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Time-Dependent Resolution of Collagen Deposition During Skin Repair in Rats: A Correlative Morphological and Biochemical Study

Published online by Cambridge University Press:  05 November 2015

Rômulo D. Novaes ,
Marli C. Cupertino ,
Mariaurea M. Sarandy ,
André Souza ,
Evelise A. Soares  and
Reggiani V. Gonçalves
Rômulo D. Novaes*
Affiliation:
Department of Structural Biology, Federal University of Alfenas, Rua Gabriel Monteiro da Silva, 700, Campus Universitário, Centro, Alfenas, MG 37130-000, Brazil
Marli C. Cupertino
Affiliation:
Department of General Biology, Federal University of Viçosa, Viçosa, MG 36570-000, Brazil
Mariaurea M. Sarandy
Affiliation:
Department of General Biology, Federal University of Viçosa, Viçosa, MG 36570-000, Brazil
André Souza
Affiliation:
Department of Animal Biology, Federal University of Viçosa, Viçosa, MG 36570-000, Brazil
Evelise A. Soares
Affiliation:
Department of Anatomy, Federal University of Alfenas, Alfenas, MG 37130-000, Brazil
Reggiani V. Gonçalves
Affiliation:
Department of Animal Biology, Federal University of Viçosa, Viçosa, MG 36570-000, Brazil
*
*Corresponding author. romuonovaes@yahoo.com.br
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Abstract

Skin samples were used to compare microscopy methods used to quantify collagen with potential applicability to resolve time-dependent collagen deposition during skin wound healing in rats. Skin wounds by secondary intention were made in rats and tissue fragments were collected every 7 days for 21 days. Collagen content determined by biochemical analysis was compared with collagen measured by point counting (PC) on histological skin sections stained by Gomori’s trichrome method (Trichrome/PC), Sirius red under polarized light (PL) microscopy (Sirius red/PL-PC), and computational color segmentation (CS) applied to sections stained with Sirius red (Sirius red/PL-CS). All microscopy methods investigated resolved the time-dependent dynamics of collagen deposition in scar tissue during skin wound healing in rats. Collagen content measured by Sirius red/PL-PC and Sirius red/PL-CS was significantly lower when compared with Trichrome/PC. The Trichrome/PC method provided overestimated values of collagen compared with biochemical analysis. In the early stages of wound healing, which shows high production of noncollagenous molecules, Sirius red/PL-CS and Sirius red/PL-PC methods were more suitable for quantification of collagen fibers. Trichrome staining did not allow clear separation between collagenous and noncollagenous elements in skin samples, introducing a marked bias in collagen quantification.

Keywords

dermisextracellular matrixmorphologypathologywound healing
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 21 , Issue 6 , December 2015 , pp. 1482 - 1490
Copyright
© Microscopy Society of America 2015 

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