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Transport of Lipophilic Drug Molecules in a New Mucus-Secreting Cell Culture Model Based on HT29-MTX Cells

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Abstract

Purpose. A new mucus-secreting in vitro drug absorption model based on monolayers of goblet-cell like sub-clones of the human colon carcinoma cell line HT29 obtained by methotrexate (MTX) treatment was investigated.

Methods. Twelve sub-clones were isolated and characterized by light microscopy (LM), transelectron microscopy (TEM), confocal laser scanning microscopy (CLSM), transepithelial electrical resistance (TEER) and the transport of a paracellular marker FITC-Dextran (Mw 4400) (FD-4).

Results. Significant differences of microscopical appearance, TEER-values and permeability of FD-4 between the sub-clones were evident. However, two of them, namely MTX-D1 and MTX-E12, formed tight confluent monolayers with a thick mucus-layer on the apical surface. They were used to compare the apparent permeability coefficient (Papp) of a series of lipophilic drugs, which should be affected by the mucus-layer, namely barbiturates (barbituric acid, barbital, phenobarbital, methylphenobarbital and heptabarbital) and testosterone, as a reference, to mucus-free Caco-2 cells. The permeability of drugs with a partition coefficient (log P) > 1 was decreased in the mucus-producing cell lines. Testosterone, the most lipophilic compound, showed a decrease of up to 43%.

Conclusions. We demonstrated that the mucus layer is a significant barrier to drug absorption for lipophilic drugs. In conclusion, our model may serve as a suitable in-vitro cell culture model to study the influence of the mucus layer on drug diffusion.

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Authors and Affiliations

  1. Department of Pharmaceutics and Biopharmacy, University of Marburg, Ketzerbach 63, 35032, Marburg, Germany

    Isabel Behrens & Thomas Kissel

  2. Division of Pharmaceutics and Biopharmacy, Uppsala University, Box 500, SE-75123, Uppsala, Sweden

    Patric Stenberg & Per Artursson

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  1. Isabel Behrens
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  2. Patric Stenberg
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  4. Thomas Kissel
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Behrens, I., Stenberg, P., Artursson, P. et al. Transport of Lipophilic Drug Molecules in a New Mucus-Secreting Cell Culture Model Based on HT29-MTX Cells. Pharm Res 18, 1138–1145 (2001). https://doi.org/10.1023/A:1010974909998

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