Transepithelial transport of poly(amidoamine) dendrimers across Caco-2 cell monolayers.

The objective of this study was to investigate the influence of physiochemical parameters (such as size, molecular weight, molecular geometry, and number of surface amine groups) of poly (amidoamine) (PAMAM) dendrimers, on their permeability across Caco-2 cell monolayers. The permeability of a series of PAMAM dendrimers, generations 0-4 (G0-G4), was investigated across Caco-2 cell monolayers in both the apical to basolateral (AB) and basolateral to apical (BA) directions. The influence of PAMAM dendrimers on the integrity, paracellular permeability, and viability of Caco-2 cell monolayers was also monitored by measuring the transepithelial electrical resistance (TEER), mannitol permeability, and leakage of lactate dehydrogenase (LDH) enzyme, respectively. G0, G1 and G2 demonstrated similar AB permeabilities, which were moderate several fold higher than the AB permeability of higher generations. The AB and BA permeability of G0-G4 typically increased with the increase in donor concentration and incubation time. Permeability values are not reported at generations, concentrations or incubation times that the dendrimers were toxic to Caco-2 cells. TEER values decreased and mannitol permeability increased as a function of donor concentration, incubation time, and generation number. LDH results for G3 and G4 indicate that Caco-2 cell viability was reduced with increasing donor concentration, incubation time, and generation number. The appreciable permeability of G0-G2, coupled with their nontoxic effects on Caco-2 cells, suggest their potential as water-soluble polymeric drug carriers for controlled oral drug delivery.

[1]  William A. Goddard,et al.  Starburst Dendrimers: Molecular‐Level Control of Size, Shape, Surface Chemistry, Topology, and Flexibility from Atoms to Macroscopic Matter , 1990 .

[2]  Malcolm Rowland,et al.  Comparison of HT29-18-C1 and Caco-2 Cell Lines as Models for Studying Intestinal Paracellular Drug Absorption , 1996, Pharmaceutical Research.

[3]  R T Borchardt,et al.  Paracellular diffusion in Caco-2 cell monolayers: effect of perturbation on the transport of hydrophilic compounds that vary in charge and size. , 1997, Journal of pharmaceutical sciences.

[4]  J. Hughes,et al.  In Vitro Myotoxicity of Selected Cationic Macromolecules Used in Non-Viral Gene Delivery , 1998, Pharmaceutical Research.

[5]  Ruth Duncan,et al.  Anionic PAMAM Dendrimers Rapidly Cross Adult Rat Intestine In Vitro: A Potential Oral Delivery System? , 2000, Pharmaceutical Research.

[6]  J. Kopeček,et al.  Size-dependent permeability of hydrophilic probes across rabbit colonic epithelium. , 1997, The Journal of pharmacology and experimental therapeutics.

[7]  D. Tomalia.  Starburst/Cascade Dendrimers: Fundamental building blocks for a new nanoscopic chemistry set , 1994 .

[8]  Per Artursson,et al.  Selective Paracellular Permeability in Two Models of Intestinal Absorption: Cultured Monolayers of Human Intestinal Epithelial Cells and Rat Intestinal Segments , 1993, Pharmaceutical Research.

[9]  R. Duncan,et al.  PAMAM dendrimers as a potential oral drug delivery system: Uptake by everted rat intestinal sacs in‐vitro , 1998 .

[10]  Gerrit Borchard,et al.  The potential of mucoadhesive polymers in enhancing intestinal peptide drug absorption. III: Effects of chitosan-glutamate and carbomer on epithelial tight junctions in vitro , 1996 .

[11]  G. Amidon,et al.  Peptide-based Drug Design, Controlling Transport and Metabolism , 1995 .

[12]  H. Junginger,et al.  N-Trimethyl Chitosan Chloride as a Potential Absorption Enhancer Across Mucosal Surfaces: In Vitro Evaluation in Intestinal Epithelial Cells (Caco-2) , 1997, Pharmaceutical Research.

[13]  A. Seelig,et al.  Substrate recognition by P-glycoprotein and the multidrug resistance-associated protein MRP1: a comparison. , 2000, International journal of clinical pharmacology and therapeutics.

[14]  D. Breimer,et al.  Cell-polarity dependent effect of chelation on the paracellular permeability of confluent caco-2 cell monolayers , 1993 .

[15]  R L Vessella,et al.  Biotin reagents for antibody pretargeting. 3. Synthesis, radioiodination, and evaluation of biotinylated starburst dendrimers. , 1998, Bioconjugate chemistry.

[16]  J. Polli,et al.  Transport of poly amidoamine dendrimers across Madin-Darby canine kidney cells. , 2001, International journal of pharmaceutics.

[17]  H. Junginger,et al.  Comparison of the effect of different chitosan salts and N-trimethyl chitosan chloride on the permeability of intestinal epithelial cells (Caco-2). , 1998, Journal of controlled release : official journal of the Controlled Release Society.

[18]  H. Junginger,et al.  Enhancement of paracellular drug transport with highly quaternized N-trimethyl chitosan chloride in neutral environments: in vitro evaluation in intestinal epithelial cells (Caco-2). , 1999, Journal of pharmaceutical sciences.

[19]  H. Junginger,et al.  N-Trimethylated Chitosan Chloride (TMC) Improves the Intestinal Permeation of the Peptide Drug Buserelin In Vitro (Caco-2 Cells) and In Vivo (Rats) , 2004, Pharmaceutical Research.

[20]  H. Ghandehari,et al.  Extravasation of Poly(amidoamine) (pamam) Dendrimers Across Microvascular Network Endothelium , 2004, Pharmaceutical Research.

[21]  J. Robinson,et al.  Transcellular and Lipophilic Complex-Enhanced Intestinal Absorption of Human Growth Hormone , 1999, Pharmaceutical Research.

[22]  E. W. Meijer,et al.  Dendrimers: relationship between structure and biocompatibility in vitro, and preliminary studies on the biodistribution of 125I-labelled polyamidoamine dendrimers in vivo. , 2000, Journal of controlled release : official journal of the Controlled Release Society.