Using a freeze substitution fixation technique and histological crimp analysis for characterizing regions of strain in ligaments loaded in situ

Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society
Richard S BoormanJohn M Clark

Abstract

Type I collagen fibrils in tendons and ligaments assume a sinusoidal wave shape, or crimp, which straightens only with tensile load. The load response of crimp has been studied primarily in isolated subunits and not in complex, intact structures. The purpose of our study was to determine if freeze substitution fixation of an entire ligament could preserve changes in crimp morphology induced by functionally relevant loading conditions. We hypothesized that, in ligaments prepared by freeze-substitution fixation under load, crimp would progressively extinguish with increasing loads, and nonuniform strain following partial section could be detected from crimp morphology. Tensile loads ranging from 0 to 220 N were applied to patellar ligaments of 16 fresh rabbit stifle joints using simulated isometric quadriceps pull through the patella. The loaded joints were flash frozen with isopentane cooled in liquid nitrogen, then fixed using freeze substitution. Another six ligaments were loaded to 150 N following incision of the anterior third and evaluated under polarized light microscopy for crimp distribution. Ligaments with no or low loads could be identified by the presence of crimp on mid-sagittal sections. Strain distribution was inho...Continue Reading

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Citations

Nov 14, 2007·Annals of Biomedical Engineering·Diego F VillegasTammy L Haut Donahue
May 18, 2013·Bulletin of Mathematical Biology·Mark S Thompson
Dec 17, 2008·Journal of Applied Physiology·Trevor J LujanJeffrey A Weiss
Nov 24, 2016·Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society·Michael LavagninoSteven P Arnoczky
Jun 23, 2009·Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society·Spencer P LakeLouis J Soslowsky
Jul 9, 2013·Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society·Kazuya IkomaToshikazu Kubo
Oct 17, 2013·Journal of Anatomy·Carlos I Lorda-DiezJuan A Montero
Feb 14, 2015·Journal of Anatomy·Lei ZhaoNeil Broom
Apr 17, 2018·Experimental Eye Research·Ning-Jiun JanIan A Sigal

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