Cortical Bone Specimens Research Articles

Experimental Analysis of Robotic Cortical Bone Specimen Drilling Performance: Effect of Cryogen.

In orthopedic surgery, precise bone screw insertion is crucial for stabilizing fractures, necessitating a preliminary cortical bone drilling procedure. However, this process can induce temperatures exceeding 70 °C due to the low thermal conductivity of cortical bone, potentially leading to thermal osteonecrosis. Furthermore, significant cutting forces and torque pose risks of tool breakage and bone damage, underlining the need for high precision and optimal processing parameters. Traditionally, drilling relies on the surgeon's experience and often results in imprecise outcomes due to inconsistent feed rates. Therefore, this study proposes the use of a 6-axis robot for controlled drilling, offering precise control over angular velocities and consistent feed rates. Additionally, explore the use of cryogenic liquid nitrogen (LN2) as a novel cooling method compared to conventional saline solutions, examining its efficacy under various cutting conditions. The results demonstrate that LN2 cooling conditions lead to a reduction in thrust and torque under specific processing conditions, and facilitate smoother chip evacuation. Additionally, LN2 significantly lowers the peak temperature around the drilling site, thus minimizing the risk of thermal osteonecrosis. Consequently, the use of a 6-axis robot provides consistent feed rates, and LN2 cooling achieves optimal processing conditions, enabling a more controlled and effective drilling process.

A novel quasi-static compression test set-up with micron order accuracy for small specimens

Accurate displacement measurements during compression tests on small specimens using large standard universal testing machines are adversely affected by the compliance of the apparatus. The influence of compliance becomes more acute when conducting tests on quasi-brittle materials, such as cortical bone, that exhibit low failure strains and for which accurate modulus values are required. This paper presents a custom compression test set-up (i.e., a subpress) that facilitates routine quasi-static compression tests on small specimens by eliminating the effect of testing machine compliance on the results. The displacement of the compression test set-up was recorded using a combination of Hall effect sensors and multipole magnetic strips, which offers a resolution of less than one micron. The results of quasi-static compression tests on small polymer specimens are reported where the displacements were measured using the custom compression test set-up, the integrated displacement measurement system of a universal testing machine and a visual extensometer. Additional tests on cortical bone specimens demonstrate the effectiveness of the compression test set-up. Based on the results, the compression test set-up appeared to yield more consistent and accurate measurements compared to both the integrated measurement system of a universal testing machine and a visual extensometer. Novel data regarding the incipient fracture of cortical bone were obtained. The compression test set-up allows for the routine testing of a large number of bone specimens with micron accuracy in a short time frame, thus reducing the effects of degradation on biological specimens.

Degradation of the mechanical properties of cortical bone due to long duration storage

Understanding the behaviour and material properties of bone is critical in predicting the failure and fracture of bones in humans. To address this, mechanical tests have traditionally been conducted to characterize bone material and this has resulted in large body of literature. However, there appears to be a lack of complete information regarding the storage protocols used for bone specimens prior to conducting mechanical tests. For example, while storage methods are well described, parameters such as the time between donor death and bone retrieval, as well as time between specimen machining and testing, are seldom reported. As biological materials undergo degradation in storage after being removed from the donor, a clear understanding of this degradation behaviour would identify critical time frames in which previously stored cortical bone specimens should be tested such that they can still be considered representative of an in-vivo condition. In this paper, the results of an investigation to determine the effects of long duration storage on the measured mechanical properties of bovine cortical bone are reported. Three different storage protocols are compared; namely machined-refrigerated, machined-frozen and frozen-machined-frozen. Degradation effects are evident for both refrigerated and frozen specimens and the results demonstrate that testing bone specimens after more than one week in storage may not provide representative in-vivo properties. In addition, specimens exhibit severe degradation after six months in storage regardless of the storage protocol.

An improved linear systems model of hydrothermal isometric tension testing to aid in assessing bone collagen quality: Effects of ribation and type-2 diabetes.

