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PloS one2013; 8(5); e64151; doi: 10.1371/journal.pone.0064151

Functional characterization of detergent-decellularized equine tendon extracellular matrix for tissue engineering applications.

Abstract: Natural extracellular matrix provides a number of distinct advantages for engineering replacement orthopedic tissue due to its intrinsic functional properties. The goal of this study was to optimize a biologically derived scaffold for tendon tissue engineering using equine flexor digitorum superficialis tendons. We investigated changes in scaffold composition and ultrastructure in response to several mechanical, detergent and enzymatic decellularization protocols using microscopic techniques and a panel of biochemical assays to evaluate total protein, collagen, glycosaminoglycan, and deoxyribonucleic acid content. Biocompatibility was also assessed with static mesenchymal stem cell (MSC) culture. Implementation of a combination of freeze/thaw cycles, incubation in 2% sodium dodecyl sulfate (SDS), trypsinization, treatment with DNase-I, and ethanol sterilization produced a non-cytotoxic biomaterial free of appreciable residual cellular debris with no significant modification of biomechanical properties. These decellularized tendon scaffolds (DTS) are suitable for complex tissue engineering applications, as they provide a clean slate for cell culture while maintaining native three-dimensional architecture.
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Publication Date: 2013-05-27 PubMed ID: 23724028PubMed Central: PMC3664617DOI: 10.1371/journal.pone.0064151Google Scholar: Lookup
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  • Journal Article
  • Research Support
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  • Extramural
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  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article is about the process of creating an effective biological scaffold using horse tendons for use in tissue engineering for orthopedic purposes.

Background

Natural extracellular matrix (ECM) offers unique advantages for orthopedic tissue engineering, owing to its inherent functional properties. ECM components have a high potential for biocompatibility, the ability to direct native cell behavior, and intriguing possibilities for the repair and regeneration of various tissue types, including tendons.

In the study, the researchers aimed to create an optimal biological scaffold using equine flexor digitorum superficialis tendons (part of a horse’s leg structure).

Process Overview

  • The research team explored changes in the structure and composition of the scaffold in relation to various decellularization methods. These methods involved mechanical, detergent, and enzymatic protocols.
  • Techniques like microscopy and biochemical assays were employed to assess the total protein, collagen, glycosaminoglycan, and DNA levels before and after the decellularization process.
  • The team also tested the biocompatibility of the treated ECM through static mesenchymal stem cell (MSC) culture to verify its potential use as an environment promoting cell growth.

Findings

According to the study:

  • A combination of several processes – freezing and thawing, exposure to 2% sodium dodecyl sulfate (SDS), trypsin treatment, DNase-I treatment, and sterilization with ethanol, resulted in a biologically suitable material that was free from significant cellular debris and was not cytotoxic (not harmful to cells).
  • The treated biomaterial did not undergo noticeable alterations to its biomechanical properties, making it ideal for sophisticated tissue engineering applications.
  • The derived decellularized tendon scaffold (DTS) retained the native three-dimensional architecture crucial for effective tissue engineering.

In conclusion, the research presents extensive testing and validation of a novel protocol for tendon scaffold preparation, ultimately leading to a ‘clean slate’ for cell culture. The absence of cytotoxic effects and preserved biomechanical properties of the treated ECM makes it suitable for advanced tissue engineering applications such as orthopedic tissue replacements.

Cite This Article

APA
Youngstrom DW, Barrett JG, Jose RR, Kaplan DL. (2013). Functional characterization of detergent-decellularized equine tendon extracellular matrix for tissue engineering applications. PLoS One, 8(5), e64151. https://doi.org/10.1371/journal.pone.0064151

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 8
Issue: 5
Pages: e64151
PII: e64151

Researcher Affiliations

Youngstrom, Daniel W
  • Department of Biomedical and Veterinary Sciences, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Tech, Leesburg, Virginia, United States of America.
Barrett, Jennifer G
    Jose, Rod R
      Kaplan, David L

        MeSH Terms

        • Animals
        • Biocompatible Materials / chemistry
        • Cell Culture Techniques
        • Cell Proliferation
        • Detergents / chemistry
        • Extracellular Matrix / chemistry
        • Extracellular Matrix / ultrastructure
        • Glycosaminoglycans / chemistry
        • Horses
        • Male
        • Mesenchymal Stem Cells / cytology
        • Tendons / chemistry
        • Tendons / ultrastructure
        • Tissue Engineering
        • Tissue Scaffolds / chemistry

        Grant Funding

        • P41 EB002520 / NIBIB NIH HHS

        Conflict of Interest Statement

        The authors have declared that no competing interests exist.

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