Engineering of Dendrimer Surfaces to Enhance Transepithelial Transport and Reduce Cytotoxicity
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Purpose. To evaluate the cytotoxicity, permeation, and transport mechanisms of PAMAM dendrimers and surface-modified cationic PAMAM dendrimers using monolayers of the human colon adenocarcinoma cell line, Caco-2.
Methods. Cytotoxicity was determined using the MTT assay. The effect of dendrimers on monolayer integrity was determined from measurements of transepithelial electrical resistance (TEER) and [14C]mannitol apparent permeability coefficient (Papp). The Papp of dendrimers through monolayers was measured in both the apical (A)-to-basolateral (B) and B→A directions at 4°C and 37°C and also in the presence and absence of ethylenediamine tetraacetic acid (EDTA) and colchicine.
Results. The cytotoxicity and permeation of dendrimers increased with both concentration and generation. The cytotoxicity of cationic dendrimers (G2, G3, G4) was greater than that of anionic dendrimers (G2.5, G3.5) but was reduced by conjugation with lauroyl chloride; the least cytotoxic conjugates were those with six attached lauroyl chains. At 37°C the Papp of cationic dendrimers was higher than that of anionic dendrimers and, in general, increased with the number of attached lipid chains. Cationic dendrimers decreased TEER and significantly increased the Papp of mannitol. Modified dendrimers also reduced TEER and caused a more marked increase in the Papp of mannitol. The Papp values of dendrimers and modified dendrimers were higher in the presence of EDTA, lower in the presence of colchicine, and lower at 4°C than at 37°C.
Conclusions. The properties of dendrimers may be significantly modified by surface engineering. Conjugation of cationic PAMAM dendrimers with lauroyl chloride decreased their cytotoxicity and increased their permeation through Caco-2 cell monolayers. Both PAMAM dendrimers and lauroyl-PAMAM dendrimer conjugates can cross epithelial monolayers by paracellular and transcellular pathways.
- D. A. Tomalia. Dendrimer molecules. Sci. Am. 272:62-66 (1995).
- A. D'Emanuele, D. Attwood, and R. Abu-Rmaileh. Dendrimers. In J. Swarbrick and J. C. Boylan (eds.), Encyclopedia of Pharmaceutical Technology, 2nd ed., update 3. Marcel Dekker, New York, 2002, pp. 1-21.
- J. C. Roberts, M. K. Bhalgat, and R. T. Zera. Preliminary biological evaluation of polyamidoamine (PAMAM) Starburst dendrimers. J. Biomed. Mater. Res. 30:53-65 (1996).
- N. Malik, R. Wiwattanapatapee, R. Klopsch, K. Lorenz, H. Frey, J. W. Weener, E. W. Meijer, W. Paulus, and R. Duncan. Dendrimers: Relationship between structure and biocompatibility in vitro, and preliminary studies on the biodistribution of 125I-labelled polyamidoamine dendrimers in vivo. J. Contr. Rel. 65:133-148 (2000).
- R. Jevprasesphant, J. Penny, R. Jalal, D. Attwood, N. B. McKeown, and A. D'Emanuele. The influence of surface modification on the cytotoxicity of PAMAM dendrimers. Int. J. Pharm. 252:263-266 (2003).
- A. T. Florence, T. Sakthivel, and I. Toth. Oral uptake and translocation of a polylysine dendrimer with a lipid surface. J. Contr. Rel. 65:253-259 (2000).
- R. Wiwattanapatapee, B. Carreno-Gomez, N. Malik, and R. Duncan. Anionic PAMAM dendrimers rapidly cross adult rat intestine in vitro: a potential oral delivery system. Pharm. Res. 17:991-998 (2000).
- M. El-Sayed, M. Ginski, C. Rhodes, and H. Ghandehari. Transepithelial transport of poly(amidoamine) dendrimers across Caco-2 cell monolayers. J. Contr. Rel. 81:355-365 (2002).
- P. Artursson. Epithelial transport of drugs in cell culture. I: A model for studying the passive diffusion of drugs over intestinal absorptive (Caco-2) cells. J. Pharm. Sci. 79:476-482 (1990).
- P. Artursson and R. T. Borchardt. Intestinal drug absorption and metabolism in cell cultures, Caco-2 and beyond. Pharm. Res. 14:1655-1658 (1997).
- R. T. Borchardt, H. E. Lane, B. H. Hirst, P. L. Smith, K. L. Audus, and A. Tsuji. Application of cell culture systems to the study of drug transport and metabolism. In A. C. Cuello and B. Collier (eds.), Pharmacological Sciences: Perspectives for Research and Therapy in the Late 1990s, Verlag, Basel, 1995, pp. 493-501.
- C. A. Bailey, P. Bryla, and A. W. Malick. The use of the intestinal epithelial cell culture model, Caco-2, in pharmaceutical development. Adv. Drug Deliv. Rev. 22:85-103 (1996).
