Uptake, efflux, and mass transfer coefficient of fluorescent PAMAM dendrimers into pancreatic cancer cells
| Autor: | Kirk J. Czymmek, Eric Wickstrom, Norman J. Wagner, Armin W. Opitz |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
Dendrimers
Mass transfer coefficient Time Factors Macromolecular Substances Biophysics 02 engineering and technology Models Biological Biochemistry Article law.invention Efflux 03 medical and health sciences chemistry.chemical_compound Drug Delivery Systems Confocal microscopy law Dendrimer Cell Line Tumor Fluorescence microscope Polyamines Humans Fluorescein 030304 developmental biology Alexa Fluor Fluorescent Dyes 0303 health sciences PAMAM dendrimer Microscopy Confocal Models Statistical Dose-Response Relationship Drug Chemistry Cell Biology 021001 nanoscience & nanotechnology Fluoresceins Fluorescence Pancreatic Neoplasms Drug delivery Nanoparticles 0210 nano-technology Internalization |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Biomembranes. 1828(2):294-301 |
| ISSN: | 0005-2736 |
| DOI: | 10.1016/j.bbamem.2012.09.016 |
| Popis: | Targeted delivery of imaging agents to cells can be optimized with the understanding of uptake and efflux rates. Cellular uptake of macromolecules is studied frequently with fluorescent probes. We hypothesized that the internalization and efflux of fluorescently labeled macromolecules into and out of mammalian cells could be quantified by confocal microscopy to determine the rate of uptake and efflux, from which the mass transfer coefficient is calculated. The cellular influx and efflux of a third generation poly(amido amine) (PAMAM) dendrimer labeled with an Alexa Fluor 555 dye was measured in Capan-1 pancreatic cancer cells using confocal fluorescence microscopy. The Capan-1 cells were also labeled with 5-chloromethylfluorescein diacetate (CMFDA) green cell tracker dye to delineate cellular boundaries. A dilution curve of the fluorescently labeled PAMAM dendrimer enabled quantification of the concentration of dendrimer in the cell. A simple mass transfer model described the uptake and efflux behavior of the PAMAM dendrimer. The effective mass transfer coefficient was found to be 0.054±0.043μm/min, which corresponds to a rate constant of 0.035±0.023min−1 for uptake of the PAMAM dendrimer into the Capan-1 cells. The effective mass transfer coefficient was shown to predict the efflux behavior of the PAMAM dendrimer from the cell if the fraction of labeled dendrimer undergoing non-specific binding is accounted for. This work introduces a novel method to quantify the mass transfer behavior of fluorescently labeled macromolecules into mammalian cells. |
| Databáze: | OpenAIRE |
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