Influence of sensor size on the accuracy of in-vivo ligament and tendon force measurements.

In-vivo tendon forces are commonly measured using transducers, which detect tension in the tendon fibers. A poorly understood source of measurement errors is the difference in stress distribution within the tendon between experimental and transducer calibration conditions. The objective of this study was to investigate this source of error, and to determine whether these errors could be minimized by proper selection of transducer size. The study was conducted using the infrapatellar ligament (patellar tendon) of New Zealand White rabbits. Tendon force was measured with two different size implantable force transducers (IFTs), one Wide and one Narrow, and by a strain gaged load cell in series with the tendon. Tests were conducted at five different loading conditions selected to produce five different stress distributions within the tendon. One loading condition corresponded to a typical post-experiment calibration, and the data from that condition were used to develop a calibration equation for the transducer. The errors that resulted from using this calibration were determined by comparing the tendon force measured by the in-series load cell with the force predicted from the IFT output using the calibration equation. Changes in stress distribution produced measurement errors up to 64 N with the Narrow IFT but only 24 N with the Wide IFT. We found the measurement error was dependent on sensor width. Our results support the hypothesis that measurement errors can be caused by differences in tendon stress distribution between calibration and experimental conditions. We further showed that these errors can be minimized by using an IFT, which samples the tension in a large percentage of the tendon fibers. Information from this study can be used for selection of an appropriately sized implantable force transducer for measuring tendon and ligament force.     

Simultaneous measurement of three-dimensional joint kinematics and ligament strains with optical methods

The objective of this study was to assess the precision and accuracy of a nonproprietary, optical three-dimensional (3D) motion analysis system for the simultaneous measurement of soft tissue strains and joint kinematics. The system consisted of two high-resolution digital cameras and software for calculating the 3D coordinates of contrast markers. System precision was assessed by examining the variation in the coordinates of static markers over time. Three-dimensional strain measurement accuracy was assessed by moving contrast markers fixed distances in the field of view and calculating the error in predicted strain. Three-dimensional accuracy for kinematic measurements was assessed by simulating the measurements that are required for recording knee kinematics. The field of view (190 mm) was chosen to allow simultaneous recording of markers for soft tissue strain measurement and knee joint kinematics. Average system precision was between +/-0.004 mm and +/-0.035 mm, depending on marker size and camera angle. Absolute error in strain measurement varied from a minimum of +/-0.025% to a maximum of +/-0.142%, depending on the angle between cameras and the direction of strain with respect to the camera axes. Kinematic accuracy for translations was between +/-0.008 mm and +/-0.034 mm, while rotational accuracy was +/-0.082 deg to +/-0.160 deg. These results demonstrate that simultaneous optical measurement of 3D soft tissue strain and 3D joint kinematics can be performed while achieving excellent accuracy for both sets of measurements.     

Upper blepharoplasty with bony anatomical landmarks to avoid injury to trochlea and superior oblique muscle tendon with fat resection.

The trochlea and superior oblique muscle tendon separate the medial and central fat compartments in the upper lid. The purpose of this study was to determine anatomical landmarks to predict the location of and avoid injuring the trochlea and superior oblique muscle tendon with orbital fat resection during upper blepharoplasty. The trochlea and superior oblique muscle tendon were identified in 14 cadaver heads. Bony anatomical landmarks were identified to predict the oblique vector along which the trochlea and superior oblique tendon lie. The trochlea was measured in millimeters from the palpable superior orbital foramen. The oblique course of the superior oblique muscle tendon was measured from its medial location in the lateral direction in millimeters from the frontozygomatic suture. These measurements were obtained with 4.0-power loupe magnification. The trochlea was identified 10.0 +/- 0.9 mm inferior to the palpable superior orbital foramen. The superior oblique muscle tendon coursed laterally along an oblique vector to within 1 mm of the frontozygomatic suture for all 14 dissections. The vertical vector of the superior orbital foramen was measured 15.9 +/- 1.1 mm lateral to the medial canthus. The width of the bony orbit measured 42.2 +/- 1.6 mm. In two dissections, the superior orbital foramen could not be palpated, and the latter measurements were used to predict the superior orbital foramen. This anatomical study showed that when performing orbital fat resection with upper blepharoplasty, the trochlea and superior oblique muscle tendon can be identified and avoided with the above-described bony landmarks.     

