Self-Assembled Biodegradable Nanoparticles Developed by Direct Dialysis for the Delivery of Paclitaxel

PurposeThe main objective of this study was to obtain self-assembled biodegradable nanoparticles by a direct dialysis method for the delivery of anticancer drug. The in vitro cellular particle uptake and cytotoxicity to C6 glioma cell line were investigated.MethodsSelf-assembled anticancer drugs—paclitaxel-loaded poly(d,l-lactic-co-glycolic acid) (PLGA) and poly(l-lactic acid) (PLA) nanoparticles—were achieved by direct dialysis. The physical and chemical properties of nanoparticles were characterized by various state-of-the-art techniques. The encapsulation efficiency and in vitro release profile were measured by high-performance liquid chromatography. Particle cellular uptake was studied using confocal microscopy, microplate reader, and flow cytometry. In addition, the cytotoxicity of this drug delivery system was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay on C6 glioma cell line to predict the possible dose response of paclitaxel-loaded PLGA and PLA nanoparticles.ResultsPLGA and PLA nanoparticles with or without vitamin E tocopherol polyethylene glycol succinate (TPGS) as an additive were obtained, in which the sustained release of paclitaxel of more than 20 days was achieved. The coumarin6-loaded PLGA and PLA nanoparticles could penetrate the C6 glioma cell membrane and be internalized. The cytotoxicity of paclitaxel-loaded nanoparticles seemed to be higher than that of commercial Taxol® after 3 days incubation when paclitaxel concentrations were 10 and 20 μg/ml.ConclusionsDirect dialysis could be employed to achieve paclitaxel-loaded PLGA and PLA nanoparticles, which could be internalized by C6 glioma cells and enhance the cytotoxicity of paclitaxel because of its penetration to the cytoplasm and sustained release property.

[1]  Gordon L. Amidon,et al.  Gastrointestinal Uptake of Biodegradable Microparticles: Effect of Particle Size , 1996, Pharmaceutical Research.

[2]  L. Lim,et al.  Mechanistic study of the uptake of wheat germ agglutinin-conjugated PLGA nanoparticles by A549 cells. , 2004, Journal of pharmaceutical sciences.

[3]  K. Audus,et al.  Uptake of surfactant-coated poly(methyl methacrylate)-nanoparticles by bovine brain microvessel endothelial cell monolayers , 1994 .

[4]  S. Davis,et al.  PLGA nanoparticles prepared by nanoprecipitation: drug loading and release studies of a water soluble drug. , 1999, Journal of controlled release : official journal of the Controlled Release Society.

[5]  R. Luduena,et al.  Characterization of nuclear betaII-tubulin in tumor cells: a possible novel target for taxol. , 2002, Cell motility and the cytoskeleton.

[6]  M. Davies,et al.  The colloidal properties of surfactant-free biodegradable nanospheres from poly(β-malic acid-co-benzyl malate)s and poly(lactic acid-co-glycolide) , 1995 .

[7]  C. Lipschultz,et al.  Cytotoxic studies of paclitaxel (Taxol) in human tumour cell lines. , 1993, British Journal of Cancer.

[8]  W. Mellado,et al.  Taxol: mechanisms of action and resistance. , 1986, Annals of the New York Academy of Sciences.

[9]  S. Feng,et al.  Vitamin E TPGS used as emulsifier in the solvent evaporation/extraction technique for fabrication of polymeric nanospheres for controlled release of paclitaxel (Taxol). , 2002, Journal of controlled release : official journal of the Controlled Release Society.

[10]  M. Marsh,et al.  Endocytosis: what goes in and how? , 1992, Journal of cell science.

[11]  M. Schlame,et al.  HDL-holoparticle uptake by alveolar type II cells: effect of vitamin E status. , 2002, American journal of respiratory cell and molecular biology.

[12]  C. Regan,et al.  Transient heat shock in mid-G1-phase of the C6 glioma cell cycle impairs entry into S-phase. , 1991, Toxicology letters.

[13]  H. Lentzen,et al.  Phagocytotic activity of glial cells in culture. , 1982, Experimental cell research.

[14]  M. Jordan,et al.  Mechanism of mitotic block and inhibition of cell proliferation by taxol at low concentrations. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[15]  T. Kissel,et al.  Biodegradable nanoparticles for oral delivery of peptides: is there a role for polymers to affect mucosal uptake? , 2000, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[16]  S. Simões,et al.  Paclitaxel-loaded PLGA nanoparticles: preparation, physicochemical characterization and in vitro anti-tumoral activity. , 2002, Journal of controlled release : official journal of the Controlled Release Society.

[17]  Jayanth Panyam,et al.  Fluorescence and electron microscopy probes for cellular and tissue uptake of poly(D,L-lactide-co-glycolide) nanoparticles. , 2003, International journal of pharmaceutics.

[18]  Jian-Jiang Zhong,et al.  Plant cell culture for production of paclitaxel and other taxanes. , 2002, Journal of bioscience and bioengineering.

[19]  P. Couvreur,et al.  Nanoparticles as microcarriers for anticancer drugs , 1990 .

[20]  D. Aggarwal,et al.  Paclitaxel and its formulations. , 2002, International journal of pharmaceutics.

[21]  V. Labhasetwar,et al.  Characterization of nanoparticle uptake by endothelial cells. , 2002, International journal of pharmaceutics.

[22]  J. Manfredi,et al.  Taxol binds to cellular microtubules , 1982, The Journal of cell biology.