This study sought to further develop and validate a previously proposed physics-based model that maps denaturation kinetics from differential scanning calorimetry (DSC) to the isometric tension generated during hydrothermal isometric tension (HIT) testing of collagenous tissues. The primary objectives of this study were to verify and validate two physics-based model parameters: α, which indicates the amount of instantaneous isometric tension developed per unit of collagen denaturation, and β, which captures the proportionality between temperature and the generated isometric tension post denaturation initiation. These parameters were used as measures of bone collagen quality, employing data from HIT and DSC testing of human bone collagen from two previous studies. Additionally, given the physical basis of the model, the study aimed to further validate Max.Slope, the rate of change in isometric tensile stress with change in temperature, as an independent measure of collagen network connectivity. Max.Slope has previously been positively correlated with measures of cortical bone fracture resistance. Towards this verification and validation, the hypotheses were a) that α would correlate strongly with HIT denaturation temperature, Td, and the enthalpy of melting (ΔH) from DSC, and b) that β would correlate positively and strongly with Max.Slope.The model was employed in the analysis of HIT-DSC data from the testing of demineralized bone collagen isolated from cadaveric human femurs in two prior studies. In one study, data were collected from HIT-DSC testing of cortical bone collagen from 74 donors. Among them, 38 had a history of type 2 diabetes +/− chronic kidney disease, while the remaining 36 had no history of T2D again with or without CKD. Cortical bone specimens were extracted from the lateral mid-shaft. The second study involved 15 donor femora, with four cortical bone specimens extracted from each. Of these four, two specimens underwent a 4-week incubation in 0.1 M ribose at 37 °C to induce non-enzymatic ribation and advanced glycation endproducts, while the other two served as non-ribated controls.The examination involved investigating correlations between the model parameters α and β and various measures, such as Max.Slope, Td, ΔH, age, and duration of type 2 diabetes. The results revealed positive correlations between the model parameter β and Max.Slope (r = 0.55–0.58). The parameter α was found to be associated with Td, but also sensitive to the shape of the HIT curve around Td resulting in difficulties with variability and interpretation. As a result, while both hypotheses are confirmed, Max.Slope and β are better indicators of bone collagen quality because they are measures of the connectivity or, more generally, the integrity of the bone collagen network.

[Translated article] Mechanical properties variations of immature bone in the earliest stages of development

IntroductionBone as a material varies its composition and mechanical properties throughout life. Although these variations are better understood in adulthood, there is little experimental information on the variation of these properties in early stages of development. The objective of this study is to analyze the mechanical behavior and chemical properties of cortical bone tissue from two animal species in these earliest stages. Material and methodologyTwenty specimens of cortical bone were manufactured from bovine and ovine species that were in different stages of development (feeding exclusively on breast milk, in the transition period to feed or pasture, and young animals but on a solid food diet). The specimens were subjected to tensile tests, recorded with a high-speed camera to obtain deformation maps. Measurements of the tensile force until the specimen broke were also carried out. A fractographic study was carried out with a scanning electron microscope to analyze the fracture surface and an analysis of the amount of calcium in each of the specimens using X-ray dispersion spectroscopy. ResultsA statistically significant and positive correlation was found between the elastic modulus of the specimens and their calcium content. A trend towards more rigid behavior with age was observed. ConclusionsYoung bone tissue tends to stiffen with age as the calcium content increases with an increase in elastic modulus.

Osteoprogenitor recruitment and differentiation during intracortical bone remodeling of adolescent humans.

