Uptake and Cytotoxicity of Docetaxel-Loaded Hyaluronic Acid-Grafted Oily Core Nanocapsules in MDA-MB 231 Cancer Cells
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It is hypothesized that docetaxel (Doc)-loaded hyaluronic acid (HA)-polyethylene glycol/poly(ε-caprolactone)-grafted oily core nanocapsules (NCs) can enhance the drug cytotoxicity and uptake in CD44 expressing breast cancer (BC) cells (MDA-MB 231).
NCs were prepared, optimized and characterized by dynamic light scattering, transmission electron microscopy (TEM), and powder X-ray diffraction (PXRD). In vitro cytotoxicity tests [MTS, level of reactive oxygen species (ROS) and level of reduced glutathione (GSH)] were performed in BC cells. The contribution of CD44 to the NCs cellular uptake was elucidated using an anti CD44 antibody blockage and a CD44 negative NIH3T3 cell line.
The optimum formulation of Doc-loaded HA oily core NCs had respective mean diameter, polydispersity, and drug encapsulation efficiency of 224.18 nm, 0.32, and 60.38%. The NCs appeared spherical with low drug crystallinity, while the drug release data fitted to first order equation. Compared to that of ungrafted NCs, the cytotoxicity of Doc-loaded HA-grafted NCs was significantly enhanced (p<0.05). A decrease of the intracellular level of ROS was reversely correlated with that of GSH. Interestingly, the cellular internalization of HA-grafted NCs mediated CD44 was dramatically enhanced (3 to 4-fold) with respect to the absence of specific biomarker or targeting ligand.
The use of HA-grafted NCs enhanced the selective drug payload, cytotoxicity and uptake in MDA-MB 231 cells. Therefore, it could be a promising template for safe and effective delivery of Doc and similar chemotherapeutic agents in cancer cells.
KEY WORDSbreast cancer cancer chemotherapy CD44 receptor cytotoxicity hyaluronic acid nanocapsules particle cellular uptake
ACKNOWLEDGMENTS AND DISCLOSURES
The work was supported by UMKC School of Pharmacy Dean internal bridge/seed funds. The authors acknowledge Gattefossé Corporation (Saint-Priest, France) for providing a gift sample of Labrafac CC oil. We gratefully acknowledge Mr. Jack Liu (Zhenjiang Dong Yuan Biotech Co., Ltd., Jiangsu, China) for providing samples of hyaluronic acid. We also thank Richard Hastings at Kansas University for his assistance with the flow cytometry analysis. We are grateful to Dr. Jeffrey L. Price for providing assistance with the confocal laser scanning microscopy at the University of Missouri, Kansas City; School of Biological Sciences. The authors appreciate the helpful discussions on ROS and GSH assay with Dr. Miezan JM Ezoulin (Post-doctoral research associate at the University of Missouri Kansas City; School of Pharmacy. The assistance of Jianing Meng (Graduate student at the University of Missouri Kansas City School of Pharmacy) for the figure graphic design is appreciated. The authors thank Barbara Fegley for her assistance with the Electron microscopy at University of Kansas Medical Center, Kansas City, Kansas.
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