An algorithm for automated analysis of ultrasound images to measure tendon excursion in vivo

The accuracy of an algorithm for the automated tracking of tendon excursion from ultrasound images was tested in three experiments. Because the automated method could not be tested against direct measurements of tendon excursion in vivo, an indirect validation procedure was employed. In one experiment, a wire "phantom" was moved a known distance across the ultrasound probe and the automated tracking results were compared with the known distance. The excursion of the musculotendinous junction of the gastrocnemius during frontal and sagittal plane movement of the ankle was assessed in a single cadaver specimen both by manual tracking and with a cable extensometer sutured to the gastrocnemius muscle. A third experiment involved estimation of Achilles tendon excursion in vivo with both manual and automated tracking. Root mean squared (RMS) error was calculated between pairs of measurements after each test. Mean RMS errors of less than 1 mm were observed for the phantom experiments. For the in vitro experiment, mean RMS errors of 8-9% of the total tendon excursion were observed. Mean RMS errors of 6-8% of the total tendon excursion were found in vivo. The results indicate that the proposed algorithm accurately tracks Achilles tendon excursion, but further testing is necessary to determine its general applicability.     

A noncontact,nondestructive method for quantifying intratissue deformations and strains

The function of soft connective tissues is frequently characterized by quantifying tissue strain (e.g., during joint motion). Conventional techniques for quantifying tendon and ligament strain typically provide surface measures, using markers, stain lines or instrumentation that may influence the tissue. An alternative approach is to quantify intratendinous strain by applying texture correlation analysis to magnetic resonance (MR) images. This paper reports the accuracy and reproducibility of this approach by (1) assessing the reproducibility of MR images, (2) assessing texture correlation accuracy using simulated displacements, and (3) comparing texture correlation measures of displacement and strain from MR images to conventional techniques.     

Full field strain measurements of collagenous tissue by tracking fiber alignment through vector correlation

Full field strain measurements of biological tissue during loading are often limited to the quantification of fiduciary marker displacements on the tissue surface. These marker measurements can lack the necessary spatial resolution to characterize non-uniform deformation and may not represent the deformation of the load-bearing collagen microstructure. To overcome these potential limitations, a method was developed to track the deformation of the collagen fiber microstructure in ligament tissue. Using quantitative polarized light imaging, fiber alignment maps incorporating both direction and alignment strength at each pixel were generated during facet capsular ligament loading. A grid of virtual markers was superimposed over the tissue in the alignment maps, and the maximization of a vector correlation calculation between fiber alignment maps was used to track marker displacement. Tracking error was quantified through comparisons to the displacements of excised ligament tissue (n=3); separate studies applied uniaxial tension to isolated facet capsular ligament tissue (n=4) to evaluate tracking capabilities during large tissue deformations. The average difference between virtual marker and tissue displacements was 0.07±0.06 pixels. This error in marker location produced principal strain measurements of 1.2±1.6% when markers were spaced 4 pixels apart. During tensile tissue loading, substantial inhomogeneity was detected in the strain field using vector correlation tracking, and the location of maximum strain differed from that produced by standard tracking techniques using coarser meshes. These findings provide a method to directly measure fiber network strains using quantitative fiber alignment data, enabling a better understanding of structure–function relationships in tissues at different length scales.     

How cyclic loading affects the migration of radio-opaque markers attached to tendon grafts using a new method: a study using roentgen stereophotogrammetric analysis (RSA)