Recruitment and proliferation of osteoprogenitors during the reversal-resorption phase, and their differentiation into mature bone-forming osteoblasts is crucial for initiation of bone formation during bone remodeling. This study investigates the osteoprogenitors' gradual recruitment, proliferation, and differentiation into bone-forming osteoblasts within intracortical remodeling events of healthy adolescent humans. The study was conducted on cortical bone specimens from 11 adolescent human controls - patients undergoing surgery due to coxa valga. The osteoprogenitor recruitment route and differentiation into osteoblasts were backtracked using immunostainings and in situ hybridizations with osteoblastic markers (CD271/NGFR, osterix/SP7, COL3A1 and COL1A1). The osteoblastic cell populations were defined based on the pore surfaces, and their proliferation index (Ki67), density and number/circumference were estimated in multiplex-immunofluorescence (Ki67, TRAcP, CD34) stained sections. During the reversal-resorption phase, osteoclasts are intermixed with (COL3A1+NFGR+) osteoblastic reversal cells, which are considered to be osteoprogenitors of (COL1A1+SP7+) bone-forming osteoblasts. Initiation of bone formation requires a critical density of these osteoprogenitors (43±9 cells/mm), which is reached though proliferation (4.4±0.5% proliferative) and even more so through recruitment of osteoprogenitors, but challenged by the ongoing expansion of the canal circumference. These osteoprogenitors most likely originate from osteoblastic bone lining cells and mainly lumen osteoprogenitors, which expand their population though proliferation (4.6±0.3%) and vascular recruitment. These lumen osteoprogenitors resemble canopy cells above trabecular remodeling sites, and like canopy cells they extend above bone-forming osteoblasts where they may rejuvenate the osteoblast population during bone formation. Initiation of bone formation during intracortical remodeling requires a critical density of osteoprogenitors on eroded surfaces, which is reached though proliferation and recruitment of local osteoprogenitors: bone lining cells and lumen osteoprogenitors.

Variación de las propiedades mecánicas del hueso inmaduro en las etapas más tempranas del desarrollo

IntroductionBone as a material varies its composition and mechanical properties throughout life. Although these variations are better understood in adulthood, there is little experimental information on the variation of these properties in early stages of development. The objective of this study is to analyze the mechanical behavior and chemical properties of cortical bone tissue from two animal species in these earliest stages. Material and methodologyTwenty specimens of cortical bone were manufactured from bovine and ovine species that were in different stages of development (feeding exclusively on breast milk, in the transition period to feed or pasture, and young animals but on a solid food diet). The specimens were subjected to tensile tests, recorded with a high-speed camera to obtain deformation maps. Measurements of the tensile force until the specimen broke were also carried out. A fractographic study was carried out with a scanning electron microscope to analyze the fracture surface and an analysis of the amount of calcium in each of the specimens using X-ray dispersion spectroscopy. ResultsA statistically significant and positive correlation was found between the elastic modulus of the specimens and their calcium content. A trend towards more rigid behavior with age was observed. ConclusionsYoung bone tissue tends to stiffen with age as the calcium content increases with an increase in elastic modulus.

Evaluation of gadolinium deposition in cortical bone using three-dimensional ultrashort echo time quantitative susceptibility mapping: A preliminary study.

The aim of the current study was to investigate the feasibility of three-dimensional ultrashort echo time quantitative susceptibility mapping (3D UTE-QSM) for the assessment of gadolinium (Gd) deposition in cortical bone. To this end, 40 tibial bovine cortical bone specimens were divided into five groups then soaked in phosphate-buffered saline (PBS) solutions with five different Gd concentrations of 0, 0.4, 0.8, 1.2, and 1.6 mmol/L for 48 h. Additionally, eight rabbits were randomly allocated into three groups, consisting of a normal-dose macrocyclic gadolinium-based contrast agent (GBCA) group (n = 3), a high-dose macrocyclic GBCA group (n = 3), and a control group (n = 2). All bovine and rabbit tibial bone samples underwent magnetic resonance imaging (MRI) on a 3-T clinical MR system. A 3D UTE-Cones sequence was utilized to acquire images with five different echo times (i.e., 0.032, 0.2, 0.4, 0.8, and 1.2 ms). The UTE images were subsequently processed with the morphology-enabled dipole inversion algorithm to yield a susceptibility map. The average susceptibility was calculated in three regions of interest in the middle of each specimen, and the Pearson's correlation between the estimated susceptibility and Gd concentration was calculated. The bone samples soaked in PBS with higher Gd concentrations exhibited elevated susceptibility values. A mean susceptibility value of -2.47 ± 0.23 ppm was observed for bovine bone soaked in regular PBS, while the mean QSM value increased to -1.75 ± 0.24 ppm for bone soaked in PBS with the highest Gd concentration of 1.6 mmol/L. A strong positive correlation was observed between Gd concentrations and QSM values. The mean susceptibility values of rabbit tibial specimens in the control group, normal-dose GBCA group, and high-dose GBCA group were -4.11 ± 1.52, -3.85 ± 1.33, and -3.39 ± 1.35 ppm, respectively. In conclusion, a significant linear correlation between Gd in cortical bone and QSM values was observed. The preliminary results suggest that 3D UTE-QSM may provide sensitive noninvasive assessment of Gd deposition in cortical bone.

