Influence of Bone-Level Dental Implants Placement and of Cortical Thickness on Osseointegration: In Silico and In Vivo Analyses.

The purpose of this research is to study the biomechanical response of dental implants in bone-level type locations, 0.5 mm above and below the bone level. In addition, the influence of the thickness of the cortical bone on osseointegration is determined due to the mechanical loads transfer from the dental implant to the cortical and trabecular bone. The thicknesses studied were 1.5 mm and 2.5 mm. Numerical simulations were performed using a finite element method (FEM)-based model. In order to verify the FEM model, the in silico results were compared with the results obtained from a histological analysis performed in an in vivo study with 30 New Zealand rabbits. FEM was performed using a computerized 3D model of bone-level dental implants inserted in the lower jawbone with an applied axial load of 100 N. The analysis was performed using different distances from the bone level and different thicknesses of cortical bone. The interface area of bone growth was evaluated by analyzing the bone-implant contact (BIC), region of interest (ROI) and total bone area (BAT) parameters obtained through an in vivo histological process and analyzed by scanning electron microscopy (SEM). Bone-level implants were inserted in the rabbit tibiae, with two implants placed per tibia. These parameters were evaluated after three or six weeks of implantation. FEM studies showed that placements 0.5 mm below the bone level presented lower values of stress distribution compared to the other studied placements. The lower levels of mechanical stress were then correlated with the in vivo studies, showing that this position presented the highest BIC value after three or six weeks of implantation. In this placement, vertical bone growth could be observed up the bone level. The smallest thickness of the study showed a better transfer of mechanical loads, which leads to a better osseointegration. In silico and in vivo results both concluded that the implants placed 0.5 mm below the cortical bone and with lower thicknesses presented the best biomechanical and histological behavior in terms of new bone formation, enhanced mechanical stability and optimum osseointegration.

Distal skeletal tibia assessed by HR-pQCT is highly correlated with femoral and lumbar vertebra failure loads.

Dual energy X-ray absorptiometry (DXA) is the standard for assessing fragility fracture risk using areal bone mineral density (aBMD), but only explains 60-70% of the variation in bone strength. High-resolution peripheral quantitative computed tomography (HR-pQCT) provides 3D-measures of bone microarchitecture and volumetric bone mineral density (vBMD), but only at the wrist and ankle. Finite element (FE) models can estimate bone strength with 86-95% precision. The purpose of this study is to determine how well vBMD and FE bone strength at the wrist and ankle relate to fracture strength at the hip and spine, and to compare these relationships with DXA measured directly at those axial sites. Cadaveric samples (radius, tibia, femur and L4 vertebra) were compared within the same body. The radius and tibia specimens were assessed using HR-pQCT to determine vBMD and FE failure load. aBMD from DXA was measured at the femur and L4 vertebra. The femur and L4 vertebra specimens were biomechanically tested to determine failure load. aBMD measures of the axial skeletal sites strongly correlated with the biomechanical strength for the L4 vertebra (r=0.77) and proximal femur (r=0.89). The radius correlated significantly with biomechanical strength of the L4 vertebra for vBMD (r=0.85) and FE-derived strength (r=0.72), but not with femur strength. vBMD at the tibia correlated significantly with femoral biomechanical strength (r=0.74) and FE-estimated strength (r=0.83), and vertebral biomechanical strength for vBMD (r=0.97) and FE-estimated strength (r=0.91). The higher correlations at the tibia compared to radius are likely due to the tibia's weight-bearing function.

Mechanical stimuli of trabecular bone in osteoporosis: A numerical simulation by finite element analysis of microarchitecture.

Mechanical stimuli are one of the factors that influence bone cell activity and therefore the remodeling of bone. These stimuli are dependent on the microarchitecture of the tissue and can be altered by changes in the bone that occur typically with osteoporosis. The objective of this study was to quantify the variation in the mechanical stimuli of trabecular bone due to changes in the microarchitecture. The morphology of 76 cubes of trabecular bone from human tibia were obtained from microcomputed tomography images and estimated possibilities for mechanical stimuli were determined using poro-viscoelastic finite element models based on the three-dimensional images. The distributions of Von Mises stress, octahedral strain, strain energy density, fluid velocity and pore pressure were predicted for the solid and the marrow phases of bone. We predicted that with variations in the morphology of the trabecular bone, such as an increase of 30% porosity, there is a significant decrease in the mechanical stimuli of the tissue when subjected to constant strain. The average stress and strain in the bone phase may reduce 50% and the fluid velocity in the marrow phase 88%. These decreases may intrinsically affect the mechanoregulation of bone regeneration that contributes to the etiology of osteoporosis.

Human trabecular bone microarchitecture can be assessed independently of density with second generation HR-pQCT.