An increase in anterior laxity following reconstruction of the anterior cruciate ligament (ACL) can result from lengthening of the graft construct in either the regions of fixation and/or the region of the graft substance between the fixations. RSA could be a useful technique to determine lengthening in these regions if a method can be devised for attaching radio-opaque markers to soft tissue grafts so that marker migration from repeated loading of the graft is limited. Therefore, the objectives of this study were 1) to develop a method for attaching radio-opaque markers to an ACL graft that limits marker migration within the graft, 2) to characterize the error of an RSA system used to study migration, and 3) to determine the maximum amount of migration and the time when it occurs during cyclic loading of ACL grafts. Tendon markers were constructed from a 0.8-mm tantalum ball and a stainless steel suture. Ten double-looped tendon grafts were passed through tibial tunnels drilled in bovine tibias and fixed with a tibial fixation device. Two tendon markers were sewn to one tendon bundle of each graft and the grafts were cyclically loaded for 225,000 cycles from 20 N to 170 N. At specified intervals, simultaneous radiographs were obtained of the tendon markers and a radiographic standard of known length. The bias and imprecision in measuring the length of the radiographic standard were 0.0 and 0.046 mm respectively. Marker migration was computed as the change in distance between the two tendon markers along the axis of the tibial tunnel. Marker migration was greatest after 225,000 cycles with a root mean square (RMS) value of less than 0.2 mm. Because the RMS value indicates the error introduced into measurements of lengthening and because this error is small, the method described for attaching markers to an ACL graft has the potential to be useful for determining lengthening of ACL graft constructs in in vivo studies in humans.     

A novel validation and calibration method for motion capture systems based on micro-triangulation

Motion capture systems are widely used to measure human kinematics. Nevertheless, users must consider system errors when evaluating their results. Most validation techniques for these systems are based on relative distance and displacement measurements. In contrast, our study aimed to analyse the absolute volume accuracy of optical motion capture systems by means of engineering surveying reference measurement of the marker coordinates (uncertainty: 0.75 mm). The method is exemplified on an 18 camera OptiTrack Flex13 motion capture system. The absolute accuracy was defined by the root mean square error (RMSE) between the coordinates measured by the camera system and by engineering surveying (micro-triangulation). The original RMSE of 1.82 mm due to scaling error was managed to be reduced to 0.77 mm while the correlation of errors to their distance from the origin reduced from 0.855 to 0.209. A simply feasible but less accurate absolute accuracy compensation method using tape measure on large distances was also tested, which resulted in similar scaling compensation compared to the surveying method or direct wand size compensation by a high precision 3D scanner. The presented validation methods can be less precise in some respects as compared to previous techniques, but they address an error type, which has not been and cannot be studied with the previous validation methods.     

Measuring ligament elasticity of the knee joint--elasticity measuring strip and its alternatives]

Those techniques for measuring ligament tension at the knee joint that are most commonly cited and easiest to carry out are discussed. These include four techniques based on the use of strain gauges. Apart from the Omega transducer and the buckle transducer, there is also the tendon force transducer, and the application of strain gauges to the bony ligament insertion sites. Other indirect measuring methods considered are the mercury strain transducer and the Hall effect transducer. The parameter measured with all of these methods is fluctuating current or voltage, which is then correlated with ligament tension. Three direct measurements are also discussed: the separation distances of marked fibres of the ligaments, replacement of fibres by threads, and a load cell/bone plug construction. The measured value is equated with the effective change in ligament length.     

Contribution of biomechanics to management of ligament and tendon injuries

The contribution of biomechanics to the advancement of management of ligament and tendon injuries has been significant. Thanks to Professor Y.C. Fung's writing and guidance, our field of research has done fundamental work on anatomy and biology of ligaments and tendons, developed methods to accurately determine mechanical properties, identified various experimental factors which could change the outcome measurements as well as examined biological factors that change tissue properties in-vivo. Professor Fung also gave us his quasi-linear viscoelastic theory for soft tissues so that the time and history dependent properties of ligaments and tendons could be properly described. We have further adopted Professor Fung's eight steps on methods of approach for biomechanical investigation to understand as well as enhance the treatment of ligament and tendon injuries during work or sports related activities. Examples on how to better treat the tears of the medial collateral ligament of the knee, as well as how to improve reconstruction procedures for the anterior cruciate ligament are presented in detail. Currently the use of functional tissue engineering for ligament and tendon healing is a topic of great interest. Here the use of biological scaffolds, such as porcine small intestinal submucosa, has shown promise. For the last 35 to 40 years, the field of biomechanics has made great strides in the treatment of ligament and tendon injuries, and many patients have benefited. The future is even brighter because of what has been done properly in the past. Exciting advances can be made in the field of tissue engineering through novel in-vitro culture and bioscaffold fabrication techniques. Recent technology can also allow the collection of in-vivo data so that ligament and tendon injuries can be better understood. Yet, solving new and more complex problems must still follow the stepwise methods of approach as taught by Professor Fung.     

