In vitro biomechanical properties of sole tissues: Comparison between healthy and ulcerated bovine claws.

Sole ulcers are reportedly one of the most prevalent diseases associated with lameness in dairy cattle, significantly affecting animal welfare and farm profitability. The degree to which sole soft tissues, healthy or ulcerated, are able to maintain their structure and function when subjected to compressive forces remains unknown. Therefore, the aims of the present study were to assess sole tissue biomechanics in healthy and ulcerated claws and to describe correlated histology. Cylindrical samples were harvested from zones 4 and 6, as described by the international foot map, from hind lateral healthy (n = 12) and ulcerated bovine claws (n = 8; animals n = 12). Tissue biomechanics and morphology were evaluated via compressive tests and hematoxylin-eosin-phloxine-saffron staining, respectively. A 2-sample t-test was used to compare zones' mechanical properties between healthy and ulcerated tissues, and the Cochran-Mantel-Haenszel test was used to measure the effect of claw zone on histology. The fibril modulus (Ef) and permeability (k) respectively increased and decreased in ulcerated claws (Ef = 0.201 ± 0.104 MPa; k = 0.128 ± 0.069 mm2/MPa·s) compared with healthy claws (Ef = 0.105 ± 0.050 MPa; k = 0.452 ± 0.365 mm2/MPa·s) only for zone 6. Histology scores equal to or greater than 3 were associated with macroscopic presence of ulceration. A higher proportion of adipose tissue (30% or more) was associated with zone 6 compared with zone 4, but no difference was seen between healthy and ulcerated claws. Ulcerated claws had a higher prevalence of exostoses compared with healthy ones (33% vs. 8%). Sole soft tissues showed, as hypothesized, a viscoelastic behavior using unconfined compression testing, which, however, may not reflect in vivo loading conditions. Clinical and histological signs of sole ulceration were not associated with decreased strength of the supportive apparatus of the distal phalanx in zone 4 in this study.

In vivo dynamic compression has less detrimental effect than static compression on newly formed bone of a rat caudal vertebra.

Fusionless devices are currently designed to treat spinal deformities such as scoliosis by the application of a controlled mechanical loading. Growth modulation by dynamic compression was shown to preserve soft tissues. The objective of this in vivo study was to characterize the effect of static vs. dynamic loading on the bone formed during growth modulation. Controlled compression was applied during 15 days on the 7(th) caudal vertebra (Cd7) of rats during growth spurt. The load was sustained in the "static" group and sinusoidally oscillating in the "dynamic" group. The effect of surgery and of the device was investigated using control and sham (operated on but no load applied) groups. A high resolution CT-scan of Cd7 was acquired at days 2, 8 and 15 of compression. Growth rates, histomorphometric parameters and mineral density of the newly formed bone were quantified and compared. Static and dynamic loadings significantly reduced the growth rate by 20% compared to the sham group. Dynamic loading preserved newly formed bone histomorphometry and mineral density whereas static loading induced thicker (+31%) and more mineralized (+12%) trabeculae. A significant sham effect was observed. Growth modulation by dynamic compression constitutes a promising way to develop new treatment for skeletal deformities.

An automated cell viability quantification method for low-resolution confocal images of closely packed cells based on a modified gradient flow tracking algorithm.

Fluorescent-based live/dead labelling combined with fluorescent microscopy is one of the widely used and reliable methods for assessment of cell viability. This method is, however, not quantitative. Many image-processing methods have been proposed for cell quantification in an image. Among all these methods, several of them are capable of quantifying the number of cells in high-resolution images with closely packed cells. However, no method has addressed the quantification of the number of cells in low-resolution images containing closely packed cells with variable sizes. This paper presents a novel method for automatic quantification of live/dead cells in 2D fluorescent low-resolution images containing closely packed cells with variable sizes using a mean shift-based gradient flow tracking. Accuracy and performance of the method was tested on growth plate confocal images. Experimental results show that our algorithm has a better performance in comparison to other methods used in similar detection conditions.

Biomechanical analysis of fracture risk associated with tibia deformity in children with osteogenesis imperfecta: a finite element analysis.

