In vitro drug absorption models. I. Brush border membrane vesicles, isolated mucosal cells and everted intestinal rings: characterization and salicylate accumulation.

Brush border membrane vesicles, isolated mucosal cells and everted rings from rat intestine were compared for their suitability for drug uptake studies. Vesicles from brush border membranes were judged to be metabolically and morphologically functional on the basis of biochemical and microscopic criteria. With the use of a collagenase-vascular-perfusion method, populations of villus, mid villus and crypt cells were separated. An alternative approach that is based on an EDTA-dissociation procedure afforded fractions enriched in villus and crypt cells. Although several enzymatic and metabolic activities of these two cell preparations were comparable, cell viability based on the Trypan Blue dye exclusion test, ultrastructural appearance and glucose uptake more closely conformed to in vivo values for cells isolated according to the EDTA-dissociation method. These cells were chosen as a model for drug transport investigation. The morphological and functional integrity of everted rings was verified by histological examination, extracellular space estimation and assessment of glucose transport ability. Sodium salicylate uptake studies using brush border membrane vesicles and isolated mucosal cells were highly variable, whereas everted segments exhibited good reproducibility in uptake experiments. Time dependence of salicylate uptake was demonstrated with membrane vesicles and everted rings. Time dependence was not observed in mucosal cell uptake studies, probably because of the time required to separate the cells from the incubation solution. Based on ease of preparation, technical aspects of in vitro incubation and reproducibility of results, everted intestinal rings were considered to be a good potential model for in vivo drug absorption. Brush border membrane vesicles were generally regarded as unacceptable because of variations after storage and between experiments. Isolated cells offered certain advantages, but the utility of cells as an in vitro model remains equivocal.