title: A cellular model to mimic exhaled cigarette smokeinduced lung microvascular endothelial cell injury and death.
abstract: Tobacco smoke exhaled from smokers is a key component of secondhand smoke, contributing to lung alveolar wall destruction seen in chronic lung diseases. Although mainstream and sidestream tobacco smoke are cyto-toxic to lung cells, it is unclear whether exhaled smoke induces lung cell injury or even death. We sought to establish an in vitro model to examine the effects of exhaled smoke on lung cells. Phosphate-buffered saline-conditioned cigarette smoke (CCS) derived from a blow-by system was used to mimic exhaled tobacco smoke exposure. Exposure of medium to CCS leads to dose-dependent increases in nicotine/cotinine levels. Scanning spectrophotometric analysis of the CCS-exposed medium reveals an absorption peak at 290 nm wavelength. The OD values at 290 nm are correlated with nicotine levels in the exposed medium, indicating that a simple measurement of OD at 290 nm can be used to monitor CCS exposure. Tobacco smoke contacts the microvascular endothelium located at lung alveoli, before it enters the blood stream. Hence, human lung microvascular endothelial cells (hMVEC) were exposed to CCS and assessed for cell injury and death. Exposure of hMVEC to CCS equivalent to burning 12-16 cigarettes leads to increased LDH release from the cells into the medium. This suggests that CCS can induce lung cell injury. CCS at a low level increases cell growth, whereas the high level of CCS decreases cell viability. In addition, CCS exposure induces cell detachment and morphological changes. Our results demonstrate that exposure of buffer-conditioned mainstream cigarette smoke leads to increased nicotine/cotinine levels and cell injury/death, which may contribute to the pathophysiology of passive smoking-associated lung diseases.
authors.forename: Jianliang
authors.initials: J
authors.lastname: Zhang
authors.forename: Noah
authors.initials: N
authors.lastname: Juedes
authors.forename: Vikram M
authors.initials: VM
authors.lastname: Narayan
authors.forename: Bingfang
authors.initials: B
authors.lastname: Yue
authors.forename: Alan L
authors.initials: AL
authors.lastname: Rockwood
authors.forename: Nadia L
authors.initials: NL
authors.lastname: Palma
authors.forename: Jawaharlal M
authors.initials: JM
authors.lastname: Patel
references.citation: Indian J Chest Dis Allied Sci. 2006 Jan-Mar;48(1):23-9
references.pmid: 16482948
references.citation: Chest. 2000 May;117(5 Suppl 1):241S-2S
references.pmid: 10843927
references.citation: Am J Respir Cell Mol Biol. 1996 Sep;15(3):410-9
references.pmid: 8810647
references.citation: BMJ. 2003 Aug 30;327(7413):502; author reply 504-5
references.pmid: 12946981
references.citation: Biomed Environ Sci. 2004 Jun;17(2):121-34
references.pmid: 15386938
references.citation: Cancer Epidemiol Biomarkers Prev. 2009 Dec;18(12):3415-20
references.pmid: 19959690
references.citation: Respir Res. 2006 Mar 30;7:53
references.pmid: 16571143
references.citation: Mech Ageing Dev. 2002 Mar 31;123(6):613-25
references.pmid: 11850025
references.citation: BMC Pulm Med. 2006 Jun 06;6:12
references.pmid: 16756671
references.citation: Carcinogenesis. 2001 Mar;22(3):375-80
references.pmid: 11238175
references.citation: Am J Respir Cell Mol Biol. 2003 Nov;29(5):562-70
references.pmid: 12748058
references.citation: Am J Physiol Cell Physiol. 2002 Oct;283(4):C1080-9
references.pmid: 12225972
references.citation: J Occup Environ Med. 2001 Aug;43(8):687-93
references.pmid: 11515251
references.citation: Am J Respir Crit Care Med. 1995 Nov;152(5 Pt 1):1666-72
references.pmid: 7582312
references.citation: Indian J Otolaryngol Head Neck Surg. 1998 Jul;50(3):230-2
references.pmid: 23119424
references.citation: Br Med J. 1980 Apr 5;280(6219):972-6
references.pmid: 7417765
references.citation: J Appl Physiol (1985). 1997 Jan;82(1):3-12
references.pmid: 9029192
references.citation: Free Radic Biol Med. 2003 Feb 1;34(3):345-55
references.pmid: 12543250
references.citation: Mol Genet Metab. 2001 Jan;72(1):82-8
references.pmid: 11161833
references.citation: Am J Pathol. 2006 Oct;169(4):1155-66
references.pmid: 17003475
references.citation: AJR Am J Roentgenol. 1986 Feb;146(2):233-7
references.pmid: 3484568
references.citation: Am J Physiol Lung Cell Mol Physiol. 2001 Aug;281(2):L509-16
references.pmid: 11435227
references.citation: Environ Health. 2005 May 12;4(1):7
references.pmid: 15890079
references.citation: Am J Respir Cell Mol Biol. 2003 Jul;29(1):88-97
references.pmid: 12600822
references.citation: Methods Mol Biol. 2010;603:389-98
references.pmid: 20077091
references.citation: J Lab Clin Med. 1988 Jan;111(1):15-27
references.pmid: 3275730
references.citation: Am J Respir Crit Care Med. 2006 Aug 15;174(4):428-36
references.pmid: 16709937
references.citation: Handb Exp Pharmacol. 2009;(192):29-60
references.pmid: 19184645