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Transport of Poly(Amidoamine) Dendrimers across Caco-2 Cell Monolayers: Influence of Size, Charge and Fluorescent Labeling

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Abstract

Purpose

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.

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Abbreviations

FITC:

Fluorescein isothiocyanate

G:

Generation

HBSS:

Hank’s balanced salt solution

PAMAM:

Poly(amidoamine)

TEER:

Transepithelial electrical resistance

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Acknowledgments

The authors would like to acknowledge Amy Foraker for her assistance with the confocal microscopy studies. Kelly Kitchens received financial support for this research by a pre-doctoral National Research Service Award from the National Institute of General Medical Sciences (F31-GM67278).

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Authors and Affiliations

  1. Center for Nanomedicine and Cellular Delivery, Department of Pharmaceutical Sciences, University of Maryland, Baltimore, Baltimore, Maryland, USA

    Kelly M. Kitchens, Rohit B. Kolhatkar, Peter W. Swaan, Natalie D. Eddington & Hamidreza Ghandehari

  2. Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, 20 Penn Street, HSFII Room 625, Baltimore, Maryland, 21201-1075, USA

    Hamidreza Ghandehari

Authors
  1. Kelly M. Kitchens
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  2. Rohit B. Kolhatkar
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  3. Peter W. Swaan
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  4. Natalie D. Eddington
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  5. Hamidreza Ghandehari
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Correspondence to Hamidreza Ghandehari.

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Kitchens, K.M., Kolhatkar, R.B., Swaan, P.W. et al. Transport of Poly(Amidoamine) Dendrimers across Caco-2 Cell Monolayers: Influence of Size, Charge and Fluorescent Labeling. Pharm Res 23, 2818–2826 (2006). https://doi.org/10.1007/s11095-006-9122-2

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  • DOI: https://doi.org/10.1007/s11095-006-9122-2

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