Toxicological Sciences, Vol 51, 126-134, Copyright © 1999 by Society of Toxicology
JM Antonini, TG Charron, JR Roberts, J Lai, TL Blake and RA Rogers
Laser scanning confocal microscopy (LSCM) allows us to simultaneously
quantitate the degree of lung fibrosis and distinguish various pathological
lesions of intact lung tissue. Lucifer Yellow has been shown an ideal
fluorescent stain to examine the connective tissue matrix components of
embedded lung tissue with LSCM. We evaluated the use of LSCM in
quantitating lung fibrosis and compared this procedure with the more
traditional method of assessing fibrosis by measuring hydroxyproline, a
biochemical assay of collagen. CD/VAF rats were intratracheally dosed with
silica (highly fibrogenic), Fe2O3 (non- fibrogenic), and saline (vehicle
control) at a high dose of 10-mg/100 g body weight. At 60 days
post-instillation, the left lung was dissolved in 6 M HCl and assayed for
hydroxyproline. Silica induced increases of 58% and 94% in hydroxyproline
content over the Fe2O3 and control groups, respectively. The right lung
lobes were fixed, sectioned into blocks, dehydrated, stained with Lucifer
Yellow (0.1 mg/ml), and embedded in Spurr plastic. Using LSCM and
ImageSpace software, the tissue areas of ten random scans from ten blocks
of tissue for each of the three groups were measured, and three-dimensional
reconstructions of random areas of lung were generated. The silica group
showed increases of 57% and 60% in the lung areas stained by Lucifer Yellow
over the Fe2O3 and control groups, respectively. Regression analysis of
hydroxyproline vs. lung tissue area demonstrated a significant positive
correlation (p < 0.05) with a correlation coefficient of 0.91.
Histological analysis of right lung tissue revealed a marked degree of
granulomatous interstitial pneumonitis for the silica group, which was
absent in the Fe2O3 and control groups. No significant differences (p <
0.05) in hydroxyproline content and measured tissue area were observed
between the Fe2O3 and control groups. LSCM, and its associated advanced
image analysis and three-dimensional capabilities, is an alternative method
to both quickly quantitate and examine fibrotic lung disease without
physical disruption of the tissue specimen.
ARTICLES
Application of laser scanning confocal microscopy in the analysis of particle-induced pulmonary fibrosis
Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Morgantown, West Virginia 26505, USA. jga6@cdc.gov
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  C. A. Beamer and A. Holian Scavenger receptor class A type I/II (CD204) null mice fail to develop fibrosis following silica exposure Am J Physiol Lung Cell Mol Physiol, August 1, 2005; 289(2): L186 - L195. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. D. Taylor, J. R. Roberts, A. F. Hubbs, M. J. Reasor, and J. M. Antonini Quantitative Image Analysis of Drug-Induced Lung Fibrosis Using Laser Scanning Confocal Microscopy Toxicol. Sci., June 1, 2002; 67(2): 295 - 302. [Abstract] [Full Text] [PDF]
|
|