OBJECTIVES: Osteogenesis imperfecta (OI) frequently leads to long-bone bowing requiring a surgical intervention in severe cases to avoid subsequent fractures. However, there are no objective criteria to decide when to perform such intervention. The objective is to develop a finite element model to predict the risk of tibial fracture associated with tibia deformity in patients with OI. METHODS: A comprehensive FE model of the tibia was adapted to match bi-planar radiographs of a 7 year-old girl with OI. Ten additional models with different deformed geometries (from 2° to 24°) were created and the elasto-plastic mechanical properties were adapted to reflect OI conditions. Loads were obtained from mechanography of two-legged hopping. Two additional impact cases (lateral and torsion) were also simulated. Principal strain levels were used to define a risk criterion. RESULTS: Fracture risks for the two-legged hopping load case remained low and constant until tibia bowing reached 15° and 16° in sagittal and coronal planes respectively. Fracture risks for lateral and torsion impact were equivalent whatever the level of tibial bowing. CONCLUSIONS: The finite element model of OI tibia provides an objective means of assessing the necessity of surgical intervention for a given level of tibia bowing in OI-affected children.

Biomechanical modeling of the lateral decubitus posture during corrective scoliosis surgery.

BACKGROUND: Patient prone positioning in scoliosis surgeries modifies the spinal curves prior to instrumentation. However, the biomechanical effects of the lateral decubitus posture, used in anterior approaches and minimally invasive techniques, have not yet been investigated. The objectives were to develop and validate a finite element model simulating the spinal changes resulting from this positioning. METHODS: The 3D pre-op reconstructed geometries of six adolescent patients with idiopathic scoliosis were used to develop personalized finite element models of the spine, which integrated a three-step method simulating the lateral posture. Clinical indices were measured on pre- and intra-operative radiographs to validate the finite element model. FINDINGS: The major Cobb angle and apical vertebral translation were reduced by 44% and 37% respectively between the pre- and intra-op postures. Using appropriately oriented gravity forces and boundary conditions, the finite element model simulations represented adequately these changes, with average differences of 4 degrees for the major Cobb angle and 4mm for the apical vertebral translation with the radiographic values. INTERPRETATION: Lateral decubitus positioning significantly reduces the spinal deformities prior to instrumentation, as demonstrated by the finite element model. This study is a first step in the development of a modeling tool for the optimal adjustments of intra-operative positioning, which remains to be further investigated with complementary clinical studies.

Mechanical loading effects on isthmic spondylolytic lumbar segment: finite element modelling using a personalised geometry.

Biomechanics of the isthmic spondylolysis was investigated by using a nonlinear 3D-finite element model (FEM). A personalised in vivo pediatric geometry of L5-S1 low-grade spondylolisthesis patient was used to develop a L5-pelvis motion segment model that took into consideration vertebrae, disc and ligaments. The stress distribution in the affected motion segment under axial force only, and for a combination of flexion and extension was evaluated. Predicted results showed that, under all loading conditions, stresses were much higher on the pedicle and in the dorsal wall of the pars interarticularis due to the abnormal geometry which is consistent with clinical observations.

Finite element modeling of vertebral body stapling applied for the correction of idiopathic scoliosis: preliminary results.

Endoscopic vertebral body stapling is an innovative technique intended to treat adolescent idiopathic scoliosis, but the optimal instrumentation design is not yet established. The objective was to simulate the immediate correction obtained from two stapling configurations. A parametric finite element model of a typical right thoracic scoliotic spine (Cobb 21 degrees ) was developed using geometrical and mechanical data from the literature. Staple insertion and closing were modeled. The intra-operative lateral decubitus and standing positions were taken into account. Two implant configurations, varying the number of staples per vertebra, were simulated. The major correction (9 degrees ) came by simulating the intra-operative posture. The immediate Cobb angle correction due to the staples alone was less then 1 degrees for both configurations. However, the staples helped maintain the correction obtained by the intra-operative posture when the post-operative standing position was simulated. Next steps are to validate the model using surgical cases, implement growth modulation modeling, improve lateral decubitus modeling, and analyze different vertebral stapling strategies for different scoliotic curves.

Non-uniform strain distribution within rat cartilaginous growth plate under uniaxial compression.

