Endoscopic biopsies in Ussing chambers evaluated for studies of macromolecular permeability in the human colon.
OBJECTIVE
Studies of mucosal permeability to protein antigens in humans are limited to in vitro techniques. The use of surgical specimens for such studies has major shortcomings. Endoscopic biopsies in Ussing chambers have been introduced as a means of studying secretion and transepithelial permeability, but have not been evaluated for studies of protein antigen uptake in human intestine.


METHODS
Standard forceps biopsies from the sigmoid colon of 24 healthy volunteers were mounted in Ussing chambers with an exposed tissue area of 1.76 mm2. 51Cr-EDTA (paracellular probe) and horseradish peroxidase (HRP; 45 kDa protein antigen) were used as permeability markers. Mucosal permeability, electrophysiology, histology and energy contents of the biopsies were studied over time. To evaluate the ability of the technique to detect permeability changes, the mucosa was modulated with capric acid, a medium-chain fatty acid, known to affect tight junctions.


RESULTS
In the Ussing chamber the mucosal biopsies were viable for 160 min with stable levels of ATP and lactate, and only minor changes in morphology. Steady-state permeability with low variability was seen for both markers during the 30-90 min period. Exposure to capric acid induced a rapid decrease in short-circuit current (Isc) and a slower reversible decrease in transepithelial resistance (TER), as well as an increased permeability to 51Cr-EDTA and HRP.


CONCLUSIONS
Endoscopic biopsies of human colon are viable in Ussing chambers and are reliable tools for studies of mucosal permeability to protein antigens. The technique offers a broad potential for studies of mucosal function in the pathophysiology of human gastrointestinal diseases.
Wallon
Conny
C
Clinical Research Centre, University of Linköping, Sweden. conny.wallon@lio.se
Braaf
Ylva
Y
Wolving
Mats
M
Olaison
Gunnar
G
Söderholm
Johan D
JD
D000255:Adenosine Triphosphate / Q000378:metabolism; D000328:Adult; D001706:Biopsy; D002860:Chromium Radioisotopes; D012809:Colon, Sigmoid / Q000166:cytology* / Q000378:metabolism; D003652:Decanoic Acids / Q000494:pharmacology; D004492:Edetic Acid; D005260:Female; D006801:Humans; D007413:Intestinal Mucosa / Q000187:drug effects / Q000378:metabolism; D019344:Lactic Acid / Q000378:metabolism; D046911:Macromolecular Substances / Q000493:pharmacokinetics; D008297:Male; D010539:Permeability; D012812:Sigmoidoscopy
D002860:Chromium Radioisotopes; D003652:Decanoic Acids; D046911:Macromolecular Substances; D019344:Lactic Acid; C031071:decanoic acid; D000255:Adenosine Triphosphate; D004492:Edetic Acid