Injectable bioactive poly(propylene fumarate) and polycaprolactone based click chemistry bone cement for spinal fusion in rabbits.

Degenerative spinal pathology is a widespread medical issue, and spine fusion surgeries are frequently performed. In this study, we fabricated an injectable bioactive click chemistry polymer cement for use in spinal fusion and bone regrowth. Taking advantages of the bioorthogonal click reaction, this cement can be crosslinked by itself eliminating the addition of a toxic initiator or catalyst, nor any external energy sources like UV light or heat. Furthermore, nano-hydroxyapatite (nHA) and microspheres carrying recombinant human bone morphogenetic protein-2 (rhBMP-2) and recombinant human vascular endothelial growth factor (rhVEGF) were used to make the cement bioactive for vascular induction and osteointegration. After implantation into a rabbit posterolateral spinal fusion (PLF) model, the cement showed excellent induction of new bone formation and bridging bone, achieving results comparable to autograft control. This is largely due to the osteogenic properties of nano-hydroxyapatite (nHA) and the released rhBMP-2 and rhVEGF growth factors. Since the availability of autograft sources is limited in clinical settings, this injectable bioactive click chemistry cement may be a promising alternative for spine fusion applications in addressing various spinal conditions.

Bioactive Moldable Click Chemistry Polymer Cement with Nano-Hydroxyapatite and Growth Factor-Enhanced Posterolateral Spinal Fusion in a Rabbit Model.

Spinal injuries or diseases necessitate effective fusion solutions, and common clinical approaches involve autografts, allografts, and various bone matrix products, each with limitations. To address these challenges, we developed an innovative moldable click chemistry polymer cement that can be shaped by hand and self-cross-linked in situ for spinal fusion. This self-cross-linking cement, enabled by the bioorthogonal click reaction, excludes the need for toxic initiators or external energy sources. The bioactivity of the cement was promoted by incorporating nanohydroxyapatite and microspheres loaded with recombinant human bone morphogenetic protein-2 and vascular endothelial growth factor, fostering vascular induction and osteointegration. The release kinetics of growth factors, mechanical properties of the cement, and the ability of the scaffold to support in vitro cell proliferation and differentiation were evaluated. In a rabbit posterolateral spinal fusion model, the moldable cement exhibited remarkable induction of bone regeneration and effective bridging of spine vertebral bodies. This bioactive moldable click polymer cement therefore presents a promising biomaterial for spinal fusion augmentation, offering advantages in safety, ease of application, and enhanced bone regrowth.

Extrusion 3D-printing and characterization of poly(caprolactone fumarate) for bone regeneration applications.

Polycaprolactone fumarate (PCLF) is a cross-linkable PCL derivative extensively considered for tissue engineering applications. Although injection molding has been widely used to develop PCLF scaffolds, platforms developed using such technique lack precise control on architecture, design, and porosity required to ensure adequate cellular and tissue responses. In particular, the scaffolds should provide a suitable surface for cell attachment and proliferation, and facilitate cell-cell communication and nutrient flow. 3D printing technologies have led to new architype for biomaterial development with micro-architecture mimicking native tissue. Here, we developed a method for 3D printing of PCLF structures using the extrusion printing technique. The crosslinking property of PCLF enabled the unique post-processing of 3D printed scaffolds resulting in highly porous and flexible PCLF scaffolds with compressive properties imitating natural features of cancellous bone. Generated scaffolds supported excellent attachment and proliferation of mesenchymal stem cells (MSC). The high porosity of PCLF scaffolds facilitated vascularized membrane formation demonstrable with the stringency of the ex ovo chicken chorioallantoic membrane (CAM) implantation. Furthermore, upon implantation to rat calvarium defects, PCLF scaffolds enabled an exceptional new bone formation with a bone mineral density of newly formed bone mirroring native bone tissue. These studies suggest that the 3D-printed highly porous PCLF scaffolds may serve as a suitable biomaterial platform to significantly expand the utility of the PCLF biomaterial for bone tissue engineering applications.

Bioorthogonal "Click Chemistry" Bone Cement with Bioinspired Natural Mimicking Microstructures for Bone Repair.

