Correlation Between In Vivo and In Vitro Pulmonary Responses to Jet Propulsion Fuel-8 Using Precision-Cut Lung Slices and a Dynamic Organ Culture System

Abstract

In tissue slice models, interactions between the heterogeneous cell types comprising the lung parenchyma are maintained thus providing a controlled system for the study of pulmonary toxicology in vitro. However, validation of the model in vitro system must be affirmed. Previous reports, in in vivo systems, have demonstrated that Clara cells and alveolar type II cells are the targets following inhalation of JP-8 jet fuel. We have utilized the lung slice model to determine if cellular targets are similar following in vitro exposure to JP-8. Agar-filled adult rat lung explants were cored and precision cut, using the Brendel/Vitron tissue slicer. Slices were cultured on titanium screens located as half-cylinders in cylindrical Teflon cradles that were loaded into standard scintillation vials and incubated at 37 • C. Slices were exposed to JP-8 jet fuel (0.5 mg/ml, 1.0 mg/ml, and 1.5 mg/ml in medium) for up to 24 hours. We determined ATP content using a luciferin-luciferase bioluminescent assay. No significant difference was found between the JP-8 jet fuel doses or time points, when compared to controls. Results were correlated with structural alterations following aerosol inhalation of JP-8. As a general observation, ultrastructural evaluation of alveolar type II cells revealed an apparent increase in the number and size of surfactant secreting lamellar bodies that was JP-8 jet fuel-dose dependent. These results are similar to those observed following aerosol inhalation exposure. Thus, the lung tissue slice model appears to mimic in vivo effects of JP-8 and therefore is a useful model system for studying the mechanisms of lung injury following JP-8 exposure.

In vitrolung slices: a powerful approach for assessment of lung pathophysiology

“…For example, one group evaluated whether cellular targets of toxicity were similar after in vitro or in vivo exposure of lung to JP-8 jet fuel [27]. Ultrastructural evaluation of alveolar type II cells exposed as slices to the jet fuel showed a dose-dependent increase in the number and size of surfactantsecreting lamellar bodies; this finding was similar to the effect of in vivo exposure by aerosol inhalation [27].…”

“…For example, one group evaluated whether cellular targets of toxicity were similar after in vitro or in vivo exposure of lung to JP-8 jet fuel [27]. Ultrastructural evaluation of alveolar type II cells exposed as slices to the jet fuel showed a dose-dependent increase in the number and size of surfactantsecreting lamellar bodies; this finding was similar to the effect of in vivo exposure by aerosol inhalation [27]. Thus, in vitro exposure of lung tissue slices appears to reproduce the in vivo effects of JP-8 exposure, making slices a useful model system for studying the mechanisms of lung injury after exposure to this substance [27].…”

“…The gentle rolling motion of the vials allows the tissue slice to be exposed on both sides to air or the liquid medium, thus providing exposure to xenobiotics through the liquid or gaseous phase. This approach has been used to study cyclosporine and its derivatives [23], 7-ethoxycoumarin and paraquat [24,25], complex aerosols [26], benzo[a] pyrene [13] and jet fuel [27]. Tissues from several different species, including humans, have been studied in this manner [6].…”

Translational Research in Pharmacology and Toxicology Using Precision-Cut Tissue Slices

“…To model inhalation toxicology, the dynamic organ culture methods as described above have been successfully applied to expose lung slices to aerosols and tobacco smoke. In general, precision-cut lung slices of animals can respond to toxic compounds in a comparable way as in vivo, showing dose-dependent toxicity of chemicals like 3-methylindole, 1-nitronaphtalene and paraquat, 76 genotoxicity of formalin and ethyl methane sulfonate, 77 covalent binding to proteins of agaritine 78 apoptosis, proinammatory response and cytokine production by diesel exhaust, 79 endothelial dysfunction and IL1b release by cigarette smoke and nicotine, 80-82 surfactant lamellar body secretion by jet propulsion uid, 83 inammatory reactions and loss of viability by ozone and nitrogen dioxide, 84 and ATP and glutathione decrease by acrolein. 85 However, the number of toxicity studies in human lung slices is relatively limited.…”

Lung Organoid Culture to Study Responses to Viruses

“…As a result of this development, combined with the ongoing development of live tissue microtomes, there has been a significant increase in the use of precision cut organ culture models in several applications. These include studies of lung exposure to asbestos, cadmium chloride, benzopyrene, bleomycin, 3-methylindole, 1-napthalene, paraquat, diesel exhaust, jet fuel, and cigarette smoke [6][7][8][9][10][11][12][13] . Also, studies of lung metabolism of pharmaceuticals have been examined, including the effect of drugs and irritants on airway bronchoconstriction, which will be reviewed in another chapter of this book.…”