Approach to standardized material characterization of the human lumbopelvic system-Specification, preparation and storage.

The complexity of the osseo-ligamentous lumbopelvic system has made it difficult to perform both, the overall preparation as well as specimen harvesting and material testing with a reasonable amount of time and personnel. The logistics of such studies present a hurdle for reproducibility. A structured procedure was developed and proved, which allows all necessary steps to be carried out reproducibly and in a reasonable time. This enables the extraction of 26 soft tissue, 33 trabecular and 32 cortical bone specimens from this anatomical region per cadaver. The integrity of the specimens remains maintained while keeping requirements within manageable limits. The practicability of the intended five-day specimen harvesting and testing procedure could be demonstrated on five test and two pre-test sequences. The intended minimization of physical, biological, and chemical external influences on specimens could be achieved. All protocols, instructions and models of preparation and storage devices are included in the supporting information. The high grade of applicability and reproducibility will lead to better comparability between different biomechanical investigations. This procedure proven on the human pelvis is transferable to other anatomical regions.

A strain rate dependent model with decreasing Young’s Modulus for cortical human bone

In the existing literature, some studies have observed an increase in the elastic modulus of human cortical bone with strain rate, which has been described as a consequence of the viscoelastic properties of the bone. However, these results contradict the findings of other studies, in which an independence or decrease of the elastic modulus with strain rate is observed, which could be explained by other non-viscoelastic mechanisms. This research studies the dynamic behavior of human cortical bone specimens and investigates their mechanical properties . A full and objective strain rate dependent model is proposed and used to describe the experimental results obtained from uniaxial tensile tests of twenty-one human rib cortical bone specimens from twelve male post mortem human subjects (average age of 68.5 ± 12.3 years). In addition, a general discussion of some families of viscoelastic models is given and the caution with which they should be used when dealing with complex materials such as bone. The main experimental finding is that in the range of strain rate analyzed (), there is a significant decrease in Young’s modulus (E ≈ 18 GPa for and E ≈ 8 GPa for ), which is not of viscoelastic origin. Moreover, the most frequently used viscoelastic models analyzed in this study predict how the elastic modulus should not vary markedly with strain rate for small strains. In fact, the observed behavior seems related to the findings of other researchers who observed that the microcraking damage depends on the strain rate in the same sense found in our work. This allows us to interpret the qualitative results as a consequence of the microcracking that takes place within the cortical bone, and not related to viscoelastic effects.

An integrated experimental-computational framework to assess the influence of microstructure and material properties on fracture toughness in clinical specimens of human femoral cortical bone

Microstructural and compositional changes that occur due to aging, pathological conditions, or pharmacological treatments alter cortical bone fracture resistance. However, the relative importance of these changes to the fracture resistance of cortical bone has not been quantified in detail. In this technical note, we developed an integrated experimental-computational framework utilizing human femoral cortical bone biopsies to advance the understanding of how fracture resistance of cortical bone is modulated due to modifications in its microstructure and material properties. Four human biopsy samples from individuals with varying fragility fracture history and osteoporosis treatment status were converted to finite element models incorporating specimen-specific material properties and were analyzed using fracture mechanics-based modeling. The results showed that cement line density and osteonal volume had a significant effect on crack volume. The removal of cement lines substantially increased the crack volume in the osteons and interstitial bone, representing straight crack growth, compared to models with cement lines due to the lack of crack deflection in the models without cement lines. Crack volume in the osteons and interstitial bone increased when mean elastic modulus and ultimate strength increased and mean fracture toughness decreased. Crack volume in the osteons and interstitial bone was reduced when material property heterogeneity was incorporated in the models. Although both the microstructure and the heterogeneity of the material properties of the cortical bone independently increased the fracture toughness, the relative contribution of the microstructure was more significant. The integrated experimental-computational framework developed here can identify the most critical microscale features of cortical bone modulated by pathological processes or pharmacological treatments that drive changes in fracture resistance and improve our understanding of the relative influence of microstructure and material properties on fracture resistance of cortical bone.