Current bone cement systems often demand free radical or metal-related initiators and/or catalysts for the crosslinking process, which may cause serious toxicity to the human body. In addition, the resultant dense scaffolds may have a prolonged degradation time and are difficult for cells to infiltrate and form new tissue. In this study, we developed a porous "click" organic-inorganic nanohybrid (PO-click-ON) cement that crosslinks via metal-free biorthogonal click chemistry and forms porous structures mimicking the native bone tissue via particulate leaching. Strain-promoted click reaction enables fast and efficient crosslinking of polymer chains with the exclusion of any toxic initiator or catalyst. The resulting PO-click-ON implants supported exceptional in vitro stem cell adhesion and osteogenic differentiation with a large portion of stem cells infiltrated deep into the scaffolds. In vivo study using a rat cranial defect model demonstrated that the PO-click-ON system achieved outstanding cell adsorption, neovascularization, and bone formation. The porous click cement developed in this study serves as a promising platform with multifunctionality for bone and other tissue engineering applications.

Single-level subject-specific finite element model can predict fracture outcomes in three-level spine segments under different loading rates.

Osteoporosis-related vertebral compression fracture can occur under normal physiological activities. Bone metastasis is another source of vertebral fracture. Different loading rates, either high-energy traumas such as falls or low-energy traumas under normal physiological activities, can result in different fracture outcomes. The aim of the current study was to develop a quantitative computed tomography-based finite element analysis (QCT/FEA) technique for single vertebral bodies to predict fracture strength of three-level spine segments. Developed QCT/FEA technique was also used to characterize vertebral elastic moduli at two loading rates of 5 mm/min, representing a physiologic loading condition, and 12000 mm/min, representing a high-energy trauma. To this end, a cohort of human spine segments divided into three groups of intact, defect, and augmented were mechanically tested to fracture; then, experimental stiffness and fracture strength values were measured. Outcomes of this study showed no significant difference between the elastic modulus equations at the two testing speeds. Areal bone mineral density measured by dual x-ray absorptiometry (DXA/BMD) explained only 53% variability (R2 = 0.53) in fracture strength outcomes. However, QCT/FEA could explain 70% of the variability (R2 = 0.70) in experimentally measured fracture strength values. Adding disk degeneration grading, testing speed, and sex to QCT/FEA-estimated fracture strength values further increased the performance of our statistical model by 14% (adjusted R2 of 0.84 between the prediction and experimental fracture forces). In summary, our results indicated that a single-vertebra model, which is computationally less expensive and more time efficient, is capable of estimating fracture outcomes with acceptable performance (range: 70-84%).

Injectable catalyst-free "click" organic-inorganic nanohybrid (click-ON) cement for minimally invasive in vivo bone repair.

Injectable polymers have attracted intensive attention in tissue engineering and drug delivery applications. Current injectable polymer systems often require free-radical or heavy-metal initiators and catalysts for the crosslinking process, which may be extremely toxic to the human body. Here, we report a novel polyhedral oligomeric silsesquioxane (POSS) based strain-promoted alkyne-azide cycloaddition (SPAAC) "click" organic-inorganic nanohybrids (click-ON) system that can be click-crosslinked without any toxic initiators or catalysts. The click-ON scaffolds supported excellent adhesion, proliferation, and osteogenesis of stem cells. In vivo evaluation using a rat cranial defect model showed outstanding bone formation with minimum cytotoxicity. Essential osteogenic alkaline phosphatase (ALP) and vascular CD31 marker expression were detected on the defect site, indicating excellent support of in vivo osteogenesis and vascularization. Using salt leaching techniques, an injectable porous click-ON cement was developed to create porous structures and support better in vivo bone regeneration. Beyond defect filling, the click-ON cement also showed promising application for spinal fusion using rabbits as a model. Compared to the current clinically used poly (methyl methacrylate) (PMMA) cement, this click-ON cement showed great advantages of low heat generation, better biocompatibility and biodegradability, and thus has great potential for bone and related tissue engineering applications.

Three-dimensional surface strain analyses of simulated defect and augmented spine segments: A biomechanical cadaveric study.

