. B.J.B. was the recipient of an Australian Research Council Future Fellowship. Y.Y.L. was supported by a Monash University Jubilee Honours Scholarship.

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Authors and Affiliations

  1. Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University (Parkville Campus), 381 Royal Parade, Parkville, VIC, 3052, Australia

    Yuen Yi Lam, Angel Tan & Ben J. Boyd

  2. ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, Monash Institute of Pharmaceutical Sciences, Monash University (Parkville Campus), Parkville, VIC, 3052, Australia

    Yuen Yi Lam, Angel Tan & Ben J. Boyd

  3. SAXS/WAXS beamline, Australian Synchrotron, ANSTO, Clayton, VIC, 3168, Australia

    Adrian Hawley

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  1. Yuen Yi Lam
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Correspondence to Angel Tan or Ben J. Boyd.

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Lam, Y.Y., Hawley, A., Tan, A. et al. Coupling in vitro cell culture with synchrotron SAXS to understand the bio-interaction of lipid-based liquid crystalline nanoparticles with vascular endothelial cells. Drug Deliv. and Transl. Res. 10, 610–620 (2020). https://doi.org/10.1007/s13346-020-00718-3

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