Growth plates are highly inhomogeneous in morphology and composition. Mechanical loading can modulate longitudinal bone growth, though the mechanisms underlying this mechanobiology are poorly understood. The proximal tibial growth plates of six rats were tested in vitro under uniaxial compression to 5% strain, and confocal microscopy was used to track and capture images of fluorescently labeled cell nuclei with increasing applied strains. The local strain patterns through the growth plate thickness were quantified using texture correlation analysis. The technique of texture correlation analysis was first validated by comparing theoretical simulated strain maps generated from numerically distorted images. The texture correlation algorithm was sensitive to the grid size superimposed on the original image, but remained insensitive to parameters related to the size of the final image mask, which was searched by the correlation algorithm for each grid point of the original image. Within the growth plate, experimental strain distributions were non-uniform in all six specimens. Growth plates were mostly under compression strains. The strain distributions differed among the histomorphological zones of the growth plate, which was most obvious in specimens with regular growth plate shape: higher compressive strains (4-8 times higher than the applied 5% strain) were located mainly in regions overlapping the reserve and hypertrophic zones with lower compressive strains in the proliferative zone. This study documents the non-uniform mechanical behavior of growth plate across its three histological zones when exposed to compression. Further investigation is required to establish the significance of non-uniform strain fields during growth in vivo.

Static compressive loading reduces the mRNA expression of type II and X collagen in rat growth-plate chondrocytes during postnatal growth.

The mechanisms by which chondrocytes modulate longitudinal bone growth are not well understood. This in vitro study investigated the effects of loading on the mRNA expression pattern of key molecular components of the growth-plate related to the extracellular matrix (type II and type X collagen) and the PTH-PTHrP feedback loop. Short-term static compressive loading was applied to rat proximal tibial growth-plate explants. Four age groups at specific developmental stages were investigated. The spatial variation in the mRNA expression was compared among loaded explants, their contralateral sham controls, and uncultured growth plates from normal animals. Basic cell metabolism (18S rRNA) was unaffected by load. Results indicated a narrower spatial distribution of mRNA expression of type II collagen throughout the growth plate; similarly, a narrowed distribution of expression of type X collagen was noted in the lower hypertrophic zone of the growth-plate. This suggests that mechanical compression influences chondrocytes of the hypertrophic zone to alter their expression of specific genes encoding proteins of the extracellular matrix, while PTH-PTHrP receptor mRNA, a regulatory protein, remained unaffected by loading. The effects of compression were similar at the different stages of growth, suggesting that additional factors may be involved in the clinical progression of skeletal deformities observed during growth spurts. Although this study was done in vitro and limited to static loading, it furthers our understanding of growth-plate mechanobiology as a first step toward providing a scientific rationale for treating progressive musculoskeletal deformities.

Biomechanical modelling of growth modulation following rib shortening or lengthening in adolescent idiopathic scoliosis.

A biomechanical model was developed to evaluate the long-term correction resulting from rib shortening or lengthening in adolescent idiopathic scoliosis (AIS). A finite element model of the trunk, personalised to the geometry of a scoliotic patient, was used to simulate rib surgery. Stress relaxation of ligaments following surgery was integrated into the model, as well as longitudinal growth of vertebral bodies and ribs and its modulation due to mechanical stresses. Simulations were performed in an iterative fashion over 24 months. A concave side rib shortening, inducing load patterns on the vertebral end-plates that could act against the scoliosis progression, was tested. A fractional factorial experimental design of 16 runs documented the effects of six modelling parameters. Wedging of the apical vertebra in the frontal plane decreased from 5.2 degrees initially to a mean value of 3.8 degrees after 24 months. The wedging decrease in the thoracic apical region was reflected by changes in the spine curvature, with a Cobb angle decrease from 46 degrees to 44 degrees immediately after the surgery and to a mean of 41 degrees after 24 months. However, both rib hump and vertebral axial rotation increased, on average, by 4 degrees at the curve apex. The most significant parameters were the growth sensitivity to stress in ribs and vertebrae and the rate of stress relaxation of intercostal ligaments. The results confirmed the potential of long-term correction of spinal curvature resulting from the rib shortening on the concavity. This modelling approach could be used for further design of less invasive surgery, taking into account residual growth, for scoliosis correction.

Biomechanical simulations of the spine deformation process in adolescent idiopathic scoliosis from different pathogenesis hypotheses.

