Skip directly to search Skip directly to A to Z list Skip directly to page options Skip directly to site content

NIOSHTIC-2 Publications Search

Search Results

Determination of fibrosis from cryostat sections using high performance liquid chromatography: skeletal muscle.

Authors
Miller-GR; Smith-CA; Stauber-WT
Source
Histochem J 1999 Feb; 31(2):89-94
NIOSHTIC No.
20024469
Abstract
Analysis of hydroxyproline (collagen) and pyridinoline (collagen cross-links) in biopsies prepared for routine histological evaluation with OCT compound was performed. Frozen sections (250 microm-thick) were cut from cardiac muscle, diaphragm, liver, and soleus muscle from the rat. After removal of OCT compound by rinsing, the samples were dried, weighed and hydrolyzed in 6 N HCl. A portion of the hydrolysate was analyzed for hydroxyproline using high performance liquid chromatography with collagen type I as the standard. Collagen concentrations ranged from 6.6 microg/mg dry weight (liver) to 74.7 microg/mg dry weight (diaphragm). From the remainder of the hydrolyzate, pyridinoline cross-links of collagen were separated and analyzed similarly by high performance liquid chromatography. The concentration of pyridinoline ranged from 2.6 ng/mg dry weight (liver) to 35.6 ng/mg dry weight (diaphragm). These techniques were adequate to analyze both collagen and pyridinoline (i.e. collagen cross-links) in small biopsy samples (< 1 mg dry weight) routinely used in clinical pathology. The method proved useful in the quantitation of focal fibrosis in a partially denervated rat soleus. Denervation was confirmed using fast myosin immunohistochemistry which revealed large areas of small myofibres containing fast myosin. Collagen concentration increased by five-fold and collagen cross-links by more than 7-fold consistent with fibrotic changes known to occur with denervation.
Keywords
Fibrosis; Chromatographic-analysis; Collagen-fibrils; Histology; Muscle-tissue; Muscle-cells; Musculoskeletal-system-disorders; Laboratory-animals; Animals; Animal-studies
Contact
Department of Physiology, West Virginia University, PO Box 9229, Morgantown, WV 26506, USA
CODEN
HISJAE
Publication Date
19990201
Document Type
Journal Article
Funding Amount
868663
Funding Type
Grant
Fiscal Year
1999
NTIS Accession No.
NTIS Price
Identifying No.
Grant-Number-R01-OH-002918
Issue of Publication
2
ISSN
0018-2214
Priority Area
Disease and Injury: Musculoskeletal Disorders of the Upper Extremities
Source Name
The Histochemical Journal
State
WV
Performing Organization
West Virginia University, Morgantown, West Virginia
TOP