Osteocyte apoptosis and cellular micropetrosis signify skeletal aging in type 1 diabetes.

Bone fragility is a profound complication of type 1 diabetes mellitus (T1DM), increasing patient morbidity. Within the mineralized bone matrix, osteocytes build a mechanosensitive network that orchestrates bone remodeling; thus, osteocyte viability is crucial for maintaining bone homeostasis. In human cortical bone specimens from individuals with T1DM, we found signs of accelerated osteocyte apoptosis and local mineralization of osteocyte lacunae (micropetrosis) compared with samples from age-matched controls. Such morphological changes were seen in the relatively young osteonal bone matrix on the periosteal side, and micropetrosis coincided with microdamage accumulation, implying that T1DM drives local skeletal aging and thereby impairs the biomechanical competence of the bone tissue. The consequent dysfunction of the osteocyte network hampers bone remodeling and decreases bone repair mechanisms, potentially contributing to the enhanced fracture risk seen in individuals with T1DM. STATEMENT OF SIGNIFICANCE: Type 1 diabetes mellitus (T1DM) is a chronic autoimmune disease that causes hyperglycemia. Increased bone fragility is one of the complications associated with T1DM. Our latest study on T1DM-affected human cortical bone identified the viability of osteocytes, the primary bone cells, as a potentially critical factor in T1DM-bone disease. We linked T1DM with increased osteocyte apoptosis and local accumulation of mineralized lacunar spaces and microdamage. Such structural changes in bone tissue suggest that T1DM speeds up the adverse effects of aging, leading to the premature death of osteocytes and potentially contributing to diabetes-related bone fragility.

Determination of the Modulus of Elasticity by Bending Tests of Specimens with Nonuniform Cross Section

BackgroundBending tests offer technical advantages when material testing is performed to determine the modulus of elasticity. In biomechanical studies, beam-like cortical bone specimens subjected to flexural loading are usually characterized by nonuniform cross-sectional properties along the beam axis and a comparatively large spatial variation of the local material properties.ObjectiveA suitable evaluation method for determining the average modulus of elasticity within the volume of beam-like specimens with nonuniform cross section was to be identified.MethodsA total of 138 samples of human pelvic cortical bone were extracted and tested under flexural loading. Different methods, all based on the linear-elastic flexural theory of beams, were applied to determine the average modulus of elasticity on the basis of measured deformations, and the results were compared. Some of these methods utilized the measured midspan deflection, and others used the elastic curve obtained by digital image correlation.ResultsThe results showed that it was not appropriate to determine the average modulus of elasticity from only the measured midspan deflection. The consideration of deflections at multiple points along the beam axis is recommended.ConclusionsAn evaluation method based on the fitting of the analytically determined elastic curve of the beam with its nonuniform cross-sectional properties to the measured deflections is considered the most appropriate method for determining the average modulus of elasticity of the specimen.

Mechanical properties variation of cortical bone with type of loading using FEM

Bone is known to be a natural composite material with variation of mechanical properties depending on the location and orientation of specimen extracted. In this study, specimens of cortical bone extracted from three different locations and two different orientations were modelled in Abaqus by considering the material of cortical bone to be anisotropic in nature. The stress–strain curves of specimens for both transverse and longitudinal orientation for compressive and tensile testing has been plotted. It has been found from the curves that the specimen belonging to upper diaphysis has more values of mechanical properties in comparison to other specimen-type for both compressive and tensile testing. It has also been observed that the compressive test shows more value of yield stress, elastic modulus and ultimate stress in comparison to tensile test that is the cortical bone can wear more load in the compressive direction. The values of mechanical properties found to be more for longitudinal orientation in comparison to transverse orientation. The results obtained found a good relation with the existing literature.

T1 relaxation of bound and pore water in cortical bone.

