Effectiveness and toxicity screening of various absorption enhancers using Caco-2 cell monolayers.
We studied the enhancing and toxic effects of five different absorption enhancers on the transport of FITC-dextran with an average molecular weight of 4000 (FD-4) across Caco-2 cell monolayers, and their enhancing effects were also compared with those in rat intestine. The enhancing and cytotoxic properties of these enhancers were characterized using the following tests: measurement of the permeability coefficients of FD-4 and the transepithelial electrical resistance (TEER) in Caco-2, the release of cytosolic lactate dehydrogenase (LDH) and intracellular mitochondrial dehydrogenase (MDH) activity. All the absorption enhancers increased the permeability of FD-4 across Caco-2 cell monolayers and a good relationship was observed between the enhancement and their toxic effects. However, EDTA and Na-Cap were effective for improving the transport of FD-4 across Caco-2 cells without serious cytotoxicity. At concentrations with low cytotoxicity, various absorption enhancers exihibited reversible effects on the TEER values in Caco-2 cell monolayers, except for 50 mM sodium salicylate (Na-Sal). Moreover, we obtained a good correlation between the enhancement of these enhancers in Caco-2 cell monolayers and in rat large intestine. This finding indicated that the effectiveness of absorption enhancers in the Caco-2 monolayer system was similar to an in vivo rat system. Therefore, the screening system using Caco-2 cell monolayers is useful for examining the effectiveness and toxicity of absorption enhancers.
Quan
Y S
YS
Department of Biopharmaceutics, Kyoto Pharmaceutical University, Japan.
Hattori
K
K
Lundborg
E
E
Fujita
T
T
Murakami
M
M
Muranishi
S
S
Yamamoto
A
A
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D018938:Caco-2 Cells; D003652:Decanoic Acids / Q000494:pharmacology / Q000633:toxicity; D003911:Dextrans / Q000378:metabolism*; D004492:Edetic Acid / Q000494:pharmacology / Q000633:toxicity; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives* / Q000378:metabolism; D006000:Glycocholic Acid / Q000494:pharmacology / Q000633:toxicity; D006801:Humans; D007408:Intestinal Absorption / Q000187:drug effects*; D007420:Intestine, Large / Q000187:drug effects / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008320:Maltose / Q000031:analogs & derivatives / Q000494:pharmacology / Q000633:toxicity; D008928:Mitochondria / Q000201:enzymology; D051381:Rats; D017208:Rats, Wistar; D012980:Sodium Salicylate / Q000494:pharmacology / Q000633:toxicity
D003652:Decanoic Acids; D003911:Dextrans; C015219:fluorescein isothiocyanate dextran; C109906:n-lauryl beta-d-maltopyranoside; C031071:decanoic acid; D008320:Maltose; D004492:Edetic Acid; D007770:L-Lactate Dehydrogenase; D006000:Glycocholic Acid; D016650:Fluorescein-5-isothiocyanate; D012980:Sodium Salicylate