Migration of radio-opaque markers injected into tendon grafts: a study using roentgen stereophotogrammetric analysis (RSA)

An increase in anterior laxity following reconstruction of the anterior cruciate ligament (ACL) can result from lengthening of the graft construct either at the sites of fixation and/or between the sites of fixation (i.e., graft substance). Roentgen stereophotogrammetric analysis (RSA), which requires that radio-opaque markers be attached to the graft, has been shown to be a useful technique in determining lengthening in these regions. Previous methods have been used for attaching radio-opaque markers to the graft, but they all have limitations particularly for single-loop grafts. Therefore, the objective of this study was to evaluate injecting markers directly into the substance of a tendon as a viable method for measuring lengthening of single-loop graft constructs by determining the maximum amount of migration after cyclic loading. Tantalum spheres of 0.8 mm diameter were used as tendon markers. Ten single-loop tendon grafts were passed through tibial tunnels drilled in calf tibias and fixed with a tibial fixation device. Two tendon markers were inserted in one tendon bundle of each graft and the grafts were cyclically loaded for 225,000 cycles from 20 N to 170 N. At specified intervals, simultaneous radiographs were obtained of the tendon markers. Marker migration was computed as the change in distance between the two tendon markers parallel to the axis of the tibial tunnel. Marker migration had a root mean square (RMS) value of less than 0.1 mm. Because the RMS value indicates the error introduced into measurements of lengthening and because this error is negligible, the method described for attaching markers to single-loop ACL grafts has the potential to be useful for determining lengthening of single-loop ACL graft constructs in in vivo studies in humans.     

Fiberoptic measurement of tendon forces is influenced by skin movement artifact

Fiberoptic cables have previously been used for tendon force measurements in vivo. To measure forces in the Achilles tendon, a cable is passed mediolaterally through the skin and tendon, transverse to the loading axis. As the tendon is loaded, its fibers compress the cable and modulate the intensity of transmitted light, which can be related to tendon force by an in situ calibration. The relative movement between skin and tendon at the cable entry and exit sites may cause error by bending the cable and thus altering transducer output. Cadaver simulations of walking were conducted to compare fiberoptic measurements of Achilles tendon forces to known loads applied to the tendon by actuators attached in series. Force measurement errors, which were high when the skin was intact (RMS errors 24-81% peak forces), decreased considerably after skin removal (RMS errors 10-33% peak forces). The fiberoptic transducer is a useful tool for measurement of tendon forces in situ under natural loading conditions when skin can be removed, but caution should be exercised during in vivo use of this technique or under circumstances where skin is in contact with the fiberoptic cable at the insertion and exit sites.     

A method to characterize in vivo tendon force-strain relationship by combining ultrasonography, motion capture and loading rates

The ultrasonography contributes to investigate in vivo tendon force-strain relationship during isometric contraction. In previous studies, different methods are available to estimate the tendon strain, using different loading rates and models to fit the tendon force-strain relationship. This study was aimed to propose a standard method to characterize the in vivo tendon force-strain relationship. We investigated the influence on the force-strain relationship for medialis gastrocnemius (MG) of (1) one method which takes into account probe and joint movements to estimate the instantaneous tendon length, (2) models used to fit the force-strain relationship for uniaxial test (polynomial vs. Ogden), and (3) the loading rate on tendon strain. Subjects performed ramp-up contraction during isometric contractions at two different target speeds: 1.5s and minimal time with ultrasound probe fixed over the muscle-tendon junction of the MG muscle. The used method requires three markers on ultrasound probe and a marker on calcaneum to take into account all movements, and was compared to the strain estimated using ultrasound images only. The method using ultrasound image only overestimated the tendon strain from 40% of maximal force. The polynomial model showed similar fitting results than the Ogden model (R2=0.98). A loading rate effect was found on tendon strain, showing a higher strain when loading rate decreases. The characterization of tendon force-strain relationship needs to be standardized by taking into account all movements to estimate tendon strain and controlling the loading rate. The polynomial model appears to be appropriate to represent the tendon force-strain relationship.     