- L.-S. L. Gan and D. R. Thakker. Applications of the Caco-2 model in the design and development of orally active drugs: elucidation of biochemical and physical barriers posed by the intestinal epithelium. Adv. Drug Deliv. Rev. 23:77-98 (1997).
- P. Artursson, K. Palm, and K. Luthman. Caco-2 monolayers in experimental and theoretical predictions of drug transport. Adv. Drug Deliv. Rev. 46:27-43 (2001).
- T. Lindmark, T. Nikkila, and P. Artursson. Mechanisms of absorption enhancement by medium chain fatty acids in intestinal epithelial Caco-2 cell monolayers. J. Pharmacol. Exp. Ther. 275:958-964 (1995).
- K. Higaki, T. Yata, M. Sone, K.-I. Ogawara, and T. Kimura. Estimation of absorption enhancement by medium-chain fatty acids in rat large intestine. Res. Commun. Mol. Pathol. Pharmacol. 109:231-240 (2001).
- Y. Kimura, Y. Hosoda, M. Shima, S. Adachi, and R. Matsuno. Physicochemical properties of fatty acids for assessing the threshold concentration to enhance the absorption of a hydrophilic substance. Biosci. Biotechnol. Biochem. 62:443-447 (1998).
- T. Lindmark, Y. Kimura, and P. Artursson. Absorption enhancement through intracellular regulation of tight junction permeability by medium chain fatty acids in Caco-2 cells. J. Pharmacol. Exp. Ther. 284:362-369 (1998).
- M. Tomita, M. Hayashi, and S. Awazu. Absorption-enhancing mechanism of EDTA, caprate, and decanoylcarnitine in Caco-2 cells. J. Pharm. Sci. 85:608-611 (1996).
- H. S. Thatte, K. R. Bridges, and D. E. Golan. Microtubule inhibitors differentially affect translational movement, cell surface expression, and endocytosis of transferrin receptors in K562 cells. J. Cell. Physiol. 160:345-357 (1994).
- S. L. Snyder and P. Z. Sobocinski. An improved 2,4,6-trinitrobenzenesulfonic acid method for the determination of amines. Anal. Biochem. 64:284-288 (1975).
- T. Mosmann. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J. Immun. Methods 65:55-63 (1983).
- G. A. Brazeau, S. Attia, S. Poxon, and J. A. Hughes. In vitro myotoxicity of selected cationic macromolecules used in non-viral gene delivery. Pharm. Res. 15:680-684 (1998).
- H. Yoo and R. L. Juliano. Enhanced delivery of antisense oligonucleotides with fluorophore-conjugated PAMAM dendrimers. Nucleic Acids Res. 28:4225-4231 (2000).
- V. H. L. Lee and J. J. Yang. Oral Drug Delivery. In A. M. Hillery, A. W. Lloyd, and J. Swarbrick (eds.), Drug Delivery and Targeting, Taylor and Francis, London, 2001, pp. 145-183.
- P. Artursson, T. Lindmark, S. S. Davis, and L. Illum. Effect of chitosan on the permeability of monolayers of intestinal epithelial cells (Caco-2). Pharm. Res. 11:1358-1361 (1994).
- G. Ranaldi, I. Marigliano, I. Vespignani, G. Perozzi, and Y. Sambuy. The effect of chitosan and other polycations on tight junction permeability in the human intestinal Caco-2 cell line. J. Nutr. Biochem. 13:157-167 (2002).
- P. Anderle, E. Niederer, W. Rubas, C. Hilgendorf, H. Spahn-Langguth, H. Wunderli-Allenspach, H. P. Merkle, and P. Langguth. P-glycoprotein (P-gp) mediated efflux in Caco-2 cell monolayers: the influence of culturing conditions and drug exposure on P-gp. J. Pharm. Sci. 87:757-762 (1998).
- C. H. M. Versantvoort, R. C. A. Ondrewater, E. Duizer, J. J. M. Van de Sandt, A. J. Gilde, and J. P. Groten. Monolayers of IEC-18 cells as an in vitro model for screening the passive transcellular and paracellular transport across the intestinal barrier: comparison of active and passive transport with the human colon carcinoma Caco-2 cell line. Environ. Toxicol. Pharmacol. 11:335-344 (2002).
- A. B. J. Noach, Y. Kurosaki, M. C. M. Blom-Roosemalen, A. G. de Boer, and D. D. Breimer. Cell-polarity dependent effect of chelation on the paracellular permeability of confluent Caco-2 cell monolayers. Int. J. Pharm. 90:229-237 (1993).
- C. B. Collares-Buzato, G. T. A. McEwan, M. A. Jepson, N. L. Simmons, and B. H. Hirst. Paracellular barrier and junctional protein distribution depend on basolateral extracellular Ca2+ in cultured epithelia. Biochim. Biophys. Acta 1222:147-158 (1994).
- Engineering of Dendrimer Surfaces to Enhance Transepithelial Transport and Reduce Cytotoxicity
Volume 20, Issue 10 , pp 1543-1550
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