While several studies have investigated fracture outcomes of intact vertebrae, fracture properties in metastatically-involved and augmented vertebrae are still far from understood. Consequently, this study was aimed to use 3D digital image correlation (3D-DIC) method to investigate the failure properties of spine segments with simulated metastatic lesions, segments augmented with poly(propylene fumarate) (PPF), and compare the outcomes with intact spines. To this end, biomechanical experiments accompanied by 3D-DIC were performed on spine segments consisting of three vertebrae and two intervertebral discs (IVDs) at loading rates of 0.083 mm/s, mimicking a physiological loading condition, and 200 mm/s, mimicking an impact-type loading condition such as a fall or an accident. Full-field surface strain analysis indicated PPF augmentation reduces the superior/inferior strain when compared with the defect specimens; Presence of a defect in the middle vertebra resulted in shear band fracture pattern. Failure of the superior endplates was confirmed in several defect specimens as the superior IVDs were protruding out of defects. The augmenting PPF showed lower superior/inferior surface strain values at the fast speed as compared to the slow speed. The results of our study showed a significant increase in the fracture force from slow to fast speeds (p = 0.0246). The significance of the study was to determine the fracture properties of normal, pathological, and augmented spinal segments under physiologically-relevant loading conditions. Understanding failure properties associated with either defect (i.e., metastasis lesion) or augmented (i.e., post-treatment) spine segments could potentially provide new insights on the outcome prediction and treatment planning. Additionally, this study provides new knowledge on the effect of PPF augmentation in improving fracture properties, potentially decreasing the risk of fracture in osteoporotic and metastatic spines.

CT-based structural analyses of vertebral fractures with polymeric augmentation: A study of cadaveric three-level spine segments.

Pathologic vertebral fractures due to metastasis can occur under normal physiologic activities, leading to pain and neurologic deficit. Prophylactic vertebroplasty is a technique used to augment vertebral strength and reduce the risk of fracture. Currently, no technique is available to objectively assess vertebral fracture risk in metastatically-involved vertebral bodies. The aim of the current study was to develop an image-based computational technique to estimate fracture force outcomes during bending. To this end, mechanical testing was performed on intact, simulated defect, PMMA-augmented, and PPF-augmented 3-level spine segments from both sexes under a compression/flexion-type loading condition. The augmentation performance of poly(methyl methacrylate) (PMMA) and poly(propylene fumarate) (PPF) were also evaluated and compared. Cylindrical defects were created in 3-level spine segments with attached posterior elements and ligaments. Using CT images of each segment, a rigidity analysis technique was developed and used for predicting fracture forces during bending. On average, PPF strengthened the segments by about 630 N, resulting in fracture forces similar to those observed in the intact and PMMA-augmented groups. Female spines fractured at about 1150 N smaller force than did male spines. Rigidity analysis, along with age, explained 66% variability in experimental outcomes. This number increased to 74% when vertebral size and age were added to the rigidity analysis as explanatory variables. Both PPF and PMMA similarly increased fracture strength to the level of intact specimens. The results suggest that PPF can be a suitable candidate for augmentation purposes and rigidity analysis can be a promising predicting tool for vertebral fracture forces.

Density-Dependent Material and Failure Criteria Equations Highly Affect the Accuracy and Precision of QCT/FEA-Based Predictions of Osteoporotic Vertebral Fracture Properties.

About 700,000 vertebral fractures occur in the US as a result of bone loss. Quantitative computed tomography (QCT)-based finite element analysis (FEA) is a promising tool for fracture risk prediction that is becoming attractive in the clinical setting. The goals of this study were (1) to perform individual and pooled specimen optimization using inverse QCT/FEA modeling to obtain ash density-elastic modulus equations incorporating the whole vertebral body and accounting for all variables used during FE modeling, and (2) to determine the effect of material equations and failure criteria on the accuracy and precision of mechanical properties. Fifty-four (54) human vertebrae were used to optimize material equations based on experimental outcomes and, together with a previously proposed material equation, were implemented in our models using three different failure criteria to obtain fracture loads. Our robust QCT/FEA approach predicted 78% of the failure loads. Material equations resulted in poor accuracy in the predicted stiffness, yet yielded good precision and, more importantly, strong correlations with fracture loads. Both material and fracture criterion equations are equally important in estimating accurate and precise QCT/FEA predictions. Results suggest that both elastic modulus and fracture criterion equations should be validated against experimental outcomes to better explain the response of the tissue under various conditions.