Form and function of the oblique cord (chorda obliqua) in anthropoid primates

The oblique cord of the forearm in humans is a ligament connecting the anterolateral aspect of the ulna proximally to the posteromedial aspect of the radius distally, inserting just below the radial tuberosity. Its functional significance is uncertain, but it has been proposed that the ligament limits supination of the forearm or aids in resisting buckling failure in the curved radius. These functional explanations are unsatisfactory for bipedal humans who do not use their forelimbs in weight support. Furthermore, there are no evolutionary explanations for its presence in humans. The purpose of this study was to investigate the distribution and form of the oblique cord in non-human primates and to explore its possible functional significance and evolutionary origin in humans. Soft tissue dissections of anthropoid primates revealed the presence of an oblique cord in New and Old World monkeys and Asian apes. It was absent in all atelines. Passive manipulation of the two forearm bones in all specimens revealed that the ligament becomes most taut in pronation contrary to the proposed human condition. Isolated radii show that the oblique cord's radial insertion lacks a clear relationship with bone curvature, thus providing little support for the hypothesis of preventing buckling failure. The oblique cord's involvement in reducing bending strain in the curved radius is also unlikely. It is suggested here that the ligament may have a role in maintaining elbow stability in quadrupedal primates. Therefore, the function of the oblique cord in anthropoid primates suggests that its presence in modern humans may be a morphological and functional remnant from a quadrupedal ancestry.     

Evaluation of different analytical methods for subject-specific scaling of musculotendon parameters

Musculoskeletal models are often used to estimate internal muscle forces and the effects of those forces on the development of human movement. The Hill-type muscle model is an important component of many of these models, yet it requires specific knowledge of several muscle and tendon properties. These include the optimal muscle fibre length, the length at which the muscle can generate maximum force, and the tendon slack length, the length at which the tendon starts to generate a resistive force to stretch. Both of these parameters greatly influence the force-generating behaviour of a musculotendon unit and vary with the size of the person. However, these are difficult to measure directly and are often estimated using the results of cadaver studies, which do not account for differences in subject size. This paper examined several different techniques that can be used to scale the optimal muscle fibre length and tendon slack length of a musculotendon unit according to subject size. The techniques were divided into three categories corresponding to linear scaling, scaling by maintaining a constant tendon slack length throughout the range of joint motion, and scaling by maintaining muscle operating range throughout the range of joint motion. We suggest that a good rationale for scaling muscle properties should be to maintain the same force-generating characteristics of a musculotendon unit for all subjects, which is best achieved by scaling that preserves the muscle operating range when the muscle is maximally activated.     

Tendon strain measurements with dynamic ultrasound images: evaluation of digital image correlation

Strain is an essential metric in tissue mechanics. Strains and strain distributions during functional loads can help identify damaged and pathologic regions as well as quantify functional compromise. Noninvasive strain measurement in vivo is difficult to perform. The goal of this in vitro study is to determine the efficacy of digital image correlation (DIC) methods to measure strain in B-mode ultrasound images. The Achilles tendons of eight male Wistar rats were removed and mechanically cycled between 0 and 1% strain. Three cine video images were captured for each specimen: (1) optical video for manual tracking of optical markers; (2) optical video for DIC tracking of optical surface markers; and (3) ultrasound video for DIC tracking of image texture within the tissue. All three imaging modalities were similarly able to measure tendon strain during cyclic testing. Manual/ImageJ-based strain values linearly correlated with DIC (optical marker)-based strain values for all eight tendons with a slope of 0.970. DIC (optical marker)-based strain values linearly correlated with DIC (ultrasound texture)-based strain values for all eight tendons with a slope of 1.003. Strain measurement using DIC was as accurate as manual image tracking methods, and DIC tracking was equally accurate when tracking ultrasound texture as when tracking optical markers. This study supports the use of DIC to calculate strains directly from the texture present in standard B-mode ultrasound images and supports the use of DIC for in vivo strain measurement using ultrasound images without additional markers, either artificially placed (for optical tracking) or anatomically in view (i.e., bony landmarks and/or muscle-tendon junctions).     

