Mechanism of cellular uptake of highly fluorescent conjugated polymer nanoparticles

TitleMechanism of cellular uptake of highly fluorescent conjugated polymer nanoparticles
Publication TypeJournal Article
Year of Publication2010
AuthorsFernando, L. P., Kandel P. K., Yu J., McNeill J., Ackroyd P. C., and Christensen K. A.
JournalBiomacromolecules
Volume11
Pagination2675-82
Date PublishedOct 11
ISBN Number1526-4602 (Electronic)<br/>1525-7797 (Linking)
Accession Number20863132
Keywords*Fluorescence, Animals, Cell Culture Techniques, Cell Line, Flow cytometry, Fluorenes/chemistry/*metabolism, Macrophages/*metabolism, Mice, Microscopy, Atomic Force, Microscopy, Fluorescence, Molecular Imaging/*methods, Nanoparticles/*chemistry, Polymers/chemistry/*metabolism
Abstract

Conjugated polymer nanoparticles are formed by precipitation of highly fluorescent conjugated polymers to form small nanoparticles with extremely bright fluorescence. We characterized cellular uptake and cytotoxicity of 18 +/- 5 nm PFBT conjugated polymer nanoparticles in J774A.1 cells. Significant nanoparticle uptake was observed, indicating efficient nanoparticle entry into cells, even for short (1 h) incubations. The high fluorescence of these nanoparticles allows extremely low loading concentrations; PFBT nanoparticle fluorescence in cells could be detected with loading concentrations of 155 pM (270 ppb). Cellular uptake slows at low temperature, consistent with endocytic entry. Nanoparticles colocalize with Texas Red dextran and are trafficked to lysosomes, as demonstrated by the location of nanoparticle fluorescence in perinuclear organelles that also stain with an anti-LAMP-1 antibody. Inhibition of uptake by phosphoinositide 3-kinase inhibitors implicates macropinocytosis as the operative endocytic mechanism. No significant cytotoxic or inflammatory effects could be observed, making PFBT nanoparticles attractive probes for live cell imaging.

Short TitleMechanism of Cellular Uptake of Highly Fluorescent Conjugated Polymer Nanoparticles