MRI measures of bound and/or pore water concentration in cortical bone offer potential diagnostics of bone fracture risk. The transverse relaxation characteristics of both bound and pore water are relatively well understood and have been used to design clinical MRI pulse sequences to image each water pool quantitatively. However, these methods are also sensitive to longitudinal relaxation characteristics, which have been less well studied. Here, spectroscopic relaxometry measurements of 31 human cortical bone specimens provided a more detailed picture of of both bound and pore water. The results included mean, standard deviation, and range of spectra from both bound and pore water, as well as novel presentations of the 2D distribution of pore water. Importantly, for each sample the pore water spectrum was found to span more than one order of magnitude and varied substantially across the 31 sample studies. Because many existing methods assume pore water to be mono-exponential and constant across individuals, the results were used to compute the potential effect neglecting this intra- and intersample variation on accurate MRI measurement of both bound and pore water concentrations. The greatest effect was found for adiabatic inversion recovery (AIR) based measurements of bound water concentration, which showed an average of 8.8% and as much as 37% error when using a common mono-exponential assumption of pore water . Despite these errors, the simulated AIR measurements were still moderately well correlated with the bound water concentrations derived from the spectroscopic data.

Accumulation of fluorescent advanced glycation end products and carboxymethyl-lysine in human cortical and trabecular bone

Chemical crosslinks known as advanced glycation end-products (AGEs) are associated with increased bone fracture risk and deteriorated bone mechanical properties. However, measurement of bone AGEs via ex vivo and in vitro methods has been limited to quantification of bulk fluorescent AGEs (fAGEs) and pentosidine only, which is a crosslinking fluorescent AGE. However, a non-crosslinking and non-fluorescent AGE such as carboxymethyl-lysine (CML) is found to be 40–100 times higher in quantity than pentosidine, but only one previous study has reported it in cortical bone, and one study reported it in trabecular bone. In our study, we wanted to investigate if accumulation of CML differs in cortical and trabecular compartments and if they are more strongly associated with bone mechanical properties than with fAGEs. We hypothesized that CML and fAGEs level would be higher in the trabecular compartment and show negative correlations to mechanical properties in cortical and trabecular bone. We obtained human cadaveric cortical and trabecular bone specimens, induced the formation of AGEs via the established in vitro ribosylation method, imaged specimens by microcomputed tomography to assess specimen geometry and microarchitecture, and mechanically tested cortical specimens by cyclic reference point indentation and fracture toughness tests and trabecular specimens by compression tests, followed by measurement of fAGEs and CML. fAGEs were 22 % higher in cortical bone (687 ± 44.8 ng Q/mg collagen) compared to trabecular bone (859 ± 317.1 ng Q/mg collagen), whereas CML levels were found to be 148 % higher in trabecular bone (6189.9 ± 866 ng/mg of protein) compared to cortical bone (924.6 ± 576.3 ng/mg of protein). Pooling the specimens from both the control and ribose groups, Spearman correlation analysis indicated that CML levels, but not fAGEs, are moderately associated with cortical porosity (r = +0.505, p ≤ 0.05) and mechanical properties such indentation depth (r = +0.460, p ≤ 0.05), total indentation depth (r = +0.440, p ≤ 0.05), and average energy dissipated (r = +0.465, p ≤ 0.05) in cortical bone. fAGEs showed a trend towards negative association with crack propagation toughness in cortical bone (r = −0.365, p = 0.055). No significant correlations were observed between CML and microarchitecture or mechanical properties in trabecular bone. CML levels were also associated with fAGEs in cortical bone (r = +0.596, p ≤ 0.05) but not in trabecular bone. Our preliminary findings indicate that CML, a non-crosslinking AGE, may affect bone material and mechanical properties differently than bulk fluorescent AGEs, given the higher accumulation of CML in each bone compartment. This study provides direction to future studies to quantify crosslinking and non-crosslinking AGEs separately as their effect on material and mechanical properties may be different and it would help identify better biomarkers for bone strength prediction.

The High-cycle Fatigue Life of Cortical Bone Allografts Is Radiation Sterilization Dose-dependent: An In Vitro Study.

