Brownian diffusion and surface kinetics of liposome and viral particle uptake by human lung cancer cells in-vitro.

ABSTRACT

In this study, the simultaneous roles of transport, diffusion, and surface kinetic uptake of liposome (Lip-FD), adenoviral (AD-Cy2), and liposome-adenoviral complex (lip-FD-Ad) particles by a non-small cell human lung cancer (A549) were examined through a coupling of in vitro experimental and mathematical modeling techniques. Experimentally, quantitative fluorescence spectroscopy was used to monitor time dependent particle uptake rates including low temperature (5 degrees C) conditions where endocytosis could be inhibited. Mathematically, analytic solutions to the Brownian particle diffusion equation with Langmuir type boundary conditions for the adsoprtion, desorption, and endocytosis process, were obtained for both unsteady and steady-state (no endocytosis) conditions. By direct comparisons of experimental data to model solutions, the adsorption constants, desorption constants, and number of cell surface receptor sites were determined for all particle types considered. It was found that the particle adsorption and desorption constants were of the same order of magnitude compared to earlier studies (Singh, M., T. Ghose, G. Faulkner, and M. Mezei. Cancer Res. 493976-3984, 1990.) using different cell lines, particle types, and methodologies. Also in agreement with previous studies using differing cell lines and methodologies (Miller, C. R., B. Boundurant, S. D. McLeon, K. A. McGovern, and D. F. O'Brien. Biochemistry 3712875-12883, 1998; Perry, D. G., and W. J. Martin II. J. Immunol. Methods 181269-285, 1995; Muller, W. J., K. Zen, A. B. Fisher, and H. Shuman. Am. J. Physiol. L11-L19, 1995), the number of cell surface receptor sites was predicted to be several orders of magnitude higher for liposome and liposome-viral complex than for viral particles alone, suggesting a nonspecific or nonrestrictive binding pattern for liposomes and liposome complexes and a specific or restrictive binding pattern for viral particles. The surface kinetic constants obtained here for the A549 cells may be useful in physiological modeling or pharmacokinetic applications of chemical or genetic carrying particles in the treatment of lung cancer and other lung diseases. Furthermore, the methodologies given here are straightforward and can be applied to other particle-cell uptake systems.