The second generation HR-pQCT scanner (XtremeCTII, Scanco Medical) can assess human bone microarchitecture of peripheral limbs with a 61 µm nominal isotropic voxel size. This is a marked improvement from the first generation HR-pQCT that had a nominal isotropic voxel size of 82 µm, which is at the limit to accurately determine the thickness of individual human trabeculae. We sought to determine the accuracy of a direct morphometric approach to measure trabecular bone microarchitecture with three-dimensional morphological techniques using second generation HR-pQCT, and to compare this with the approach currently applied by the first generation HR-pQCT scanner based on derived indices using ex vivo scans of human cadaveric radii. We also compared images acquired and resampled to mimic the first generation HR-pQCT with those obtained directly from the first generation HR-pQCT. We evaluated 20 human cadaveric radii and a micro-CT performance phantom using the first (XtremeCT, Scanco Medical) and second generation HR-pQCT scanner (XtremeCTII) and compared a patient evaluation (XCTII, 61 µm) with a high resolution ex vivo protocol (HR, 30µm). We generated 82 µm scans of the same specimens to mimic a first-generation HR-pQCT evaluation (XCTIM, 82 µm) and compared these with a first-generation patient evaluation (XCTI, 82 µm). A standard structural extraction approach was applied to both XCTII and HR evaluations for assessment of bone volume fraction (BV/TV), and a distance transform was used to assess trabecular number (Tb.N), trabecular thickness (Tb.Th) and trabecular separation (Tb.Sp). For XCTI and XCTIM evaluations we followed the manufacturer's standard procedure and assessed bone mineral density (BMD), Tb.N with a distance transform, and then derived bone volume ratio (BV/TV(d)), trabecular thickness (Tb.Th(d)) and separation (Tb.Sp(d)). The spatial resolution (10% MTF) was 142.2 µm for XCTI, 108.9 µm for XCTIM, 95.2µm for XCTII, and 55.9 µm for HR. XCTI and XCTIM provided strongly associated measurements of BMD and microarchitectural outcomes (R(2)>0.97), however there were systematic differences in all outcomes. The Tb.N was highly associated with HR by both XCTII (R(2)=0.93, mean error=-0.12 mm(-1)) and XCTIM (R(2)=0.98, mean error=0.25 mm(-1)). Also, both XCTII (R(2)=0.99, mean error=0.20mm) and XCTIM (R(2)=0.99, mean error=-0.18 mm) had Tb.Sp that were strongly related to HR. For Tb.Th, the XCTII was more closely related to HR (R(2)=0.94, mean error=0.04 mm) than the relatively weak XCTIM (R(2)=0.16, mean error=- 0.076 mm). We found that trabecular microarchitecture assessment following the XCTII direct morphometric approach accurately represented the HR data. In particular, the measure of Tb.Th was markedly improved for XCTII compared with the derived approach of XCTIM. These data support the application of analysis techniques in HR-pQCT that are analogous to those traditionally used for micro-CT to assess trabecular microarchitecture. The decreased dependence of structural outcomes on density provides a new, important opportunity to monitor human in vivo bone microarchitecture.

The poro-viscoelastic properties of trabecular bone: a micro computed tomography-based finite element study.

Bone is a porous structure with a solid phase that contains hydroxyapatite and collagen. Due to its composition, bone is often represented either as a poroelastic or as a viscoelastic material; however, the poro-viscoelastic formulation that allows integrating the effect of both the fluid flow and the collagen on the mechanical response of the tissue, has not been applied yet. The objective of this study was to develop a micro computed tomography (µCT)-based finite element (FE) model of trabecular bone that includes both the poroelastic and the viscoelastic nature of the tissue. Cubes of trabecular bone (N=25) from human distal tibia were scanned with µCT and stress relaxation experiments were conducted. The µCT images were the basis for sample specific FE models, and the stress relaxation experiments were simulated applying a poro-viscoelastic formulation. The model considers two scales of the tissue: the intertrabecular pore and the lacunar-canalicular pore scales. Independent viscoelastic and poroelastic models were also developed to determine their contribution to the poro-viscoelastic model. All the experiments exhibited a similar relaxation trend. The average reaction force before relaxation was 9.28 × 10(2)N (SD ± 5.11 × 10(2)N), and after relaxation was 4.69 × 10(2)N (SD ± 2.88 × 10(2)N). The slope of the regression line between the force before and after relaxation was 1.92 (R(2)=0.96). The poro-viscoelastic models captured 49% of the variability of the experimental data before relaxation and 33% after relaxation. The relaxation predicted with viscoelastic models was similar to the poro-viscoelastic ones; however, the poroelastic formulation underestimated the reaction force before relaxation. These data suggest that the contribution of viscoelasticity (fluid flow-independent mechanism) to the mechanical response of the tissue is significantly greater than the contribution of the poroelasticity (fluid flow-dependent mechanism).

A comparison of methods for in vivo assessment of cortical porosity in the human appendicular skeleton.

