Cytotoxicity of Paclitaxel in Biodegradable Self-Assembled Core-Shell Poly(Lactide-Co-Glycolide Ethylene Oxide Fumarate) Nanoparticles
Biodegradable core-shell polymeric nanoparticles (NPs), with a hydrophobic core and hydrophilic shell, are developed for surfactant-free encapsulation and delivery of Paclitaxel to tumor cells.
Poly (lactide-co-glycolide fumarate) (PLGF) and Poly (lactide-fumarate) (PLAF) were synthesized by condensation polymerization of ultra-low molecular weight poly(l-lactide-co-glycolide) (ULMW PLGA) with fumaryl chloride (FuCl). Similarly, poly(lactide-co-ethylene oxide fumarate) (PLEOF) macromer was synthesized by reacting ultra-low molecular weight poly(l-lactide) (ULMW PLA) and PEG with FuCl. The blend PLGF/PLEOF and PLAF/PLEOF macromers were self-assembled into NPs by dialysis. The NPs were characterized with respect to particle size distribution, morphology, and loading efficiency. The physical state and miscibility of Paclitaxel in NPs were characterized by differential scanning calorimetry. Tumor cell uptake and cytotoxicity of Paclitaxel loaded NPs were measured by incubation with HCT116 human colon carcinoma cells. The distribution of NPs in vivo was assessed with ApcMin/+mouse using infrared imaging.
PLEOF macromer, due to its amphiphilic nature, acted as a surface active agent in the process of self-assembly which produced core-shell NPs with PLGF/PLAF and PLEOF macromers as the core and shell, respectively. The encapsulation efficiency ranged from 70 to 56% and it was independent of the macromer but decreased with increasing concentration of Paclitaxel. Most of the PLGF and PLAF NPs degraded in 15 and 28 days, respectively, which demonstrated that the release was dominated by hydrolytic degradation and erosion of the matrix. As the concentration of Paclitaxel was increased from 0 to 10, and 40 μg/ml, the viability of HCT116 cells incubated with free Paclitaxel decreased from 100 to 65 and 40%, respectively, while those encapsulated in PLGF/PLEOF NPs decreased from 93 to 54 and 28%.
Groups with Paclitaxel loaded NPs had higher cytotoxicity compared to Paclitaxel directly added to the media at the same concentration. NPs acted as reservoirs to protect the drug from epimerization and hydrolysis while providing a sustained dose of Paclitaxel with time. Infrared image of the ApcMin/+ mouse injected with NPs showed significantly higher concentration of NPs in the intestinal tissue.
Key wordsbiodegradable nanoparticle core-shell morphology self-assembly tumor drug delivery
This publication was made possible in part by NIH Grant No. P20 RR-016461 from the National Center for Research Resources and by the National Science Foundation/EPSCoR under Grant No. 2001 RII-EPS-0132573. This work was also supported by grants from the AO (Arbeitsgemeinschaft Fur Osteosynthesefragen) Foundation (AORF project 05-J95), and the Aircast Foundation. E. Jabbari thanks Dr. Frank Berger (Center for Colon Cancer Research) at the University of South Carolina for providing the ApcMin/+ mouse. E. Jabbari thanks Kelley Intehar (LI-COR Biosciences) for providing the IRDye 800RS Carboxylate dye and scanning the mice with the Odyssey Infrared Imaging System.
- 1.ACS. Cancer Facts & Figures. American Cancer Society, Atlanta, GA, Website: http://www.cancer.org/downloads/STT/CAFF2007PWSecured.pdf, (2007).
- 3.S. Gagandeep, P. M. Novikoff, M. Ott, and S. Gupta. Paclitaxel shows cytotoxic activity in human hepatocellular carcinoma cell lines. Cancer Res. 136:109 (1999).Google Scholar
- 10.M. Nahar, T. Dutta, S. Murugesan, A. Senthilkumar, A. Asthana, D. Abhay, D. Mishra, V. Rajkumar, M. Tare, S. Saraf, and N. Kumar. Functional polymeric nanoparticles: an efficient and promising tool for active delivery of bioactives. Critical Rev. Therap. Drug Carrier Sys. 23:259 (2006).Google Scholar
- 19.S. Unezaki, K. Maruyama, J.-I. Hosoda, I. Nagae, Y. Koyanagi, M. Nakata, O. Ishida, M. Iwatsuru, and S. Tsuchiya. Direct measurement of the extravasation of polyethyleneglycol-coated liposomes into solid tumor tissue by in vivo fluorescence microscopy. Int. J. Pharm. 144:11 (1996).CrossRefGoogle Scholar
- 33.E. Jababri, and X. He. Release characteristics of novel bioresorbable in-situ crosslinkable self assembled nanoparticles. CRS Abstract. CS:1 (2007).Google Scholar
- 34.E. Jabbari, and X. He. Synthesis and characterization of bioresorbable in situ crosslinkable ultra low molecular weight poly(lactide) macromer. J. Mater. Sci. Mater. Med. PMID:17597374 (2007).Google Scholar
- 37.E. Jabbari, and X. He. Synthesis and material properties of functionalized lactide oligomers as in situ crosslinkable scaffolds for tissue regeneration. Polym. Prepr. 47:353 (2006).Google Scholar
- 40.A. Sparreboom, J. van Asperen, U. Mayer, A. H. Schinkel, J. W. Smit, D. K. Meijer, P. Borst, W. J. Nooijen, J. H. Beijnen, and O. van Tellingen. Limited oral bioavailability and active epithelial excretion of paclitaxel (Taxol) caused by P-glycoprotein in the intestine. Proc. Natl. Acad. Sci. USA 94:2031 (1997).PubMedCrossRefGoogle Scholar
- 47.E. Jabbari, W. Xu, and X. He. Degradation characteristics of novel in-situ crosslinkable poly(lactide-co-glycolide-ethylene oxide-fumarate) copolymer networks. Trans. Soc. Biomaterials. 1:353 (2007).Google Scholar
- 50.M. Louis, D. Beck, D. Merkle, and G. Ciraolo. Particle size does not affect the rate of intracellular routing for ligands internalized by non-adsorptive pinocytosis. J. Electron Microsc. 46:337 (1997).Google Scholar
- 52.H. L. Wong, R. Bendayan, A. M. Rauth, H. Y. Xue, K. Babakhanian, and X. Y. Wu. A mechanistic study of enhanced doxorubicin uptake and retention in multidrug resistant breast cancer cells using a polymer-lipid hybrid nanoparticle system. J. Pharmacol. Exp. Ther. 317:1372 (2006).PubMedCrossRefGoogle Scholar
- 53.Taxol clinical pharmacology, RxList. http://www.rxlist.com/cgi/generic/paclitaxel_cp.htm.
- 57.W. P. McGuire. Paclitaxel in cancer treatment. Informa Health Care, Oxon, UK, 1995, p. 7.Google Scholar
- 58.M. G. Catalano, L. Costantino, N. Fortunati, O. Bosco, M. Pugliese, G. Boccuzzi, L. Berta, and R. Frairia. High energy shock waves activate 5’-aminolevulinic acid and increase permeability to Paclitaxel: antitumor effects of a new combined treatment on anaplastic thyroid cancer cells. Thyroid. 17:91 (2007).PubMedCrossRefGoogle Scholar