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| IJCEM Copyright © 2008-All rights reserved. Published by e-Century Publishing Corporation, Madison, WI 53711 |
Int J Clin Exp Med 2010;3(3):223-232
Original Article
A cellular model to mimic exhaled cigarette smoke-induced lung microvascular
endothelial cell injury and death
Jianliang Zhang, Noah Juedes, Vikram M. Narayan, Bingfang Yue, Alan L. Rockwood, Nadia L. Palma, Jawaharlal M. Patel
Department of Medicine, University of Florida College of Medicine, Gainesville, FL 32610, USA; Research Service, Malcom Randall
Department of Veterans Affairs Medical Center, Gainesville, FL 32608, USA; ARUP Institute for Clinical and Experimental Pathology, Salt
Lake City, UT 84108, USA; and Department of Pathology, University of Utah, Salt Lake City, UT 84132, USA
Received June 26, 2010; accepted July 29, 2010; available online July 31, 2010
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 cytotoxic 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. (IJCEM1006003).
Key words: Cellular model, secondhand smoke, chronic lung diseases, lung microvascular endothelial cells, passive
smoking-associated lung diseases
Full Text PDF
Address all correspondence to:
Jianliang Zhang, PhD
Pulmonary Division, MSB Rm M452
Department of Medicine
University of Florida College of Medicine
1600 S. W. Archer Road, Gainesville
FL 32610-0225, USA.
Tel: 353-376-1611, Ext. 6265, Fax: 352-374-6170
E-mail: Zhangjl@medicine.ufl.edu
Original Article
A cellular model to mimic exhaled cigarette smoke-induced lung microvascular
endothelial cell injury and death
Jianliang Zhang, Noah Juedes, Vikram M. Narayan, Bingfang Yue, Alan L. Rockwood, Nadia L. Palma, Jawaharlal M. Patel
Department of Medicine, University of Florida College of Medicine, Gainesville, FL 32610, USA; Research Service, Malcom Randall
Department of Veterans Affairs Medical Center, Gainesville, FL 32608, USA; ARUP Institute for Clinical and Experimental Pathology, Salt
Lake City, UT 84108, USA; and Department of Pathology, University of Utah, Salt Lake City, UT 84132, USA
Received June 26, 2010; accepted July 29, 2010; available online July 31, 2010
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 cytotoxic 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. (IJCEM1006003).
Key words: Cellular model, secondhand smoke, chronic lung diseases, lung microvascular endothelial cells, passive
smoking-associated lung diseases
Full Text PDF
Address all correspondence to:
Jianliang Zhang, PhD
Pulmonary Division, MSB Rm M452
Department of Medicine
University of Florida College of Medicine
1600 S. W. Archer Road, Gainesville
FL 32610-0225, USA.
Tel: 353-376-1611, Ext. 6265, Fax: 352-374-6170
E-mail: Zhangjl@medicine.ufl.edu
