title: Micro-mechanical properties of the tendon-to-bone attachment.
abstract: The tendon-to-bone attachment (enthesis) is a complex hierarchical tissue that connects stiff bone to compliant tendon. The attachment site at the micrometer scale exhibits gradients in mineral content and collagen orientation, which likely act to minimize stress concentrations. The physiological micromechanics of the attachment thus define resultant performance, but difficulties in sample preparation and mechanical testing at this scale have restricted understanding of structure-mechanical function. Here, microscale beams from entheses of wild type mice and mice with mineral defects were prepared using cryo-focused ion beam milling and pulled to failure using a modified atomic force microscopy system. Micromechanical behavior of tendon-to-bone structures, including elastic modulus, strength, resilience, and toughness, were obtained. Results demonstrated considerably higher mechanical performance at the micrometer length scale compared to the millimeter tissue length scale, describing enthesis material properties without the influence of higher order structural effects such as defects. Micromechanical investigation revealed a decrease in strength in entheses with mineral defects. To further examine structure-mechanical function relationships, local deformation behavior along the tendon-to-bone attachment was determined using local image correlation. A high compliance zone near the mineralized gradient of the attachment was clearly identified and highlighted the lack of correlation between mineral distribution and strain on the low-mineral end of the attachment. This compliant region is proposed to act as an energy absorbing component, limiting catastrophic failure within the tendon-to-bone attachment through higher local deformation. This understanding of tendon-to-bone micromechanics demonstrates the critical role of micrometer scale features in the mechanics of the tissue.
  
  
  
  The tendon-to-bone attachment (enthesis) is a complex hierarchical tissue with features at a numerous scales that dissipate stress concentrations between compliant tendon and stiff bone. At the micrometer scale, the enthesis exhibits gradients in collagen and mineral composition and organization. However, the physiological mechanics of the enthesis at this scale remained unknown due to difficulty in preparing and testing micrometer scale samples. This study is the first to measure the tensile mechanical properties of the enthesis at the micrometer scale. Results demonstrated considerably enhanced mechanical performance at the micrometer length scale compared to the millimeter tissue length scale and identified a high-compliance zone near the mineralized gradient of the attachment. This understanding of tendon-to-bone micromechanics demonstrates the critical role of micrometer scale features in the mechanics of the tissue.
authors.affiliation: Dept. of Orthopedic Surgery, Columbia University, New York, NY, United States. Electronic address: alix.c.deymier@gmail.com.
authors.forename: Alix C
authors.initials: AC
authors.lastname: Deymier
authors.affiliation: School of Engineering and Materials Science, Queen Mary University of London, UK. Electronic address: yiran.an@auckland.ac.nz.
authors.forename: Yiran
authors.initials: Y
authors.lastname: An
authors.affiliation: Dept. of Biomedical Engineering, Washington University, St Louis, MO, United States. Electronic address: jb3911@columbia.edu.
authors.forename: John J
authors.initials: JJ
authors.lastname: Boyle
authors.affiliation: Dept. of Orthopaedic Surgery, Washington University, St Louis, MO, United States. Electronic address: annie.gitomer@wustl.edu.
authors.forename: Andrea G
authors.initials: AG
authors.lastname: Schwartz
authors.affiliation: Missouri S&T Global - St. Louis and Dept. of Mechanical and Aerospace Engineering, Missouri University of Science and Technology, St. Louis, MO, United States. Electronic address: vbirman@mst.edu.
authors.forename: Victor
authors.initials: V
authors.lastname: Birman
authors.affiliation: Dept. of Mechanical Engineering and Materials Science, Washington University, St Louis, MO, United States. Electronic address: genin@wustl.edu.
authors.forename: Guy M
authors.initials: GM
authors.lastname: Genin
authors.affiliation: Dept. of Orthopedic Surgery, Columbia University, New York, NY, United States. Electronic address: sat2@columbia.edu.
authors.forename: Stavros
authors.initials: S
authors.lastname: Thomopoulos
authors.affiliation: School of Engineering, University of Portsmouth, Portsmouth, UK. Electronic address: asa.barber@port.ac.uk.
authors.forename: Asa H
authors.initials: AH
authors.lastname: Barber
mesh_terms: D000818:Animals; D015519:Bone Density; D055119:Elastic Modulus; D005260:Female; D058430:Humeral Head / Q000737:chemistry*; D051379:Mice; D013710:Tendons / Q000737:chemistry*
keywords: AFM; Enthesis; Gradient; Image correlation; Micromechanics; Tensile testing