Structural cortical bone allografts are a reasonable treatment option for patients with large cortical bone defects caused by trauma, tumors, or complications of arthroplasty. Although structural cortical bone allografts provide the benefit of an osteoconductive material, they are susceptible to fatigue failure (fracture) and carry a risk of disease transmission. Radiation-sterilization at the recommended dose of 25 kGy decreases the risk of disease transmission. However, previous studies demonstrated that radiation sterilization at this dose can negatively impact the high cycle-fatigue life of cortical bone. Although the effects of higher doses of radiation on cortical bone allografts are well described, the effects of lower doses of radiation on a high-cycle fatigue life of cortical bone are poorly understood. (1) Does the cycle-fatigue life of human cortical allograft bone vary with gamma radiation dose levels of 0 (control), 10 kGy, 17.5 kGy, and 25 kGy? (2) What differences in Raman spectral biomarkers are observed following varying doses of gamma radiation exposure? The high-cycle fatigue behavior of human cortical bone specimens was examined at different radiation sterilization doses under physiologic stress levels (35 MPa) and in a 37° C phosphate-buffered saline bath using a custom-designed rotating-bending fatigue device. Six human femora from three donors were obtained for this study (two male, 63 and 61 years old, respectively, and one female, 48 years old). Test specimens were allocated among four treatment groups (0 kGy [control], 10 kGy, 17.5 kGy, and 25 kGy) based on donor and anatomic location of harvest site (both length and cross-sectional quadrant of femoral diaphysis) to ensure equal variation (n = 13 per group). Specimens underwent high-cycle fatigue testing to failure. The number of cycles to failure was recorded. Raman spectroscopy (a noninvasive vibrational spectroscopy used to qualitatively assess bone quality) was used to detect whether any changes in Raman spectral biomarkers occurred after varying doses of gamma radiation exposure. There was a decrease in the log-transformed mean high-cycle fatigue life in specimens irradiated at 25 kGy (5.39 ± 0.32) compared with all other groups (0 kGy: 6.20 ± 0.50; 10k Gy: 6.35 ± 0.79; 17.5 kGy: 6.01 ± 0.53; p = 0.001). Specimens irradiated at 25 kGy were also more likely to exhibit a more brittle fracture surface pattern than specimens with more ductile fracture surface patterns irradiated at 0 kGy, 10 kGy, and 17.5 kGy (p = 0.04). The Raman biomarker for the ratio of the relative amount of disordered collagen to ordered collagen showed a decrease at the 10 kGy radiation level from 1.522 ± 0.025 preirradiation to 1.489 ± 0.024 postirradiation (p = 0.01); no other detectable changes in Raman biomarkers were observed. The high-cycle fatigue life of cortical bone undergoes a nonlinear, dose-dependent decrease with an increase in gamma radiation sterilization in a clinically relevant dose range (0-25 kGy). Importantly, a notable drop-off in the high-cycle fatigue life of cortical bone appeared to occur between 17.5 kGy and 25 kGy, correlating to a sixfold decrease in mean cycles to failure. We speculate that the decrease in the Raman biomarker for disordered collagen at 10 kGy with no loss in high-cycle fatigue life may be caused by an increased amount of nonenzymatic crosslinking of the collagen backbone relative to collagen chain-scission (whereas the benefits of crosslinking may be outweighed by excess scission of the collagen backbone at higher radiation doses), but future studies will need to ascertain whether this in fact is the case. Radiation sterilization at the industry standard of 25 kGy has a substantial negative impact on the high-cycle fatigue life of cortical bone. Given these findings, it is possible to provide a meaningful increase in the high-cycle fatigue life and improve the overall functional lifetime of cortical bone allografts by lowering the radiation-sterilization dose below 25 kGy. Future work on radiation-sterilization methods at these clinically relevant doses is warranted to aid in preserving the high cycle fatigue life of cortical bone allografts while maintaining sterility.

Inverse Problem in Resonant Ultrasound Spectroscopy With Sampling and Optimization: A Comparative Study on Human Cortical Bone.