Effect of metastatic lesion size and location on the load-bearing capacity of vertebrae using an optimized ash density-modulus equation.

About 1.8 million new cancer cases are estimated in the US in 2019 from which 50-85% might metastasize to the thoracic and lumbar spines. Subject-specific quantitative computed tomography-based finite element analysis (QCT/FEA) is a promising used tool to predict vertebral fracture properties. The aims of this study were twofold: First, to develop an optimized equation for the elastic modulus accounting for all input parameters in FE modeling of fracture properties. Second, to assess the effect of lesion size and location on the predicted fracture loads. An inverse QCT/FEA method was implemented to determine optimal coefficients for the modulus equation as a function of ash density. Lesions of 16 and 20 mm were then virtually located at the center, off-centered, anterior, and posterior regions of the vertebrae. A total of 6426 QCT/FEA models were run to optimize the coefficients and evaluate the effect of lesions on fracture properties. QCT/FEA predicted stiffness showed high correlations (50%) with the experimentally measured values. Compared to a 16 mm lesion size, a 20 mm lesion had a reduction in failure load of 55%, 57%, 52%, and 44% at the center, off-centered, anterior cortex, and pedicle, respectively (p < 0.001). Lesions affecting mostly trabecular bone showed the largest reduction in predicted failure loads (about 55%), and females presented weaker outcomes than males. An optimal elastic modulus equation resulted in accurate vertebral stiffness predictions. A deterioration of the trabecular bone due to the presence of a lesion highly affected the predicted fracture loads, and this reduction was significantly higher in females compared to males.

OPF/PMMA cage system as an alternative approach for the treatment of vertebral corpectomy.

The spinal column is the most common site for bone metastasis. Vertebral metastases with instability have historically been treated with corpectomy of the affected vertebral body and adjacent intervertebral discs, and are more recently treated with separation surgery. With demographics shifting towards an elderly population, a less invasive surgical approach is necessary for the repair of vertebral defects. We have modified a previously reported expandable hollow cage composed of an oligo[poly(ethylene glycol) fumarate] (OPF) containment system that could be delivered via a posterior-only approach. Then, the polymer of interest, poly(methyl methacrylate) (PMMA) bone cement, was injected into the lumen of the cage after expansion to form an OPF/PMMA cage. We compared six different cage formulations to account for vertebral body and defect size, and performed a cage characterization via expansion kinetics and mechanical testing evaluations. Additionally, we investigated the feasibility of the OPF/PMMA cage in providing spine stability via kinematic analyses. The in-vitro placement of the implant using our OPF/PMMA cage system showed improvement and mechanical stability in a flexion motion. The results demonstrated that the formulation and technique presented in the current study have the potential to improve surgical outcomes in minimally invasive procedures on the spine.

Mechanical testing setups affect spine segment fracture outcomes.

The purpose of the work presented here was to establish an experimental testing configuration that would generate a bending compression fracture in a laboratory setting. To this end, we designed and fabricated a fixture to accommodate a three level spine segment and to be able to perform mechanical testing by applying an off-centric compressive loading to create a flexion-type motion. Forces and moments occurring during testing were measured with a six-channel load cell. The initial testing configuration (Fixture A) included plates connected to the superior potted vertebral body and to the ball-socket joint of the testing system ram. Surprisingly, while all cadaveric specimens underwent a similar off-centric compressive loading, most of the specimens showed extension outcomes as opposed to the intended pure-flexion motion. The extension was due to fixture size and weight; by applying an off-centric load directly on the top plate, unintended large shear forces were generated. To resolve the issue, several modifications were made to the original fixture configuration. These modifications included the removal of the superior plates and the implementation of wedges at the superior surface of the fixture (Fixture B). A synthetic sample was used during this modification phase to minimize the number of cadaveric specimens while optimizing the process. The best outcomes were consistently observed when a 15°-wedge was used to provide flexion-type loading. Cadaveric specimens were then experimentally tested to fracture using the modified testing configuration (Fixture B). A comparison between both fixtures, A and B, revealed that almost all biomechanical parameters, including force, moment, and displacement data, were affected by the testing setup. These results suggest that fixture design and implementation for testing is of extreme importance, and can influence the fracture properties and affect the intended motion.