[23]  G. Griffiths,et al.  Phagocytosis: latex leads the way. , 2003, Current opinion in cell biology.

[24]  A. Lowman,et al.  Biodegradable nanoparticles for drug delivery and targeting , 2002 .

[25]  S. Feng,et al.  Methoxy poly(ethylene glycol)-poly(lactide) (MPEG-PLA) nanoparticles for controlled delivery of anticancer drugs. , 2004, Biomaterials.

[26]  B. Monsarrat,et al.  Taxol: pharmacology, metabolism and clinical implications. , 1993, Cancer surveys.

[27]  D. Ettinger,et al.  Taxol: a unique antineoplastic agent with significant activity in advanced ovarian epithelial neoplasms. , 1989 .

[28]  W. Zauner,et al.  In vitro uptake of polystyrene microspheres: effect of particle size, cell line and cell density. , 2001, Journal of controlled release : official journal of the Controlled Release Society.

[29]  E. Raymond,et al.  Effects of prolonged versus short-term exposure paclitaxel (Taxol) on human tumor colony-forming units. , 1997, Anti-cancer drugs.

[30]  E. Rowinsky,et al.  Taxol: A unique antineoplastic agent with significant activity in advanced ovarian epithelial neoplasms , 1990, Annals of internal medicine.

[31]  D. Bazile,et al.  Release of the fluorescent marker Prodan® from poly(D,L-lactic acid) nanoparticles coated with albumin or polyvinyl alcohol in model digestive fluids (USP XXII) , 1997 .

[32]  J. Kreuter,et al.  Significant Transport of Doxorubicin into the Brain with Polysorbate 80-Coated Nanoparticles , 1999, Pharmaceutical Research.

[33]  C Zimmer,et al.  MR imaging of phagocytosis in experimental gliomas. , 1995, Radiology.

[34]  J. Nah,et al.  Preparation of poly(DL‐lactide‐co‐glycolide) nanoparticles without surfactant , 2001 .

[35]  F. Liu,et al.  Cellular Uptake Study of Biodegradable Nanoparticles in Vascular Smooth Muscle Cells , 1998, Pharmaceutical Research.

[36]  S. Davis,et al.  The importance of gastrointestinal uptake of particles in the design of oral delivery systems , 1995 .

[37]  John Samuel,et al.  Analysis of Poly(D,L-Lactic-Co-Glycolic Acid) Nanosphere Uptake by Human Dendritic Cells and Macrophages In Vitro , 2002, Pharmaceutical Research.

[38]  K. Audus,et al.  Microparticulate uptake mechanisms of in‐vitro cell culture models of the respiratory epithelium , 2001, The Journal of pharmacy and pharmacology.

[39]  Y. M. Lee,et al.  Clonazepam release from core-shell type nanoparticles in vitro. , 1998, Journal of controlled release : official journal of the Controlled Release Society.

[40]  J. Nah,et al.  Surfactant‐free nanoparticles of poly(DL‐lactide‐co‐glycolide) prepared with poly(L‐lactide)/ poly(ethylene glycol) , 2003 .

[41]  D. Begley,et al.  Polysorbate‐80 coating enhances uptake of polybutylcyanoacrylate (PBCA)‐nanoparticles by human and bovine primary brain capillary endothelial cells , 2000, The European journal of neuroscience.

[42]  A. R. Kulkarni,et al.  Biodegradable polymeric nanoparticles as drug delivery devices. , 2001, Journal of controlled release : official journal of the Controlled Release Society.

[43]  H. Merkle,et al.  Transfer of Lipophilic Markers from PLGA and Polystyrene Nanoparticles to Caco-2 Monolayers Mimics Particle Uptake , 2002, Pharmaceutical Research.

[44]  Gordon L. Amidon,et al.  The Mechanism of Uptake of Biodegradable Microparticles in Caco-2 Cells Is Size Dependent , 1997, Pharmaceutical Research.

[45]  G. Hortobagyi,et al.  Phase II trial of taxol, an active drug in the treatment of metastatic breast cancer. , 1991, Journal of the National Cancer Institute.

[46]  S. Sahoo,et al.  Residual polyvinyl alcohol associated with poly (D,L-lactide-co-glycolide) nanoparticles affects their physical properties and cellular uptake. , 2002, Journal of controlled release : official journal of the Controlled Release Society.

[47]  S. Feng,et al.  Effects of emulsifiers on the controlled release of paclitaxel (Taxol) from nanospheres of biodegradable polymers. , 2001, Journal of controlled release : official journal of the Controlled Release Society.

[48]  D. W. Pack,et al.  Polymeric nanospheres fabricated with natural emulsifiers for clinical administration of an anticancer drug paclitaxel (Taxol , 2002 .

[49]  M. Ott,et al.  Paclitaxel shows cytotoxic activity in human hepatocellular carcinoma cell lines. , 1999, Cancer letters.

[50]  R. A. Jain,et al.  The manufacturing techniques of various drug loaded biodegradable poly(lactide-co-glycolide) (PLGA) devices. , 2000, Biomaterials.

[51]  D D Allen,et al.  Nanoparticle Technology for Drug Delivery Across the Blood-Brain Barrier , 2002, Drug development and industrial pharmacy.

[52]  Shu Chien,et al.  Chemotherapeutic engineering: Application and further development of chemical engineering principles for chemotherapy of cancer and other diseases , 2003 .