Transport of poly(amidoamine) dendrimers across Caco-2 cell monolayers: Influence of size, charge and fluorescent labeling.
OBJECTIVE
To investigate the transport of poly(amidoamine) (PAMAM) dendrimers with positive, neutral and negatively charged surface groups across Caco-2 cell monolayers.


METHODS
Cationic PAMAM-NH2 (G2 and G4), neutral PAMAM-OH (G2), and anionic PAMAM-COOH (G1.5-G3.5) dendrimers were conjugated to fluorescein isothiocyanate (FITC). The permeability of fluorescently labeled PAMAM dendrimers was measured in the apical-to-basolateral direction. 14C-Mannitol permeability was measured in the presence of unlabeled and FITC labeled PAMAM dendrimers. Caco-2 cells were incubated with the dendrimers followed by mouse anti-occludin or rhodamine phalloidin, and visualized using confocal laser scanning microscopy to examine tight junction integrity.


RESULTS
The overall rank order of PAMAM permeability was G3.5COOH > G2NH2 > G2.5COOH > G1.5COOH > G2OH. 14C-Mannitol permeability significantly increased in the presence of cationic and anionic PAMAM dendrimers with significantly greater permeability in the presence of labeled dendrimers compared to unlabeled. PAMAM dendrimers had a significant influence on tight junction proteins occludin and actin, which was microscopically evidenced by disruption in the occludin and rhodamine phalloidin staining patterns.


CONCLUSIONS
These studies demonstrate that enhanced PAMAM permeability is in part due to opening of tight junctions, and that by appropriate engineering of PAMAM surface chemistry it is possible to increase polymer transepithelial transport for oral drug delivery applications.
Kitchens
Kelly M
KM
Center for Nanomedicine and Cellular Delivery, Department of Pharmaceutical Sciences, University of Maryland, Baltimore, Baltimore, Maryland, USA.
Kolhatkar
Rohit B
RB
Swaan
Peter W
PW
Eddington
Natalie D
ND
Ghandehari
Hamidreza
H
D000199:Actins / Q000737:chemistry / Q000378:metabolism; D018938:Caco-2 Cells; D002463:Cell Membrane Permeability; D002470:Cell Survival / Q000187:drug effects; D050091:Dendrimers; D004563:Electrochemistry; D016650:Fluorescein-5-isothiocyanate; D005453:Fluorescence; D005456:Fluorescent Dyes; D006801:Humans; D008353:Mannitol / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D018613:Microscopy, Confocal; D062793:Occludin; D010316:Particle Size; D011073:Polyamines / Q000737:chemistry* / Q000493:pharmacokinetics* / Q000494:pharmacology; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism
D000199:Actins; D050091:Dendrimers; D005456:Fluorescent Dyes; D008565:Membrane Proteins; C568985:OCLN protein, human; D062793:Occludin; C568986:Ocln protein, mouse; C104700:PAMAM Starburst; D011073:Polyamines; D008353:Mannitol; D016650:Fluorescein-5-isothiocyanate