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Lipid-Coated, pH-Sensitive Magnesium Phosphate Particles for Intracellular Protein Delivery

  • Yunzhou Fang
  • Mallika Vadlamudi
  • Yingbo Huang
  • Xin GuoEmail author
Research Paper
  • 65 Downloads

Abstract

Purpose

To develop cationic lipid-coated magnesium phosphate nanoparticles (LPP) for intracellular catalase (CAT) delivery.

Methods

Magnesium phosphate nanoparticles (MgP NP) were prepared by micro-emulsion precipitation and mixed with catalase-loaded cationic liposomes (DOTAP/cholesterol) to yield LPP formulation of catalase (LPP-CAT). The size and ζ-potential of LPP-CAT were measured by dynamic light scattering. The pH-sensitivity of LPP-CAT was determined by monitoring their degradation of hydrogen peroxide (H2O2) and their morphologies under transmission electron microscopy (TEM) at pH 7.4 and 5.5. The ability of LPP-CAT to protect MCF-7 cells against hydrogen peroxide was measured by MTS assay. ROS levels in EA.hy926 cells were measured after treatment with LPP-CAT.

Results

LPP-CAT were successfully prepared and carried an average diameter of <300 nm and ζ -potential of about +40 mV. At pH 5.5, LPP-CAT degraded H2O2 almost 4-fold as fast as pH 7.4 and displayed drastic morphological changes of an osmotic explosion. LPP-CAT protected MCF-7 cells from lethal level of exogenous H2O2 and significantly lowered the ROS levels in EA.hy926 cells. A lipid with a pH-sensitive conformational switch (flipid) further enhanced the protein delivery of LPP-CAT.

Conclusion

LPP represents a promising nano-system for intracellular protein delivery.

KEY WORDS

intracellular delivery liposome magnesium phosphate nanoparticles protein delivery triggered release 

Abbreviations

CAT

Catalase

HBS

HEPES buffered saline (10 mM HEPES, 150 mM NaCl, pH 7.4)

L – CAT

Cationic liposomes loaded with catalase

LF – CAT

L – CAT containing flipid in the lipid membrane

LPP

Lipid-coated magnesium phosphate nanoparticles

LPP – CAT

Lipid-coated magnesium phosphate nanoparticles loaded with catalase

LPPF – CAT

LPP – CAT containing flipid in the lipid membrane

MgP NP

Magnesium phosphate nanoparticles

ROS

Reactive oxygen species

Flipid (F)

Bisdodecyl-4-hydroxy-trans-5-morpholinocyclohexane-trans-1,2-dicarboxylate, a lipidic amphiphile containing a pH-sensitive conformational switch

TEM

Transmission electron microscopy

Notes

Acknowledgments and Disclosures

This work was supported by National Institute of Health (GM107823). We thank the technical consultations from Professor Leaf Huang and associates at University of North Carolina, Chapel Hill. We thank Electron Imaging Facility at University of California, Davis for access to TEM. We thank Professor Xiaoling Li at University of the Pacific for access to UV spectrometer for kinetic analyses of catalase activities.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Pharmaceutics and Medicinal Chemistry, Thomas J. Long School of Pharmacy and Health SciencesUniversity of the PacificStocktonUSA

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