Feasibility of the Ussing chamber technique for the determination of in vitro jejunal permeability of passively absorbed compounds in different animal species

Abstract

The aim of this study was to assess the feasibility of the Ussing chamber technique for the determination of the jejunal permeability of passively absorbed, high permeability model compounds (acetaminophen and ketoprofen) in different animal species. Additionally, electrophysiological measurements and histological examination of pre- and post-incubation tissue specimens were performed. Apparent permeability coefficients of turkey and dog jejunum were low and highly variable due to tissue fragility caused by differences in thickness of the remaining intestinal layers after stripping and resulting in severe damage. Pig and horse jejunum were markedly more suitable for permeability determinations and mild signs of deterioration were noticed after 120 min of incubation. Transepithelial electrical resistance and potential difference did not correlate well with the observed tissue damage. From these data, the Ussing chamber technique appears to allow for permeability measurements within a species, but seems unsuitable for interspecies permeability comparison. However, further validation of the method with low permeability compounds and actively transported compounds is needed.

Evaluation of endoscopically obtained duodenal biopsy samples from cats and dogs in an adapter-modified Ussing chamber

“…However, the resistance of stripped canine jejunum has previously been reported by Neirinckx et al . [24] (250.9 ± 109.2 Ω·cm 2 ), and, while the standard deviation in these samples was about 50%, similar variations of tissue resistance were reported for the small intestine of rats (20 ~50 Ω·cm 2 ) [27,29]. Since we transferred only the thickest and most intact looking biopsy samples into the Ussing chamber, it nearly always evaluated the full thickness biopsies.…”

Applying Biopharmaceutical Classification System (BCS) Criteria to Predict Oral Absorption of Drugs in Dogs: Challenges and Pitfalls

“…Because there are no commercially available canine cell lines (21), in the absence of in vitro BNot similar^was defined as outside this range permeability data, we have relied upon LogP values as the basis for estimating a canine BCS permeability cutoff value. Our selection of LogP values was founded upon published reports where LogP was used as a surrogate for human permeability studies (3)(4)(5)(6).…”

“…The Ussing chamber technique, which has been evaluated for other veterinary species (20), has not been applicable for canine studies (21) because of the fragility of the tissue. Membrane damage that occurs prevents permeability measurements using this technique in dogs (21). Therefore, without the availability of these in vitro tools, other data must be used to predict permeability and apply BCS criteria for oral drugs administered to dogs.…”

Establishment of Novel Prediction System of Intestinal Absorption in Humans Using Human Intestinal Tissues

“…The advantage offered by Ussing chamber studies over cultured cell lines and PAMPA is the maintenance of the complex cellular heterogeneity and morphology of the mammalian intestine and structure of epithelial cells, which are not entirely mimicked by the relatively homogeneous cellular monolayer of Caco‐2 cells and the artificial membrane of PAMPA. The presence of an extended myriad of transport proteins and drug metabolism enzymes, as well as an intact acid microclimate mucous layer adjacent to the villus tip of enterocytes, results in an experimental system that gives a more complete picture regarding the process that may impact drug absorption5–7; however, species differences in permeability and metabolic capability are already known,8–11 suggesting that predictions based on studies using other species' tissues would not be necessarily in accordance with the absorbability of drugs in humans 12–15. The usage of human tissue, therefore, has a great advantage for more precise prediction of drug absorbability in humans.…”