It is generally recognized that progressive adolescent idiopathic scoliosis (AIS) evolves within a self-sustaining biomechanical process involving asymmetrical growth modulation of vertebrae due to altered spinal load distribution. A biomechanical finite element model of normal thoracic and lumbar spine integrating vertebral growth was used to simulate the progression of spinal deformities over 24 months. Five pathogenesis hypotheses of AIS were represented, using an initial geometrical eccentricity (gravity line imbalance of 3 mm or 2 degrees rotation) at the thoracic apex to trigger the self-sustaining deformation process. For each simulation, regional (thoracic Cobb angle, kyphosis) and local scoliotic descriptors (axial rotation and wedging of the thoracic apical vertebra) were evaluated at each growth cycle. The simulated AIS pathogeneses resulted in the development of different scoliotic deformities. Imbalance of 3 mm in the frontal plane, combined or not with the sagittal plane, resulted in the closest representation of typical scoliotic deformities, with the thoracic Cobb angle progressing up to 39 degrees (26 degrees when a sagittal offset was added). The apical vertebral rotation increased by 7 degrees towards the convexity of the curve, while the apical wedging increased to 8.5 degrees (7.3 degrees with the sagittal eccentricity) and this deformity evolved towards the vertebral frontal plane. A sole eccentricity in the sagittal plane generated a non-significant frontal plane deformity. Simulations involving an initial rotational shift (2 degrees ) in the transverse plane globally produced relatively small and non-typical scoliotic deformations. Overall, the thoracic segment predominantly was sensitive to imbalances in the frontal plane, although unidirectional geometrical eccentricities in different planes produced three-dimensional deformities at the regional and vertebral levels, and their deformities did not cumulate when combined. These results support the hypothesis of a prime lesion involving the precarious balance in the frontal plane, which could concomitantly be associated with a hypokyphotic component. They also suggest that coupling mechanisms are involved in the deformation process.

Simulation of progressive deformities in adolescent idiopathic scoliosis using a biomechanical model integrating vertebral growth modulation.

While the etiology and pathogenesis of adolescent idiopathic scoliosis are still not well understood, it is generally recognized that it progresses within a biomechanical process involving asymmetrical loading of the spine and vertebral growth modulation. This study intends to develop a finite element model incorporating vertebral growth and growth modulation in order to represent the progression of scoliotic deformities. The biomechanical model was based on experimental and clinical observations, and was formulated with variables integrating a biomechanical stimulus of growth modulation along directions perpendicular (x) and parallel (y, z) to the growth plates, a sensitivity factor beta to that stimulus and time. It was integrated into a finite element model of the thoracic and lumbar spine, which was personalized to the geometry of a female subject without spinal deformity. An imbalance of 2 mm in the right direction at the 8th thoracic vertebra was imposed and two simulations were performed: one with only growth modulation perpendicular to growth plates (Sim1), and the other one with additional components in the transverse plane (Sim2). Semi-quantitative characterization of the scoliotic deformities at each growth cycle was made using regional scoliotic descriptors (thoracic Cobb angle and kyphosis) and local scoliotic descriptors (wedging angle and axial rotation of the thoracic apical vertebra). In all simulations, spinal profiles corresponded to clinically observable configurations. The Cobb angle increased non-linearly from 0.3 degree to 34 degrees (Sim1) and 20 degrees (Sim2) from the first to last growth cycle, adequately reproducing the amplifying thoracic scoliotic curve. The sagittal thoracic profile (kyphosis) remained quite constant. Similarly to clinical and experimental observations, vertebral wedging angle of the thoracic apex progressed from 2.6 degrees to 10.7 degrees (Sim1) and 7.8 degrees (Sim2) with curve progression. Concomitantly, vertebral rotation of the thoracic apex increased of 10 degrees (Sim1) and 6 degrees (Sim2) clockwise, adequately reproducing the evolution of axial rotation reported in several studies. Similar trends but of lesser magnitude (Sim2) suggests that growth modulation parallel to growth plates tend to counteract the growth modulation effects in longitudinal direction. Overall, the developed model adequately represents the self-sustaining progression of vertebral and spinal scoliotic deformities. This study demonstrates the feasibility of the modeling approach, and compared to other biomechanical studies of scoliosis it achieves a more complete representation of the scoliotic spine.

Correlation study between indices describing the scoliotic spine.

