In vitro and in vivo platelet targeting by cyclic RGD-modified liposomes

J Biomed Mater Res A. 2010 Jun 1;93(3):1004-15. doi: 10.1002/jbm.a.32549.

Abstract

Cell-selective delivery using ligand-decorated nanoparticles is a promising modality for treating cancer and vascular diseases. We are developing liposome nanoparticles surface-modified by RGD peptide ligands having targeting specificity to integrin GPIIb-IIIa. This integrin is upregulated and stimulated into a ligand-binding conformation on the surface activated platelets. Activated-platelet adhesion and aggregation are primary events in atherosclerosois, thrombosis, and restenosis. Hence, platelet-targeted nanoparticles hold the promise of vascular site-selective delivery of drugs and imaging probes. Here, we report in vitro and ex vivo microscopy studies of platelet-targeting by liposomes surface-modified with a cyclic RGD peptide. The peptide-modified liposomes were labeled either with a lipophilic fluorophore or with lipid-tethered Nanogold(R). For in vitro tests, coverslip-adhered activated human platelets were incubated with probe-labeled liposomes, followed by analysis with fluorescence microscopy, phase contrast microscopy, and scanning electron microscopy (SEM). For in vivo tests, the liposomes were introduced within a catheter-injured carotid artery restenosis model in rats and post-euthanasia, the artery was imaged ex vivo by fluorescence microscopy and SEM. All microscopy results showed successful platelet-targeting by the peptide-modified liposomes. The in vitro SEM results also enabled visualization of nanoscopic liposomes attached to activated platelets. The results validate our nanoparticle design for site-selective vascular delivery.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Blood Platelets / cytology
  • Blood Platelets / drug effects*
  • Blood Platelets / metabolism*
  • Blood Platelets / ultrastructure
  • Carotid Arteries / drug effects
  • Carotid Arteries / pathology
  • Disease Models, Animal
  • Drug Delivery Systems / methods*
  • Flow Cytometry
  • Humans
  • Liposomes / pharmacology*
  • Microscopy, Fluorescence
  • Nanostructures / chemistry
  • Peptides, Cyclic / pharmacology*
  • Rats

Substances

  • Liposomes
  • Peptides, Cyclic
  • cyclic arginine-glycine-aspartic acid peptide