Niels Hammer1, Daniel Huster2, Andreas Boldt3, Carsten Hädrich4, Holger Koch5, Robert Möbius6, Gundula Schulze-Tanzil7, Holger A Scheidt2. 1. Department of Anatomy, University of Otago, 270 Great King Street, Dunedin 9054, New Zealand. Electronic address: nlshammer@googlemail.com. 2. Institute of Medical Physics and Biophysics, University of Leipzig, Faculty of Medicine, Leipzig, Germany. 3. Institute of Clinical Immunology, University of Leipzig, Faculty of Medicine, Leipzig, Germany; Translational Centre for Regenerative Medicine, University of Leipzig, Faculty of Medicine, Leipzig, Germany. 4. Institute of Forensic Medicine, University of Leipzig, Faculty of Medicine, Leipzig, Germany. 5. Translational Centre for Regenerative Medicine, University of Leipzig, Faculty of Medicine, Leipzig, Germany. 6. Institute of Anatomy, University of Leipzig, Faculty of Medicine, Leipzig, Germany. 7. Department of Anatomy, Nuremberg Hospital Medical School, Paracelsus Medical University, Nuremburg, Germany and Salzburg, Austria.
Abstract
INTRODUCTION: Acellular scaffolds are frequently used for the surgical repair of ligaments and tendons. Even though data on the macro-mechanical properties related to the acellularization process exist, corresponding data on the nano-structural properties are still lacking. Such data would help identify target proteins of the formed extracellular matrix that are chemically altered by the acellularization. In this study we examined the altered structure by comparing molecular properties of collagens from native and acellular iliotibial tract samples to the macroscopic stress-strain behavior of tract samples. MATERIAL AND METHODS: Matched pairs of five human iliotibial tract samples were obtained from five donors (mean age 28.2±4.7 years). One of each pair was acellularized using 1vol% sodium dodecyl sulfate (SDS) for 7 days. (13)C magic angle spinning nuclear magnetic resonance spectroscopy ((13)C CP MAS NMR) was utilized to compare the collagen overall secondary structure and internal dynamics of collagen-typical amino acid proteins. The resulting data was compared to age-matched stress-strain data of tract samples obtained in an uniaxial tensile setup and histologically. RESULTS: Typical and nearly identical collagen (13)C CP MAS NMR spectra were found in the tract samples before and after acellularization with SDS. The characteristic collagen backbone remained intact in the native and acellular samples. Collagen molecular composition was largely unaltered in both conditions. Furthermore, a similar dynamic behavior was found for the amino acids Hyp γ, Pro α/Hyp α, Ala α, Gly α and Ala β. These minute alterations in the collagens' molecular properties related to acellularization with SDS were in line with the similarly minute changes in the macro-mechanical tensile behavior, such as the elastic modulus and ultimate stress. Histology showed intact type I collagens, minute amounts of elastins before and after acellularization and evidence for acellularization-induced reductions of proteoglycans. DISCUSSION: Nano-structural properties of collagens are minutely affected by SDS treatment for acellularization, indicated by the molecular composition and dynamics. The lacking acellularization-related changes in the molecular structure properties of collagens in iliotibial tract samples are in line with the small alterations in their macro-mechanical tensile behavior. Though the given setup approaches soft tissue mechanics from both scaling extremes of mechanical testing, further structural analyzes are needed in a larger sample size to substantiate these findings.
INTRODUCTION: Acellular scaffolds are frequently used for the surgical repair of ligaments and tendons. Even though data on the macro-mechanical properties related to the acellularization process exist, corresponding data on the nano-structural properties are still lacking. Such data would help identify target proteins of the formed extracellular matrix that are chemically altered by the acellularization. In this study we examined the altered structure by comparing molecular properties of collagens from native and acellular iliotibial tract samples to the macroscopic stress-strain behavior of tract samples. MATERIAL AND METHODS: Matched pairs of five human iliotibial tract samples were obtained from five donors (mean age 28.2±4.7 years). One of each pair was acellularized using 1vol% sodium dodecyl sulfate (SDS) for 7 days. (13)C magic angle spinning nuclear magnetic resonance spectroscopy ((13)C CP MAS NMR) was utilized to compare the collagen overall secondary structure and internal dynamics of collagen-typical amino acid proteins. The resulting data was compared to age-matched stress-strain data of tract samples obtained in an uniaxial tensile setup and histologically. RESULTS: Typical and nearly identical collagen (13)C CP MAS NMR spectra were found in the tract samples before and after acellularization with SDS. The characteristic collagen backbone remained intact in the native and acellular samples. Collagen molecular composition was largely unaltered in both conditions. Furthermore, a similar dynamic behavior was found for the amino acids Hyp γ, Pro α/Hyp α, Ala α, Gly α and Ala β. These minute alterations in the collagens' molecular properties related to acellularization with SDS were in line with the similarly minute changes in the macro-mechanical tensile behavior, such as the elastic modulus and ultimate stress. Histology showed intact type I collagens, minute amounts of elastins before and after acellularization and evidence for acellularization-induced reductions of proteoglycans. DISCUSSION: Nano-structural properties of collagens are minutely affected by SDS treatment for acellularization, indicated by the molecular composition and dynamics. The lacking acellularization-related changes in the molecular structure properties of collagens in iliotibial tract samples are in line with the small alterations in their macro-mechanical tensile behavior. Though the given setup approaches soft tissue mechanics from both scaling extremes of mechanical testing, further structural analyzes are needed in a larger sample size to substantiate these findings.
Authors: Stefan Schleifenbaum; Torsten Prietzel; Gabriela Aust; Andreas Boldt; Sebastian Fritsch; Isabel Keil; Holger Koch; Robert Möbius; Holger A Scheidt; Martin F X Wagner; Niels Hammer Journal: PLoS One Date: 2016-03-09 Impact factor: 3.240