Optimizing Accuracy of Proximal Femur Elastic Modulus Equations.

Quantitative computed tomography-based finite element analysis (QCT/FEA) is a promising tool to predict femoral properties. One of the modeling parameters required as input for QCT/FEA is the elastic modulus, which varies with the location-dependent bone mineral density (ash density). The aim of this study was to develop optimized equations for the femoral elastic modulus. An inverse QCT/FEA method was employed, using an optimization process to minimize the error between the predicted femoral stiffness values and experimental values. We determined optimal coefficients of an elastic modulus equation that was a function of ash density only, and also optimal coefficients for several other equations that included along with ash density combinations of the variables sex and age. All of the optimized models were found to be more accurate than models from the literature. It was found that the addition of the variables sex and age to ash density made very minor improvements in stiffness predictions compared to the model with ash density alone. Even though the addition of age did not remarkably improve the statistical metrics, the effect of age was reflected in the elastic modulus equations as a decline of about 9% over a 60-year interval.

Factors associated with proximal femur fracture determined in a large cadaveric cohort.

Many researchers have used cadaveric fracture tests to determine the relationship between proximal femur (hip) fracture strength and a multitude of possible explanatory variables, typically considered one or two at a time. These variables include subject-specific proximal femur variables such as femoral neck areal bone mineral density (aBMD), sex, age, and geometry, as well as physiological hip fracture event variables such as fall speed and angle of impact. However, to our knowledge, no study has included all of these variables simultaneously in the same experimental dataset. To address this gap, the present study simultaneously included all of these subject-specific and fracture event variables in multivariate models to understand their contributions to femoral strength and fracture type. The primary aim of this study was to determine not only whether each of these variables contributed to the prediction of femoral strength, but also to determine the relative importance of each variable in strength prediction. A secondary aim was to similarly characterize the importance of these variables for the prediction of fracture type. To accomplish these aims, we characterized 197 proximal femurs (covering a wide range of subject-specific variables) with DXA and CT scans, and then tested the femurs to fracture in a sideways fall on the hip configuration. Each femur was tested using one of three fall speed conditions and one of four angles of impact (bone orientations). During each test, we acquired measurements of relevant force and displacement data. We then reduced the test data to determine femoral strength, and we used post-fracture CT scans to classify the fracture type (e.g., trochanteric, cervical). Using these results, the explanatory variables were analyzed with mixed statistical models to explain variations in hip fracture strength and fracture type, respectively. Five explanatory variables were statistically significant in explaining the variability in femoral strength: aBMD, sex, age, fall speed, and neck-shaft angle (P ≤ 0.0135). These five variables, including significant interactions, explained 80% of the variability in hip fracture strength. Additionally, when only aBMD, sex, and age (P < 0.0001) were considered in the model, again including significant interactions, these three variables alone explained 79% of the variability in hip fracture strength. So while fall speed (P = 0.0135) and neck-shaft angle (P = 0.0041) were statistically significant, the inclusion of these variables did not appreciably improve the prediction of hip fracture strength compared to the model that considered only aBMD, sex and age. For the variables we included in this study, in the ranges we considered, our findings indicate that the clinically-available information of patient age, sex and aBMD are sufficient for femoral strength assessment. These findings also suggest that there is little value in the extra effort required to characterize the effect of femoral geometry on strength, or to account for the probabilistic nature of fall-related factors such as fall speed and angle of impact. For fracture type, the only explanatory variable found to be significant was aBMD (P ≤ 0.0099). We found that the odds of having intertrochanteric fractures increased by 47% when aBMD decreased by one standard deviation (0.2 g/cm2).

Poly(Propylene Fumarate)-Hydroxyapatite Nanocomposite Can Be a Suitable Candidate for Cervical Cages.

