In Vitro Sirius Red Collagen Assay Measures the Pattern Shift from Soluble to Deposited Collagen

  • Chun Chen
  • Shanmin Yang
  • Mei Zhang
  • Zhenhuan Zhang
  • Bingrong Zhang
  • Deping Han
  • Jun Ma
  • Xiaohui Wang
  • Jingshen Hong
  • Yansong Guo
  • Paul Okunieff
  • Lurong Zhang
Conference paper
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 765)

Abstract

In this study, we compared two in vitro collagen production assays ([3H]-proline incorporation and Sirius Red) for their ability to determine the pattern shift from soluble to deposited collagen. The effect of the antifibrotic agent, triptolide (TPL), on collagen production was also studied. The results showed that: (1) 48 h after NIH 3T3 (murine embryo fibroblast) and HFL-1(human fetal lung fibroblast) were exposed to transforming growth factor-beta 1 (TGF-β), there was an increase in soluble collagen in the culture medium; (2) on day 4, soluble collagen declined, whereas deposited collagen increased; (3) Sirius Red was easier to use than [3H]-proline incorporation and more consistently reflected the collagen pattern shift from soluble to deposited; (4) the in vitro Sirius Red assay took less time than the in vivo assay to determine the effect of TPL. Our results suggest that: (a) the newly synthesized soluble collagen can sensitively evaluate an agent’s capacity for collagen production and (b) Sirius Red is more useful than [3H]-proline because it is easier to use, more convenient, less time consuming, and does not require radioactive material.

Keyword

Collagen 

Notes

Acknowledgments

This project is supported in part by U19 AI067733, RC1AI078519, RC2-AI-087580, RC1-AI081274 (NIAID/NIH), and Shands Cancer Center startup funds (University of Florida). We thank Kate Casey-Sawicki for editing this manuscript and Dr. Chihray Liu and his group for setting up the physical dosimetry for accurate radiation.

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Chun Chen
    • 1
  • Shanmin Yang
    • 1
  • Mei Zhang
    • 1
  • Zhenhuan Zhang
    • 1
  • Bingrong Zhang
    • 1
  • Deping Han
    • 1
  • Jun Ma
    • 1
  • Xiaohui Wang
    • 1
  • Jingshen Hong
    • 1
  • Yansong Guo
    • 1
  • Paul Okunieff
    • 1
  • Lurong Zhang
    • 1
  1. 1.Department of Radiation Oncology, UF Shands Cancer CenterUniversity of FloridaGainesvilleUSA

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