This study intended to investigate the correlations between different geometrical indices in order to assess their usefulness in the characterization of scoliotic deformities. Analytical evaluation of indices was obtained from 3D reconstruction of vertebrae. Scoliotic indices included the Cobb angle, the Cobb angle in the plane of maximum curvature, the angular orientation of the plane of maximum curvature, the kyphosis and the maximal axial rotation. This analysis was applied to the thoracic curve of 100 scoliotic adolescents. The correlations between these five indices were separately investigated for the RT and RTLL curve types. A statistical correlation was found between the angular orientation of the plane of maximum curvature and the kyphosis. The results indicate a relative independence between most indices. Hence, an evaluation of several complementary indices is required to provide a more complete description of 3D scoliotic deformities.

Biomechanical modelling of spinal growth modulation for the study of adolescent scoliotic deformities: a feasibility study.

A model of growth modulation was formulated with variables integrating a biomechanical stimulus of growth modulation, a sensitivity factor to the stimulus and time. It was integrated into a finite element model of the thoracic and lumbar spine using an iterative procedure. A simulation on the personalized geometry of a mild scoliotic patient allowed qualitative investigation of scoliotic deformities over 12 cycles (months) in response to a load variation due to an eccentricity of the patient's gravity line in the frontal plane. Resulting frontal, sagittal and transverse spinal views correspond to clinically observable scoliotic configurations. The simulation adequately reproduces a progressing thoracic scoliotic curve while the slight increasing kyphosis represents a possible condition although a thoracic hypokyphosis is frequently reported in the literature. At the thoracic apex, increased wedging as well as axial rotation evolving towards curve convexity are in agreement with clinical and experimental observations reported with curve progression. This study demonstrates the feasibility of the approach and, compared to other biomechanical models, it achieves a more complete representation of the scoliotic spine by incorporating vertebral growth modulation.

Evolution of 3D deformities in adolescents with progressive idiopathic scoliosis.

The objective of this study was to conduct an intrasubject longitudinal study quantifying the evolution of two- and three-dimensional geometrical scoliotic descriptors. The evolution of regional and local scoliotic descriptors was analyzed between two scoliotic visits on a cohort of 28 adolescents with progressive idiopathic scoliosis. Mean age at the first visit was 12.7 +/- 1.7 years old and averaged time interval between two assessments reached 22.8 +/- 10.8 months. Scoliotic descriptors were obtained from three-dimensionally reconstructed spines. The initial thoracic Cobb angle was on average 35.3 degrees +/- 8.4 degrees (range, 14 degrees-54 degrees). The evolution of spinal curvatures and vertebral deformities was assessed statistically in terms of descriptor absolute variations, and of descriptor variations normalized with respect to time and to the increase in Cobb angle. At the thoracic level, vertebral wedging increased with curve severity in a relatively consistent pattern for most scoliotic patients and axial rotation mainly increased towards curve convexity with scoliosis severity. No consistent evolution was associated with the angular orientation of the maximum wedging. Thoracic kyphosis changes (increase and decrease) were observed in important proportions. Results of this study challenge the existence of a typical scoliotic evolution pattern and suggest that the scoliotic evolution is quite variable and patient-specific.

The effects of mechanical loading on the mRNA expression of growth-plate cells.

Bone growth is a complex process involving proliferation, maturation and hypertrophy of chondrocytes in the growth plates. Mechanical forces applied to growing bones alter their longitudinal growth. However, the mechanisms by which chondrocytes modulate longitudinal bone growth are not well understood. This in vitro study investigated the effects of mechanical loading on the mRNA expression pattern of key molecular components of the growth-plate. Short-term static loading was applied to rat proximal tibial growth-plate explants. Various age groups at specific developmental stages were investigated. In situ hybridization was used to assess the mRNA expression of the cells in different zones of the growth-plate. Four key components were investigated: 18s (basic cell metabolism), type II collagen (major extracellular matrix component), type X collagen (matrix component in hypertrophic zone) and PTH-PTHrP receptors (pre-hypertrophic chondrocytes). The spatial variation in the mRNA expression between loaded explants and their contralateral controls was compared to establish: -the sensitivity of the different growth-plate zones to mechanical loading; -the sensitivity of the different developmental stages to loading. Preliminary results indicated that static loading on the growth plate of 80 d.o. rats affects type II and X collagen gene expressions while PTH-PTHrP remains insensitive to static loading. Improved understanding of growth-plate mechanics and the underlying biology is required to provide a scientific basis for the treatment of progressive deformities.