A wide range of materials have been used for the development of intervertebral cages. Poly(propylene fumarate) (PPF) has been shown to be an excellent biomaterial with characteristics similar to trabecular bone. Hydroxyapatite (HA) has been shown to enhance biocompatibility and mechanical properties of PPF. The purpose of this study was to characterize the effect of PPF augmented with HA (PPF:HA) and evaluate the feasibility of this material for the development of cervical cages. PPF was synthesized and combined with HA at PPF:HA wt:wt ratios of 100:0, 80:20, 70:30, and 60:40. Molds were fabricated for testing PPF:HA bulk materials in compression, bending, tension, and hardness according to ASTM standards, and also for cage preparation. The cages were fabricated with and without holes and with porosity created by salt leaching. The samples as well as the cages were mechanically tested using a materials testing frame. All elastic moduli as well as the hardness increased significantly by adding HA to PPF (p < 0.0001). The 20 wt % HA increased the moduli significantly compared to pure PPF (p < 0.0001). Compressive stiffness of all cages also increased with the addition of HA. HA increased the failure load of the porous cages significantly (p = 0.0018) compared with nonporous cages. PPF:HA wt:wt ratio of 80:20 proved to be significantly stiffer and stronger than pure PPF. The current results suggest that this polymeric composite can be a suitable candidate material for intervertebral body cages.

Extensibility of the supraspinatus muscle can be predicted by combining shear wave elastography and magnetic resonance imaging-measured quantitative metrics of stiffness and volumetric fat infiltration: A cadaveric study.

BACKGROUND: A torn rotator cuff tendon will retract over time causing changes in muscle properties and decreasing its extensibility, or deformation. During surgery, large tensile loads are applied to bring the torn tendon to the footprint. Poor muscle extensibility and large tensile stresses at the repair might lead to gap formation or re-tear of the repair. A quantitative evaluation of muscle properties could be used to predict the extensibility of the supraspinatus (SSP) muscle. METHOD: Magnetic resonance imaging (MRI)-measured volumetric fat fraction and shear wave elastography (SWE)-measured elastic modulus of the SSP muscle were obtained on seventeen cadaveric shoulders. Experimental extensibility and stiffness were then measured by axially pulling the tendon up-to 60 N. Univariate and multivariate analyses were used to determine the correlation and contribution of fat fraction and elastic modulus to experimental outcomes. FINDINGS: SWE moduli negatively correlated with SSP muscle extensibility (r = 0.54-0.58, P ≤ 0.0259); fat fraction resulted in a positive correlation (r = 0.69, P = 0.0021). SWE measurements, solely, explained up to 34% and 33% of the variability in measured extensibility and stiffness, respectively. Fat Fraction, solely, explained 48% of the variability in extensibility and 36% of the variability in stiffness. These methods combined predicted up to 62% of the musculotendinous extensibility. INTERPRETATION: This study showed a comprehensive quantitative assessment of SSP muscle properties using SWE to estimate stiffness and MRI to measure fatty infiltration. The extensibility of the detached muscle/tendon unit was highly correlated to material properties of the muscle when these methods were used in combination.

The trabecular effect: A population-based longitudinal study on age and sex differences in bone mineral density and vertebral load bearing capacity.

BACKGROUND: Approximately 16-24% of postmenopausal women are affected by vertebral fractures, negatively affecting their quality of life. Trabecular and cortical bones in vertebrae decline differently with age, thus having a distinct impact on vertebral failure loads. The purpose of this study was to investigate the effect of trabecular and cortical volumetric bone mineral density loss over time on estimated failure loads; and to evaluate the effect of sex and age. METHOD: Fracture properties from a cohort of 82 patients were evaluated for L1-L3 vertebrae at baseline and 6th year using an image-based method that implements axial rigidity analysis. Cortical and trabecular volumetric bone mineral density were obtained, as well as their individual contribution to total failure load. Regression analyses were performed to determine the effect of age and sex on volumetric bone mineral density and failure loads. FINDINGS: Decline in trabecular and cortical volumetric bone mineral density, and failure load was sex-dependent (p ≤ 0.0095). Cortical and trabecular volumetric bone mineral density reduced 2.08 (g/cm3)/year and 2.02 (g/cm3)/year, respectively. A 1012 N difference in failure load, ~70% attributed to trabecular bone, was found between men and women of similar age. Over 6 years, this difference increased by 287 N. Areal bone mineral density measured by dual X-ray absorptiometry explained ~60% of the vertebral failure load. INTERPRETATION: Trabecular bone has a significantly greater effect than cortical bone on the structural integrity and load bearing capacity of vertebrae. This might lead to a higher incidence of fragility fractures in osteoporotic women. Our non-invasive, quantitative computed tomography image-based approach may improve prevention, monitoring, and management of fractures.