Bioactive nanofibers for fibroblastic differentiation of mesenchymal precursor cells for ligament/tendon tissue engineering applications

Mesenchymal stem cells and precursor cells are ideal candidates for tendon and ligament tissue engineering; however, for the stem cell-based approach to succeed, these cells would be required to proliferate and differentiate into tendon/ligament fibroblasts on the tissue engineering scaffold. Among the various fiber-based scaffolds that have been used in tendon/ligament tissue engineering, hybrid fibrous scaffolds comprising both microfibers and nanofibers have been recently shown to be particularly promising. With the nanofibrous coating presenting a biomimetic surface, the scaffolds can also potentially mimic the natural extracellular matrix in function by acting as a depot for sustained release of growth factors. In this study, we demonstrate that basic fibroblast growth factor (bFGF) could be successfully incorporated, randomly dispersed within blend-electrospun nanofibers and released in a bioactive form over 1 week. The released bioactive bFGF activated tyrosine phosphorylation signaling within seeded BMSCs. The bFGF-releasing nanofibrous scaffolds facilitated BMSC proliferation, upregulated gene expression of tendon/ligament-specific ECM proteins, increased production and deposition of collagen and tenascin-C, reduced multipotency of the BMSCs and induced tendon/ligament-like fibroblastic differentiation, indicating their potential in tendon/ligament tissue engineering applications.     

Scaling in tensile "skeletons": scale dependent length of the Achilles tendon in mammals

The Achilles tendon of a diverse group of mammals ranging from the mouse (12g) to the rhinoceros (1300kg) scales so that the tendon length varies as tendon diameter.^0-931±0.069 ( r =0.983). Tendon length scales as (body mass)^0-342±0.028, and tendon diameter scales as (body mass)^0-361±0.029. If tendon stress and strain are scale independent, the capacity of the tendon to store elastic strain energy remains proportion to body mass. If tendon stress and strain increase with body mass, energy storage may scale somewhat higher. The scaling of the Achilles tendon is consistent with its role in storing strain energy and different from that of a variety of other tensile skeletal elements which exhibit scale independent length dimensions.     

Predicting the external formation of callus tissues in oblique bone fractures: idealised and clinical case studies

It is proposed that the external asymmetric formation of callus tissues that forms naturally about an oblique bone fracture can be predicted computationally. We present an analysis of callus formation for two cases of bone fracture healing: idealised and subject-specific oblique bone fractures. Plane strain finite element (FE) models of the oblique fractures were generated to calculate the compressive strain field experienced by the immature callus tissues due to interfragmentary motion. The external formations of the calluses were phenomenologically simulated using an optimisation style algorithm that iteratively removes tissue that experiences low strains from a large domain. The resultant simulated spatial formation of the healing tissues for the two bone fracture cases showed that the calluses tended to form at an angle equivalent to the angle of the oblique fracture line. The computational results qualitatively correlated with the callus formations found in vivo. Consequently, the proposed methods show potential as a means of predicting callus formation in pre-clinical testing.     

Modeling short-range stiffness of feline lower hindlimb muscles

The short-range stiffness (SRS) of skeletal muscles is a critical property for understanding muscle contributions to limb stability, since it represents a muscle's capacity to resist external perturbations before reflexes or voluntary actions can intervene. A number of studies have demonstrated that a simple model, consisting of a force-dependent active stiffness connected in series with a constant passive stiffness, is sufficient to characterize the SRS of individual muscles over the entire range of obtainable forces. The purpose of this study was to determine if such a model could be used to characterize the SRS-force relationship in a number of architecturally distinct muscles. Specifically, we hypothesized that the active and passive stiffness components for a specific muscle can be estimated from anatomical measurements, assuming uniform active and passive stiffness properties across all muscles. This hypothesis was evaluated in six feline lower hindlimb muscle types with different motor unit compositions and architectures. The SRS-force relationships for each muscle type were predicted based on anatomical measurements and compared to experimental data. The model predictions were accurate to within 30%, when uniform scaling properties were assumed across all muscles. Errors were the greatest for the extensor digitorum longus (EDL). When this muscle was removed from the analysis, prediction errors dropped to less than 8%. Subsequent analyses suggested that these errors might have resulted from differences in the tendon elastic modulus, as compared to the other muscles tested.