Progression of vertebral and spinal three-dimensional deformities in adolescent idiopathic scoliosis: a longitudinal study.

STUDY DESIGN: The evolution of scoliotic descriptors was analyzed from three-dimensionally reconstructed spines and assessed statistically in a group of adolescents with progressive idiopathic scoliosis. OBJECTIVES: To conduct an intrasubject longitudinal study quantifying evolution of two- and three-dimensional geometrical descriptors characterizing the scoliotic spine and vertebral deformities. SUMMARY OF BACKGROUND DATA: The data available on geometric descriptors usually are based on cross-sectional studies comparing scoliotic configurations of different individuals. The literature reports very few longitudinal studies that evaluated different phases of scoliotic progression in the same patients. METHODS: The evolution of regional and local descriptors between two scoliotic visits was analyzed in 28 adolescents with scoliosis. Several statistical analyses were performed to determine how spinal curvatures and vertebral deformities change during scoliosis progression. RESULTS: At the thoracic level, vertebral wedging increases with curve severity in a relatively consistent pattern for most patients with scoliosis. Axial rotation mainly increases toward curve convexity with scoliosis severity, worsening the progression of vertebral body deformities. No consistent evolution is associated with the angular orientation of the maximum wedging. Thoracic kyphosis varies considerably among subjects. Both increasing and decreasing kyphosis are observed in nonnegligible proportions. A decrease in kyphosis is associated with a shift in the plane of maximum deformity toward the frontal plane, which worsens the three-dimensional shape of the spine. CONCLUSIONS: The results of this study challenge the existence of a typical scoliotic evolution pattern and suggest that scoliotic evolution is quite variable and patient specific.

[Correlation study between spinal curvatures and vertebral and disk deformities in idiopathic scoliosis]. / Etude des relations entre les courbures rachidiennes et les déformations vertébrales et discales dans la scoliose idiopathique.

Idiopathic scoliosis involves complex tridimensional (3D) deformations of the spine associated with intrinsic alterations (wedging) of vertebral bodies (VB) and intervertebral disks (ID). This study intends to evaluate analytically in vivo 2D and 3D scoliotic descriptors, based on clinical data from 40 thoracic curves of scoliotic adolescents, and to establish relationships between the regional curve deformations and the local VB and ID deformities. A multiplanar radiographic technique provided 3D positioning of vertebral landmarks. Cobb angle in the postero-anterior (PA) view, in the plane of maximum deformity (CobbP.Max) and the angular orientation of the plane of maximum deformity were used as regional descriptors. Vertebral body endplates were modeled as 3D oriented ellipses. Axial rotation, global PA and local frontal wedgings (inclinations of projected ellipses in the global and vertebral frontal planes), 3D maximum wedging (real inclination of adjacent ellipses) as well as the angular orientation of 3D wedging were calculated to characterize local deformations at the thoracic apex. Mean values for CobbPA, CobbP.Max and the angular orientation of the maximum deformity (with respect to the sagittal plane) reached 44 degrees, 48 degrees and 67 degrees respectively. On average, vertebral axial rotation, global PA, local frontal and 3D wedging angles were respectively 15 degrees, 8.3 degrees, 8.2 degrees and 9.7 degrees. Analyses indicated statistical correlation between: a) Cobb angles and vertebral wedging; b) the orientations of the maximum deformity and of 3D vertebral wedging; c) the axial rotation and CobbPA; d) the axial rotation and the angular orientation of 3D vertebral wedging. At the thoracic level, statistical analyses indicated that vertebral wedging and axial rotation increase with curve progression. Scoliosis severity, as measured by Cobb angles, evolves simultaneously to a coronalization of the plane of maximum deformity, revealing an hypokyphotic phenomenon, and to a real vertebral wedging shifting towards the frontal plane of the vertebra. These 3D in vivo analyses allowed interpretation of spatial relationships between regional and local scoliotic deformities. Compared to 2D in vivo or 3D in vitro analyses alone, this 3D in vivo study provides a more complete assessment of spinal curve progression to fully interpret the real 3D curvature and intrinsic deformations as well as their evolution processes.