A Method to Estimate Cadaveric Femur Cortical Strains During Fracture Testing Using Digital Image Correlation.

This protocol describes the method using digital image correlation to estimate cortical strain from high speed video images of the cadaveric femoral surface obtained from mechanical testing. This optical method requires a texture of many contrasting fiduciary marks on a solid white background for accurate tracking of surface deformation as loading is applied to the specimen. Immediately prior to testing, the surface of interest in the camera view is painted with a water-based white primer and allowed to dry for several minutes. Then, a black paint is speckled carefully over the white background with special consideration for the even size and shape of the droplets. Illumination is carefully designed and set such that there is optimal contrast of these marks while minimizing reflections through the use of filters. Images were obtained through high speed video capture at up to 12,000 frames/s. The key images prior to and including the fracture event are extracted and deformations are estimated between successive frames in carefully sized interrogation windows over a specified region of interest. These deformations are then used to compute surface strain temporally during the fracture test. The strain data is very useful for identifying fracture initiation within the femur, and for eventual validation of proximal femur fracture strength models derived from Quantitative Computed Tomography-based Finite Element Analysis (QCT/FEA).

Method and Instrumented Fixture for Femoral Fracture Testing in a Sideways Fall-on-the-Hip Position.

Mechanical testing of femora brings valuable insights into understanding the contribution of clinically-measureable variables such as bone mineral density distribution and geometry on the femoral mechanical properties. Currently, there is no standard protocol for mechanical testing of such geometrically complex bones to measure strength, and stiffness. To address this gap we have developed a protocol to test cadaveric femora to fracture and to measure their biomechanical parameters. This protocol describes a set of adaptable fixtures to accommodate the various load magnitudes and directions accounting for possible bone orientations in a fall on the hip configuration, test speed, bone size, and left leg-right leg variations. The femora were prepared for testing by cleaning, cutting, scanning, and potting the distal end and greater trochanter contact surfaces in poly(methyl methacrylate) (PMMA) as presented in a different protocol. The prepared specimens were placed in the testing fixture in a position mimicking a sideways fall on the hip and loaded to fracture. During testing, two load cells measured vertical forces applied to the femoral head and greater trochanter, a six-axis load cell measured forces and moments at the distal femoral shaft, and a displacement sensor measured differential displacement between the femoral head and trochanter contact supports. High speed video cameras were used to synchronously record the sequence of fracture events during testing. The reduction of this data allowed us to characterize the strength, stiffness, and fracture energy for nearly 200 osteoporotic, osteopenic, and normal cadaveric femora for further development of engineering-based diagnostic tools for osteoporosis research.

Are DXA/aBMD and QCT/FEA Stiffness and Strength Estimates Sensitive to Sex and Age?

Dual X-ray absorptiometry (DXA) measures areal bone mineral density (aBMD) by simplifying a complex 3D bone structure to a 2D projection and is not equally effective for explaining fracture strength in women and men. Unlike DXA, subject-specific quantitative computed tomography-based finite element analysis (QCT/FEA) estimates fracture strength using 3D bone mineral distribution and geometry. By using experimentally-measured femoral stiffness and strength from a one hundred sample cadaveric cohort that included variations in sex and age, we wanted to determine if QCT/FEA estimates were able to better predict the experimental variations than DXA/aBMD. For each femur, DXA/aBMD was assessed and a QCT/FEA model was developed to estimate femoral stiffness and strength. Then, the femur was mechanically tested to fracture in a sideways fall on the hip position to measure stiffness and strength. DXA/aBMD and QCT/FEA estimates were compared for their sensitivity to sex and age with multivariate statistical analyses. When comparing the measured data with DXA/aBMD predictions, both age and sex were significant (p ≤ 0.0398) for both femoral stiffness and strength. However, QCT/FEA predictions of stiffness and strength showed sex was insignificant (p ≥ 0.23). Age was still significant (p ≤ 0.0072). These results indicate that QCT/FEA, unlike DXA/aBMD, accounted for bone differences due to sex.