Quantitative analysis of collagen fiber organization in injured tendons using Fourier transform-second harmonic generation imaging.

Sivaguru, M
·
Durgam, S
·
Ambekar, R
·
Luedtke, D
·
Fried, G
·
Stewart, A
·
Toussaint, KC

Abstract: Fourier transform-second harmonic generation (FT-SHG) imaging is used as a technique for evaluating collagenase-induced injury in horse tendons. The differences in collagen fiber organization between normal and injured tendon are quantified. Results indicate that the organization of collagen fibers is regularly oriented in normal tendons and randomly organized in injured tendons. This is further supported through the use of additional metrics, in particular, the number of dark (no/minimal signal) and isotropic (no preferred fiber orientation) regions in the images, and the ratio of forward-to-backward second-harmonic intensity. FT-SHG microscopy is also compared with the conventional polarized light microscopy and is shown to be more sensitive to assessing injured tendons than the latter. Moreover, sample preparation artifacts that affect the quantitative evaluation of collagen fiber organization can be circumvented by using FT-SHG microscopy. The technique has potential as an assessment tool for evaluating the impact of various injuries that affect collagen fiber organization.

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Publication Date: 2010-12-18 PubMed ID: 21164843DOI: 10.1364/OE.18.024983Google Scholar: Lookup

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Non-P.H.S.

Summary

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The research paper examines the use of Fourier transform-second harmonic generation (FT-SHG) imaging in evaluating injury to horse tendons by assessing the organization of collagen fibers. The study determined that this technique notably displays the distinctions between normal tendons and those injured, with potential for assessing the impact of various injuries that affect collagen fiber organization.

Methodology

The study makes use of Fourier transform-second harmonic generation (FT-SHG) imaging, which is a technique for observing the microscopic features of biological tissues. In this specific investigation, it was used to evaluate the injury in horse tendons caused by collagenase.
The main focus was to ascertain the differences in the organization of collagen fibers between normal and injured tendons.
Extra metrics that were utilized included the number of dark regions (signifying no or minimal signal) and isotropic regions (implying no preferred fiber orientation) within the images. Additionally, the ratio of forward-to-backward second harmonic intensity was observed.

Results and Findings

From the results, the researchers deduced that normally, the collagen fibers in the tendons have regular, ordered arrangements. Contrarily, when injured, the fibers are organized randomly.
A comparison is provided between FT-SHG microscopy and conventional polarized light microscopy. FT-SHG microscopy was found to be more sensitive when it comes to assessing injured tendons.
Interestingly, FT-SHG microscopy was recognized to overcome limitations of sample preparation artifacts that might otherwise affect the quantitative evaluation of collagen fiber organization. This essentially bolsters the reliability of the findings obtained through its use.

Implications

The research displays the potential of FT-SHG microscopy as an effective tool for assessing injuries affecting collagen fiber organization.
This technique could enable more accurate determination of tendon damage and subsequently, the extent of injury in horses and possibly other animals or even humans.
The advancement of this technique could, therefore, hold significant implications for veterinary and medical science, particularly in the area of orthopedics.

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APA

Sivaguru M, Durgam S, Ambekar R, Luedtke D, Fried G, Stewart A, Toussaint KC. (2010). Quantitative analysis of collagen fiber organization in injured tendons using Fourier transform-second harmonic generation imaging. Opt Express, 18(24), 24983-24993. https://doi.org/10.1364/OE.18.024983

Publication

Optics express

ISSN: 1094-4087

NlmUniqueID: 101137103

Country: United States

Language: English

Volume: 18

Issue: 24

Pages: 24983-24993

Researcher Affiliations

Sivaguru, Mayandi

Institute for Genomic Biology, University of Illinois Urbana-Champaign, 1206 West Gregory Drive, Urbana, Illinois 61801, USA.

Durgam, Sushmitha

Ambekar, Raghu

Luedtke, David

Fried, Glenn

Stewart, Allison

Toussaint, Kimani C

MeSH Terms

Animals
Fibrillar Collagens / analysis
Fibrillar Collagens / chemistry
Fourier Analysis
Horses
Microscopy, Polarization / methods
Tendons / pathology

Citations

This article has been cited 46 times.

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