Research ArticlesMechanistic study of the uptake of wheat germ agglutinin-conjugated PLGA nanoparticles by A549 cells
Section snippets
INTRODUCTION
Bioadhesive delivery systems are formulated to enhance drug bioavailability by providing more intimate epithelial contact and prolonged residence time at the site of absorption.1 Early prototypes that attached predominantly to the mucous layer overlaying epithelial cells (mucoadhesive systems) often failed to live up to expectations because of their interactions with soluble mucins and the rapid mucus turnover in vivo.2 This problem might be overcome by developing drug delivery systems that
Materials
Resomer® RG 502H (lactide:glycolide = 50:50, acid number of 10.7 mg KOH/g) was a gift of Boehringer Ingelheim, Germany. Acetone, FITC-labeled WGA (fWGA, 2.5 mol FITC per mol WGA), FITC-labeled BSA (fBSA, 12 mol FITC per mol BSA), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDAC), N-hydroxy-succinimide (NHS), phosphotungstic acid (PTA), polyvinyl alcohol (PVA, MW 30,000–70,000), glycine, propodium iodide (PI), sodium dodecyl sulfate (SDS), N-(2-hydroxyethyl)
RESULTS
The two-step EDAC/NHS method23 enables the conjugation of protein amino groups to the PLGA carboxyl groups in an aqueous medium through an amide bond formation. In this study, the conjugation efficiency was 22.87 ± 1.33 and 24.25 ± 1.5 μg/mg for the fWGA-PLGA nanoparticles and fBSA-PLGA nanoparticles, respectively. Physically adsorbed fWGA amounted to 2.84 ± 1.16 μg/mg, equivalent to 12.4% of the total surface WGA on the nanoparticles.
The grafting of surface proteins modified the mean size,
DISCUSSION
fWGA and fBSA were successfully grafted onto the PLGA nanoparticles by the two-step carbodiimide method. The conjugation efficiencies obtained for the two proteins were similar, but they were significantly higher than those reported for PLGA microspheres of 4-μm diameter.17 The higher conjugation efficiency could be attributed to the larger specific surface area available for grafting in the nanoparticles, together with the shorter activation time (0.5 versus 3.5 h) employed in this study. We
CONCLUSION
The grafting of wheat germ agglutinin (WGA), a plant lectin with established cytoadhesive and cytoinvasive properties, onto the surface of pre-formed PLGA nanoparticles at a density of 22.87 mg WGA/mg nanoparticle by a two-step carbodiimide method did not block receptor recognition and subsequent internalization of the lectin. The grafted WGA enhanced by five- to eightfold the uptake of the nanoparticles by A549 cells when compared to the cellular uptake of PLGA nanoparticles similarly grafted
Acknowledgements
This study was supported by a National University of Singapore grant (R148-000-023-112). Yun Mo is grateful to the National University of Singapore for financial support of her graduate studies.
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