The Bayesian inference with prior knowledge has been proposed recently to solve the inverse problem in resonant ultrasound spectroscopy. It allows inferring the elastic properties of high damping materials, such as cortical bone with less dependence on the initial guessed values. In this method, the estimation of the stiffness coefficients is expressed as a probabilistic solution to the inverse problem, which can be achieved by sampling or optimization methods. However, the detailed performance comparison of these two strategies applied to high damping materials has not been fully studied. In this work, the full stiffness tensor of 52 transversely isotropic cortical bone specimens was obtained using Markov chain Monte Carlo (MCMC) sampling and particle swarm optimization (PSO), respectively. Results showed that the local probability distributions of stiffness coefficients estimated by the two methods are consistent. Compared with MCMC, the average calculation speed of PSO is ten times faster [614 s ± 59 s (MCMC) versus 53 s ± 22 s (PSO)]. The mean standard error between theoretical and experimental resonant frequencies was slightly smaller for PSO compared with MCMC. In conclusion, PSO, a global optimization strategy, is suitable to solve the inverse problem for high damping materials.

An in vitro bone-to-bone adhesion test method using the compression shear test

The development of new bone adhesives is severely restricted due to the lack of a standard test method to examine their bond strength. Therefore, this study presents an in vitro bone-to-bone adhesion test method based on a compression shear test using cortical bone specimens (35 × 15 × 3 mm). The zero-gap lap joint test configuration was used to measure the bonding strengths of two soft tissue adhesives (Histoacryl® and BSA-Glue) and a bone cement (Palacos® LV). For reproducible bonding results, it is crucial to work with specimen having reproducible surface, dimensions and where and how they were taken from a bone. Therefore, this is a focus of the work. The adhesive strengths were determined under dry and physiological conditions to evaluate the reliability of the test procedure. Furthermore, the influence of previous acid etching of the bone surface was also investigated. The adhesives achieved bond strengths of 5.5 MPa (Histoacryl®), 3.2 MPa (BSA-Glue) and 2.8 MPa (Palacos® LV) under dry conditions. Under physiological conditions, the measured adhesive strengths of all three bonding materials decreased by around 90%. The acid etching led to a higher presence of reactive amino groups on the bone surface and improved the adhesion of Histoacryl® and BSA-Glue. The Palacos® LV bone cement suffered a reduction in adhesion as the acid smoothened the bone surface, resulting in less mechanical interlocking. The proposed test method is based on known standards and enables the usability of medical adhesives as a bone adhesive to be evaluated in a reproducible manner.

Influence of X-rays and gamma-rays on the mechanical performance of human bone factoring out intraindividual bone structure and composition indices.

Doses of irradiation above 25 ​kGy are known to cause irreversible mechanical decay in bone tissue. However, the impact of irradiation doses absorbed in a clinical setting on the mechanical properties of bone remains unclear. In daily clinical practice and research, patients and specimens are exposed to irradiation due to diagnostic imaging tools, with doses ranging from milligray to Gray. The aim of this study was to investigate the influence of irradiation at these doses ranges on the mechanical performance of bone independent of inter-individual bone quality indices.Therefore, cortical bone specimens (n ​= ​10 per group) from a selected organ donor were irradiated at doses of milligray, Gray and kilogray (graft tissue sterilization) at five different irradiation doses. Three-point bending was performed to assess mechanical properties in the study groups.Our results show a severe reduction in mechanical performance (work to fracture: 50.29 ​± ​11.49 Nmm in control, 14.73 ​± ​1.84 Nmm at 31.2 ​kGy p ​≤ ​0.05) at high irradiation doses of 31.2 ​kGy, which correspond to graft tissue sterilization or synchrotron imaging. In contrast, no reduction in mechanical properties were detected for doses below 30 ​Gy. These findings are further supported by fracture surface texture imaging (i.e. more brittle fracture textures above 31.2 ​kGy).Our findings show that high radiation doses (≥31.2 ​kGy) severely alter the mechanical properties of bone. Thus, irradiation of this order of magnitude should be taken into account when mechanical analyses are planned after irradiation. However, doses of 30 ​Gy and below, which are common for clinical and experimental imaging (e.g., radiation therapy, DVT imaging, CT imaging, HR-pQCT imaging, DXA measurements, etc.), do not alter the mechanical bending-behavior of bone.