An ex vivo model of the rat trachea to study the effect of inhalable toxic compounds.
Different cell culture and organ systems are used to evaluate the physiological responses of the airways to the effects of carcinogenic [e.g., benzo(a)pyrene] and anticarcinogenic (e.g., retinoids) compounds on cellular growth and differentiation. However, in contrast to in vivo conditions dissociated epithelial cells or tracheal ring cultures are covered with medium. Therefore, we developed an ex vivo perfusion model enabling evaluation of morphology and metabolism of different compounds under near-physiological conditions. The trachea was surrounded with culture medium and perfused with air by means of a small animal respirator. To test the viability of the system under various experimental conditions tracheal probes were incubated with either retinoids (retinol 10(-5) mol/l; retinyl palmitate 10(-5) mol/l) or benzo(a)pyrene (10(-7) mol/l) for up to 7 days. At the end of the incubation period metabolites in the trachea and in the medium were measured by means of high-performance liquid chromatography. Samples were examined by light microscopy, and by scanning and transmission electron microscopy for cell morphology. Glycoconjugate expression was assessed by lectin histochemistry. Specimens incubated in a retinoid-supplemented medium revealed no alterations in the distribution of cell types and characteristics of the epithelial layer compared with tracheal biopsies assessed immediately after removal from the animals. Glycoconjugate patterns especially remained intact. Histological changes after incubation with benzo(a)pyrene resembled in vivo morphology of vitamin A-deficient rats. An important advantage of this in vitro model compared with common cell or organ cultures is the preservation of the original phenotype and environment of the tracheobronchial surface. In addition, carcinogenic substances, such as benzo(a)pyrene, can easily be applied by airway or through the medium.
Institut für Biologische Chemie und Ernährungswissenschaft der Universität Stuttgart-Hohenheim, Germany.
H K
HK
Biesalski
L
L
Engel
E
E
Stofft
A
A
Zschäbitz
D000818:Animals; D001580:Benzopyrenes / Q000494:pharmacology; D004224:Diterpenes; D037102:Lectins / Q000737:chemistry / Q000378:metabolism; D008854:Microscopy, Electron; D009844:Oligosaccharides / Q000737:chemistry; D009924:Organ Culture Techniques / Q000295:instrumentation / Q000379:methods; D011485:Protein Binding; D051381:Rats; D017208:Rats, Wistar; D000084562:Retinyl Esters; D014132:Trachea / Q000378:metabolism*; D014801:Vitamin A / Q000031:analogs & derivatives / Q000032:analysis / Q000378:metabolism; D014802:Vitamin A Deficiency; D015262:Xenobiotics / Q000633:toxicity*
D001580:Benzopyrenes; D004224:Diterpenes; D037102:Lectins; D009844:Oligosaccharides; D000084562:Retinyl Esters; D015262:Xenobiotics; D014801:Vitamin A; C014794:retinol palmitate