ABSTRACT
The surface of polymeric nanocapsules used as ultrasound contrast agents (UCAs) was modified with PEGylated phospholipids in order to escape recognition and clearance by the mononuclear phagocyte system and achieve passive tumor targeting. Nanocapsules consisted of a shell of poly(lactide-co-glycolide) (PLGA) encapsulating a liquid core of perfluorooctyl bromide (PFOB). They were decorated with poly(ethylene glycol-2000)-grafted distearoylphosphatidylethanolamine (DSPE-PEG) incorporated in the organic phase before the solvent emulsification-evaporation process. The influence of DSPE-PEG concentration on nanocapsule size, surface charge, morphology, hydrophobicity and complement activation was evaluated. Zeta potential measurements, Hydrophobic interaction chromatography and complement activation provide evidence of DSPE-PEG presence at nanocapsule surface. Electronic microscopy reveals that the core/shell structure is preserved up to 2.64 mg of DSPE-PEG for 100 mg PLGA. In vivo ultrasound imaging was performed in mice bearing xenograft tumor with MIA PaCa-2 cells, either after an intra-tumoral or intravenous injection of nanocapsules. Tumor was observed only after the intra-tumoral injection. Despite the absence of echogenic signal in the tumor after intravenous injection of nanocapsules, histological analysis reveals their accumulation within the tumor tissue demonstrating that tissue distribution is not the unique property required for ultrasound contrast agents to be efficient.
Subject(s)
Contrast Media , Fluorocarbons , Nanocapsules , Polyethylene Glycols/chemistry , Ultrasonics , Animals , Chromatography , Complement Activation/immunology , Complement C3/immunology , Electrophoresis, Gel, Two-Dimensional , Humans , Hydrocarbons, Brominated , Hydrophobic and Hydrophilic Interactions , Mice , Nanocapsules/ultrastructure , Pancreatic Neoplasms/diagnostic imaging , Pancreatic Neoplasms/pathology , Particle Size , Phosphatidylethanolamines/chemistry , Surface Properties , UltrasonographyABSTRACT
Perfluorocarbons (PFCs) are fluorinated compounds that have been used for many years in clinics mainly as gas/oxygen carriers and for liquid ventilation. Besides this main application, PFCs have also been tested as contrast agents for ultrasonography and magnetic resonance imaging since the end of the 1970s. However, most of the PFCs applied as contrast agents for imaging were gaseous. This class of PFCs has been recently substituted by liquid PFCs as ultrasound contrast agents. Additionally, liquid PFCs are being tested as contrast agents for (19)F magnetic resonance imaging (MRI), to yield dual contrast agents for both ultrasonography and (19)F MRI. This review focuses on the development and applications of the different contrast agents containing liquid perfluorocarbons for ultrasonography and/or MRI: large and small size emulsions (i.e. nanoemulsions) and nanocapsules.
Subject(s)
Contrast Media/chemistry , Drug Carriers , Fluorine , Fluorocarbons , Magnetic Resonance Imaging/methods , Ultrasonography/methods , Animals , Drug Carriers/chemistry , Drug Delivery Systems , Fluorocarbons/chemistry , Humans , Models, Biological , Molecular Structure , Nanotechnology/methodsABSTRACT
We present here an easy method to modify the surface chemistry of polymeric microcapsules of perfluorooctyl bromide used as ultrasound contrast agents (UCAs). Capsules were obtained by a solvent emulsification-evaporation process with phospholipids incorporated in the organic phase before emulsification. Several phospholipids were reviewed: fluorescent, pegylated and biotinylated phospholipids. The influence of phospholipid concentration on microcapsule size and morphology was evaluated. Only a fraction of the phospholipids is associated to microcapsules, the rest being dissolved with the surfactant in the aqueous phase. Microscopy shows that phospholipids are present within the shell and that the core/shell structure is preserved up to 0.5 mg fluorescent phospholipids, up to about 0.25 mg pegylated phospholipids or biotinylated phospholipids (for 100 mg of polymer, poly(lactide-co-glycolide) (PLGA)). HPLC allows quantifying phospholipids associated to capsules: they correspond to 10% of pegylated phospholipids introduced in the organic phase. The presence of pegylated lipids at the surface of capsules was confirmed by X-ray photon electron spectroscopy (XPS). The pegylation did not modify the echographic signal arising from capsules. Finally biotinylated microcapsules incubated with neutravidin tend to aggregate, which confirms the presence of biotin at the surface. These results are encouraging and future work will consist of nanocapsule surface modification for molecular imaging.