Determination of Fibrosis from Cryostat Sections Using High Performance Liquid Chromatography: Skeletal Muscle

Details

• Personal Author:
  Miller, G. Roger ; Smith CA ; Stauber WT
• Description:
  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. [Description provided by NIOSH]
• Subjects:
  Animals Chromatography Fibrosis Histology Laboratories Musculoskeletal Diseases Musculoskeletal System Occupational Health Safety
• Keywords:
  Animal Studies Chromatographic Analysis Collagen Fibrils Laboratory Animals Muscle Cells Muscle Tissue Musculoskeletal System Disorders
• ISSN:
  0018-2214
• Document Type:
  Text
• Funding:
  Grant
• Genre:
  Journal Article
• Place as Subject:
  OSHA Region 3 ; West Virginia
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Pages in Document:
  89-94
• Volume:
  31
• Issue:
  2
• NIOSHTIC Number:
  nn:20024469
• Citation:
  Histochem J 1999 Feb; 31(2):89-94
• Contact Point Address:
  Department of Physiology, West Virginia University, PO Box 9229, Morgantown, WV 26506, USA
• Federal Fiscal Year:
  1999
• NORA Priority Area:
  Disease and Injury: Musculoskeletal Disorders of the Upper Extremities
• Performing Organization:
  West Virginia University, Morgantown, West Virginia
• Peer Reviewed:
  True
• Start Date:
  19920101
• Source Full Name:
  The Histochemical Journal
• End Date:
  20050531
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:24d1e2b4257dc1dc7bcc9555b1cc9f3b28567210f6af50335cbe9b88b9b3afa9dface0498969bb69db678c94aed937b2788fe377b059b2d89476692428d83992
• Download URL:
  https://stacks.cdc.gov/view/cdc/196259/cdc_196259_DS1.pdf
• File Type:
  [PDF - 779.48 KB ]