Cytotoxicity of doxorubicin bound to poly(butyl cyanoacrylate) nanoparticles in rat glioma cell lines using different assays

The cytotoxicity of doxorubicin bound to poly(butyl cyanoacrylate) nanoparticles (Dox-PBCA-NP) was investigated in the rat glioma cell lines GS-9L, F-98 and RG-2. MTT and LDH assays were used as cytotoxic assays. In general, the cytotoxicity of nanoparticle-bound doxorubicin (Dox) was enhanced compared to the free drug in solution. However, responses of the cell lines towards the drug effects were different. In the case of free Dox in solution, this difference correlated with different intracellular concentrations of Dox, which in turn, depended on the level of P-glycoprotein (P-gp) expression in these cell lines. Accordingly, the 9L gliosarcoma (GS-9L) cells, which appeared to be most resistant towards Dox, were characterized by the highest P-gp expression. Additionally, the influence of surfactants on the cytotoxic effect was investigated at different Dox concentrations. It was shown that the presence of polysorbate 80 (Tween® 80) in the nanoparticle formulation significantly enhanced the cytotoxicity, whereas poloxamer 188 (Pluronic® F68) and poloxamine 908 (Tetronic® 908) had a negligible influence.

[1]  M. Chevallier,et al.  Increase of doxorubicin sensitivity by doxorubicin-loading into nanoparticles for hepatocellular carcinoma cells in vitro and in vivo. , 2005, Journal of hepatology.

[2]  B. Sangro.  Refined tools for the treatment of hepatocellular carcinoma. , 2005, Journal of hepatology.

[3]  Andreas Zimmer,et al.  A practical note on the use of cytotoxicity assays. , 2005, International journal of pharmaceutics.

[4]  V. P. Collins,et al.  Uptake of Adriamycin in tumour and surrounding brain tissue in patients with malignant gliomas , 1990, Acta Neurochirurgica.

[5]  D. Begley.  Delivery of therapeutic agents to the central nervous system: the problems and the possibilities. , 2004, Pharmacology & therapeutics.

[6]  K. Geiger,et al.  Chemotherapy of glioblastoma in rats using doxorubicin‐loaded nanoparticles , 2004, International journal of cancer.

[7]  D. Begley,et al.  Direct Evidence That Polysorbate-80-Coated Poly(Butylcyanoacrylate) Nanoparticles Deliver Drugs to the CNS via Specific Mechanisms Requiring Prior Binding of Drug to the Nanoparticles , 2003, Pharmaceutical Research.

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

[9]  R. Borchardt,et al.  The Use of Surfactants to Enhance the Permeability of Peptides Through Caco-2 Cells by Inhibition of an Apically Polarized Efflux System , 1996, Pharmaceutical Research.

[10]  Rolf F. Barth,et al.  Rat brain tumor models in experimental neuro-oncology:The 9L, C6, T9, F98, RG2 (D74), RT-2 and CNS-1 Gliomas , 2004, Journal of Neuro-Oncology.

[11]  T. Yamashima,et al.  Uptake of drugs and expression of P-glycoprotein in the rat 9L glioma , 2004, Experimental Brain Research.

[12]  W. Pardridge,et al.  P-glycoprotein and caveolin-1alpha in endothelium and astrocytes of primate brain. , 2003, Neuroreport.

[13]  K. Harada,et al.  Repetitive doxorubicin treatment of glioblastoma enhances the PGP expression--a special role for endothelial cells. , 2003, Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie.

[14]  R. Bendayan,et al.  Drug transporters in the central nervous system: brain barriers and brain parenchyma considerations. , 2001, Pharmacological reviews.

[15]  R. Bendayan,et al.  Functional expression of P-glycoprotein in rat brain microglia. , 2001, The Journal of pharmacology and experimental therapeutics.

[16]  B. McEwen,et al.  Neurotrophic and neuroprotective actions of estrogens and their therapeutic implications. , 2001, Annual review of pharmacology and toxicology.

[17]  P Couvreur,et al.  Reversion of multidrug resistance by co-encapsulation of doxorubicin and cyclosporin A in polyalkylcyanoacrylate nanoparticles. , 2000, Biomaterials.

[18]  J. Dichgans,et al.  Glioblastoma multiforme: Mechanisms of resistance to chemotherapy , 1999 .

[19]  W. Berger,et al.  Expression of the multidrug resistance‐associated protein (MRP) and chemoresistance of human non‐small‐cell lung cancer cells , 1997 .

[20]  P. Couvreur,et al.  On the mechanism of action of doxorubicin encapsulation in nanospheres for the reversal of multidrug resistance , 1996, Cancer Chemotherapy and Pharmacology.

[21]  Susan M. Chang,et al.  Intratumoral chemotherapy. Commentaries , 1995 .

[22]  A. Olivi,et al.  Intratumoral chemotherapy. , 1995, Neurosurgery.

[23]  K. Langer,et al.  Characterisation of polybutylcyanoarylate nanoparticles: I. Quantification of PCBA polymer and dextrans , 1994 .

[24]  W. Sawyer,et al.  Reversal of multidrug resistance by surfactants. , 1992, British journal of cancer.

[25]  J. Bulte,et al.  Monoclonal antibody JSB‐1 detects a highly conserved epitope on the P‐glycoprotein associated with multi‐drug‐resistance , 1988, International journal of cancer.

[26]  J. Kreuter.  Evaluation of nanoparticles as drug-delivery systems. II: Comparison of the body distribution of nanoparticles with the body distribution of microspheres (diameter greater than 1 micron), liposomes, and emulsions. , 1983, Pharmaceutica acta Helvetiae.

[27]  P. Couvreur,et al.  Polycyanoacrylate nanocapsules as potential lysosomotropic carriers: preparation, morphological and sorptive properties , 1979, The Journal of pharmacy and pharmacology.

[28]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.