A new strategy for the decellularisation of large equine tendons as biocompatible tendon substitutes.
Abstract: Tendon ruptures and/or large losses remain to be a great clinical challenge and often require full replacement of the damaged tissue. The use of auto- and allografts or engineered scaffolds is an established approach to restore severe tendon injuries. However, these grafts are commonly related to scarce biocompatibility, site morbidity, chronic inflammation and poor biomechanical properties. Recently, the decellularisation techniques of allo- or xenografts using specific detergents have been studied and have been found to generate biocompatible substitutes that resemble the native tissue. This study aims to identify a novel decellularisation protocol for large equine tendons that would produce an extracellular matrix scaffold suitable for the regeneration of injured tendons in humans. Specifically, equine tendons were treated either with tri (n-butyl) phosphate alone, or associated to multiple concentrations of peracetic acid (1, 3 and 5 %), which has never before been tested in vitro.Samples were then analysed by histology and with biochemical, biomechanical, and cytotoxicity tests. The best decellularisation protocol, resulting from these examinations, was selected and the chosen scaffold was re-seeded with murine fibroblasts. Resulting grafts were tested for cell viability, histologic analysis, DNA and collagen content. The results identified 1 % tri (n-butyl) phosphate combined with 3 % peracetic acid as the most suitable decellularised matrix in terms of biochemical and biomechanical properties. Moreover, the non-cytotoxic nature of the decellularised matrix allowed for good fibroblast reseeding, thus demonstrating a biocompatible matrix that will be suitable for tendon tissue engineering and hopefully as substitutes in severe tendon damages.
Publication Date: 2016-07-08 PubMed ID: 27386840DOI: 10.22203/ecm.v032a04Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article presents a new approach for decellularising large horse tendons to create biocompatible tendon replacements. The study established a specific decellularisation protocol that produced a scaffold from these tendons that could potentially aid in the regeneration of injured human tendons.
Objective of the Research
- The main objective of the research is to establish a novel decellularisation protocol for large equine (horse) tendons. The goal is to create an extracellular matrix scaffold that could assist in regenerating injured tendons in humans.
Research Methodology
- To accomplish this, equine tendons were subjected to specific treatments. They were treated with tri (n-butyl) phosphate alone or with various concentrations of peracetic acid (1, 3 and 5% ), an approach yet to be tested in vitro.
- The treated samples were then analysed through various tests, including histology, biochemical, biomechanical, and cytotoxicity tests.
Findings from the Study
- The results identified that 1% tri (n-butyl) phosphate combined with 3% peracetic acid yielded the most suitable decellularised matrix in terms of biochemical and biomechanical properties.
- Importantly, it was also established that the decellularised matrix was not cytotoxic (harmful to cells), which allowed for successful reseeding with murine (mouse) fibroblasts.
Conclusion and Implication
- The study concludes that the established decellularisation protocol yields a biocompatible matrix that is suitable for tendon tissue engineering.
- Furthermore, this matrix could potentially serve as a substitute in cases of severe tendon damage in humans, thereby addressing a significant clinical challenge.
Cite This Article
APA
Bottagisio M, Pellegata AF, Boschetti F, Ferroni M, Moretti M, Lovati AB.
(2016).
A new strategy for the decellularisation of large equine tendons as biocompatible tendon substitutes.
Eur Cell Mater, 32, 58-73.
https://doi.org/10.22203/ecm.v032a04 Publication
Researcher Affiliations
- Cell and Tissue Engineering Laboratory, IRCCS Galeazzi Orthopaedic Institute, via R. Galeazzi 4, 20161 Milan, Italy.arianna.lovati@grupposandonato.it.
MeSH Terms
- Animals
- Biocompatible Materials / pharmacology
- Biomechanical Phenomena
- Cell Death / drug effects
- Collagen / metabolism
- DNA / metabolism
- Elastic Modulus
- Extracellular Matrix / metabolism
- Glycosaminoglycans / metabolism
- Horses
- Stress, Mechanical
- Sulfates / metabolism
- Tendons / cytology
- Tissue Engineering / methods
Citations
This article has been cited 9 times.- El Soury M, García-García ÓD, Moretti M, Perroteau I, Raimondo S, Lovati AB, Carriel V. Comparison of Decellularization Protocols to Generate Peripheral Nerve Grafts: A Study on Rat Sciatic Nerves. Int J Mol Sci 2021 Feb 27;22(5).
- Sun Y, Lovric V, Wang T, Oliver RA, Walsh WR. Effects of SCCO(2), Gamma Irradiation, and Sodium Dodecyl Sulfate Treatments on the Initial Properties of Tendon Allografts. Int J Mol Sci 2020 Feb 25;21(5).
- Bottagisio M, D'Arrigo D, Talò G, Bongio M, Ferroni M, Boschetti F, Moretti M, Lovati AB. Achilles Tendon Repair by Decellularized and Engineered Xenografts in a Rabbit Model. Stem Cells Int 2019;2019:5267479.
- Talò G, D'Arrigo D, Lorenzi S, Moretti M, Lovati AB. Independent, Controllable Stretch-Perfusion Bioreactor Chambers to Functionalize Cell-Seeded Decellularized Tendons. Ann Biomed Eng 2020 Mar;48(3):1112-1126.
- Lovati AB, D'Arrigo D, Odella S, Tos P, Geuna S, Raimondo S. Nerve Repair Using Decellularized Nerve Grafts in Rat Models. A Review of the Literature. Front Cell Neurosci 2018;12:427.
- Liu J, Saul D, Böker KO, Ernst J, Lehman W, Schilling AF. Current Methods for Skeletal Muscle Tissue Repair and Regeneration. Biomed Res Int 2018;2018:1984879.
- Roth SP, Glauche SM, Plenge A, Erbe I, Heller S, Burk J. Automated freeze-thaw cycles for decellularization of tendon tissue - a pilot study. BMC Biotechnol 2017 Feb 14;17(1):13.
- Tavares DF, Mano JF, Oliveira MB. Advances in abiotic tissue-based biomaterials: A focus on decellularization and devitalization techniques. Mater Today Bio 2025 Jun;32:101735.
- Kouroupis D, Perucca Orfei C, Correa D, Talò G, Libonati F, De Luca P, Raffo V, Best TM, de Girolamo L. Cellular and Structural Changes in Achilles and Patellar Tendinopathies: A Pilot In Vivo Study. Biomedicines 2024 Apr 30;12(5).
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