Summary
The structure of the collagen fibers that composes tendon and ligament are disrupted or damaged during injury and healing. Quantification of these changes is traditionally a laborious and subjective task. In this work we apply two automated techniques, Fourier transformation (FFT) and fractal dimension analysis (FA) to quantify the organization of collagen fibrils. Using multi-photon images we show that for healing ligament FA differentiates more clearly between the different time-points during healing. Using scanning electron microcopy images of overstretched tendon we show that combining FFT and FA measures separates the damaged and undamaged groups more clearly than either method individually.
The mechanical behavior of tendons has been well studied in vitro. A noninvasive method to acquire mechanical data would be highly beneficial. Elastography has been a promising method of gathering in vivo tissue mechanical behavior, but it has inherent limitations. This study presents acoustoelasticity as an alternative ultrasound-based method of measuring tendon stress and strain by reporting a relationship between ultrasonic echo intensity (B mode ultrasound image brightness) and mechanical behavior of tendon in vitro. Porcine digital flexor tendons were cyclically loaded in a mechanical testing system while ultrasonic echo response was recorded. We report that echo intensity closely follows the applied cyclic strain pattern in time with higher strain protocols resulting in larger echo intensity changes. We also report that echo intensity is related nonlinearly to stress and nearly linearly to strain. This indicates that ultrasonic echo intensity is related to the mechanical behavior in a loaded tissue by an acoustoelastic response, as previously described in homogeneous, nearly incompressible materials.
Acoustoelasticity is therefore able to relate strain-dependent stiffness and stress to the reflected echo, even in the processed B-mode signals reflected from viscoelastic, inhomogeneous material such as tendon, and is a promising metric to acquire in vivo mechanical data noninvasively.
Ligament healing of a grade III injury (i.e. a complete tear) involves a multifaceted chain of events that forms a neoligament, which is more scar-like in character than the native tissue. The remodeling process may last months or even years with the injured ligament never fully recovering pre-injury mechanical properties. With tissue engineering and regenerative medicine, understanding the normal healing process in ligament and quantifying it provide a basis to create and assess innovative treatments. Ligament fibroblasts produce a number of ECM components, including collagen types I and III, decorin and fibromodulin. Using a combination of advanced histology, molecular biology and nonlinear optical imaging approaches, the early ECM events during ligament healing have been better characterized and defined. First, the dynamic changes in ECM factors after injury are shown. Second, the factors associated with creeping substitution are identified. Finally, a method to quantify collagen organization is developed and used. Each ECM factor described herein as well as the temporal quantification of fiber organization helps elucidate the complexity of ligament healing.
Cervical arthroplasty should be further investigated in vivo to determine if it is a viable alternative to the ventral slot or pin-PMMA procedures for surgical treatment of cervical diseases in dogs and in particular for treatment of disk-associated caudal cervical spondylomyelopathy.
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