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Leakiness and Size Exclusion of Paracellular Channels in Cultured Epithelial Cell Monolayers–Interlaboratory Comparison

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Abstract

Purpose

To determine and compare the paracellular characteristics of permeability (Papp) of Caco-2, MDCK, and 2/4/A1 cell lines.

Methods

The Papp data from 14 studies were analyzed by weighted nonlinear regression in terms of the paracellular parameters: porosity-pathlength (ε/δ), pore radius (R), and electrostatic potential drop (Δφ). Aqueous diffusivities, Daq, for the analysis, were empirically determined. The required hydrodynamic radii, rHYD, were estimated without knowledge of compound density. Mannitol iso-paracellular profiles allowed comparisons of “leakiness” across labs.

Results

Daq (37°C) was predicted as 9.9 × 10−5 MW−0.453; rHYD = (0.92 + 21.8 MW−1)·rSE, where rSE is the Stokes-Einstein radius. Values of pore radius ranged from 4.0(±0.1) to 18(±3) Å, with the 2/4/A1 indicating the largest pores. The ε/δ capacity factor ranged from 0.2 (±0.1) to 69 (±5) cm−1, with most values <1.5 cm−1. The average potential drop for Caco-2 models was Δφ Caco-2wt avg  = −43 ± 20 mV. The paracellular model predicted measured log Papp values with pooled r 2 = 0.93 and s = 0.17 (n = 108).

Conclusion

R and ε/δ are negatively correlated to a large extent. Papp can be rate-limited by either factor, with a wide range of possible combinations still indicating nearly constant leakiness for a given marker.

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ACKNOWLEDGEMENT

Helpful discussions with Dr. Kin Y. Tam (AstraZeneca, UK) and Dr. Kiyohiko Sugano (Pfizer, UK) are gratefully acknowledged. The long-term guiding influence of Prof. Norman F. H. Ho is deeply appreciated.

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Authors and Affiliations

  1. pION INC, 5 Constitution Way, Woburn, Massachusetts, 01801, USA

    Alex Avdeef

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  1. Alex Avdeef
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Correspondence to Alex Avdeef.

Additional information

Contribution number 27 in the PAMPA—a Drug Absorption in vitro Model series from pION. Ref. 26 is part 26 in the series.

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Avdeef, A. Leakiness and Size Exclusion of Paracellular Channels in Cultured Epithelial Cell Monolayers–Interlaboratory Comparison. Pharm Res 27, 480–489 (2010). https://doi.org/10.1007/s11095-009-0036-7

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