Acute Cigarette Smoke Exposure Induces Cytotoxicity And Inflammation In Living Tissue Of Precision-Cut Lung Slices

Abstract

Chronic obstructive pulmonary disease (COPD) is a severe lung disease with high mortality and increasing prevalence. It is characterized by profound damage of lung parenchyma due to cigarette smoke-induced stress. Lipopolysaccharide (LPS) is a widely used model compound which does not directly induce oxidative stress due to lacking radicals. It therefore can not reflect cigarette smokeinduced toxicity in the lung. The aim of this study is to establish precision-cut lung slices (PCLS) as a relevant toxicity model by using cigarette smoke and cigarette smoke condensate in comparison to LPS as model compounds. Murine PCLS were prepared and exposed submerged to LPS and cigarette smoke condensate, and to cigarette mainstream smoke using the P.R.I.T. ® air-liquid interface (ALI) culturing and exposure system. Induced toxicity was assessed by LIVE/DEAD ® vitality staining and determination of metabolic activity using the WST-1 assay. Proinflammatory immune responses connected to toxic and subtoxic doses were quantified using ELISA. Additionally, therapeutic intervention with dexamethasone and roflumilast was assessed.

Murine precision-cut lung slices exhibit acute responses following exposure to gasoline direct injection engine emissions

“…Comparison of the airway responses across ex vivo and in vivo exposure models suggests similar trends: diesel engine exhaust was found to induce increased protein expression of the pro-inflammatory marker TNFα in both PCLS prepared from rat lung tissue exposed at an air-liquid interface and in lavage samples collected from rat lungs following inhalation exposure (Morin et al, 2008). Studies have reported increased protein expression of TNFα in response to particulate pollutants, such as engineered silica nanoparticles (Neuhaus et al, 2013), cigarette smoke (Lauenstein et al, 2012) and diesel engine exhaust particulate matter (Bion et al, 2002).…”