The recent advent of high-resolution peripheral quantitative computed tomography (HR-pQCT) provides new opportunities to measure in vivo human bone microarchitecture. Increasingly, cortical porosity (CtPo) is of particular interest due to its relationship with bone quality and turnover. The two approaches that have emerged to measure CtPo from HR-pQCT are threshold-based and density-based methods, and the purpose of this work was to compare the performance of each against a gold-standard synchrotron radiation micro-computed tomography (SRµCT) measurement. Human cadaveric cortical bone specimens (N=23) were measured by SRµCT and HR-pQCT, and high correlations were found for both methods. The density-based approach had an r2=0.939 (95% confidence interval (CI) of +6.17% to +20.99%) and consistently overestimated porosity as measured by SRµCT, while the threshold-based approach had an r2=0.977 and consistently underestimated porosity (95% CI of -2.60% to -10.76%). The density-based approach is prone to beam hardening artifacts and susceptible to natural variations of tissue mineral density (TMD), but is less affected by motion artifacts that may occur in in vivo scans. The threshold-based method has the advantage that it provides structural information that complements the cortical porosity measure, such as number of pores and connectivity, and can accurately detect the larger pores which are the most relevant to bone biomechanical strength. With the first generation HR-pQCT systems the accuracy of detecting pores larger than 140 µm diameter is excellent (r2=0.983; 95% CI of -4.88% to +2.45%). The accuracy of the threshold-based method will improve as new HR-pQCT systems emerge and provide a robust quantitative approach to measure cortical porosity.

Predicting the permeability of trabecular bone by micro-computed tomography and finite element modeling.

The intrinsic permeability of bone plays an important role in the transport of nutrients and minerals within the tissue, and affects the mechanical stimuli that are related to the fate of the stem cells. The objective of this study was to establish a method to assess trabecular bone permeability using experimental and finite element (FE) modeling approaches based on micro computed tomography (µCT) images. Human cadaveric tibia cube specimens (N=23) were scanned with µCT. The permeability was measured experimentally using a custom-developed constant-head permeameter, and computationally by a poroelastic formulation to simulate the fluid flow within the discretized bone matrix and pore phase. The average of the experimentally measured permeability was 4.84 × 10(-10)m(2) with a standard deviation of 3.70 × 10(-10)m(2). A regression model of the µCT determined that the maximum bone area to total area ratio (maxBA/TA) for all slices that are perpendicular to the direction of fluid flow explained 84% of the variability of the natural logarithm of the experimentally measured permeability. The 2D measure of maxBA/TA performed better than 3D measures in general, although some parameters were reasonably well associated with permeability such as bone volume ratio (BV/TV, r=-0.71), the bone surface/bone volume (BS/BV, r=0.73), and the trabecular thickness (TbTh, r=-0.71). The correlation between the permeability predicted with FE models and experimentally measured permeability was reasonable (r=0.69), but the FE approach did not accurately represent the wide variability of permeability measured experimentally. The results of this study suggest that the changes in the trabecular bone microarchitecture have an exponential relationship with permeability, and the use of µCT-based 2D measurement of maxBA/TA performs well at predicting permeability, thus providing a convenient approach to measure this important aspect affecting biomechanical functions in the tissue.

A dynamical study of the mechanical stimuli and tissue differentiation within a CaP scaffold based on micro-CT finite element models.

The control of the mechanical stimuli transmitted to the cells is critical for the design of functional scaffolds for tissue engineering. The objective of this study was to investigate the dynamics of the mechanical stimuli transmitted to the cells during tissue differentiation in an irregular morphology scaffold under compressive load and perfusion flow. A calcium phosphate-based glass porous scaffold was used. The solid phase and the fluid flow within the pores were modeled as linear elastic solid material and Newtonian fluid, respectively. In the fluid model, different levels of viscosity were used to simulate tissue differentiation. Compressive strain of 0.5% and fluid flow with constant inlet velocity of 10 µm/s or constant inlet pressure of 3 Pa were applied. Octahedral shear strain and fluid shear stress were used as mechano-regulatory stimuli. For constant inlet velocity, stimuli equivalent to bone were predicted in 80% of pore volume for the case of low tissue viscosity. For the cases of high viscosity, fluctuations between stimuli equivalent to tissue formation and cell death were predicted due to the increase in the fluid shear stress when tissue started to fill pores. When constant pressure was applied, stimuli equivalent to bone were predicted in 62% of pore volume when low tissue viscosity was used and 42% when high tissue viscosity was used. This study predicted critical variations of fluid shear stress when cells differentiated. If these variations are not controlled in vitro, they can impede the formation of new matured tissue.

Simulation of angiogenesis and cell differentiation in a CaP scaffold subjected to compressive strains using a lattice modeling approach.

Mechanical stimuli are one of the factors that influence tissue differentiation. In the development of biomaterials for bone tissue engineering, mechanical stimuli and formation of a vascular network that transport oxygen to cells within the pores of the scaffolds are essential. Angiogenesis and cell differentiation have been simulated in scaffolds of regular porosity; however, the dynamics of differentiation can be different when the porosity is not uniform. The objective of this study was to investigate the effect of the mechanical stimuli and the capillary network formation on cell differentiation within a scaffold of irregular morphology. A porous scaffold of calcium phosphate based glass was used. The pores and the solid phase were discretized using micro computed tomography images. Cell activity was simulated within the interconnected pore domain of the scaffold using a lattice modeling approach. Compressive strains of 0.5 and 1% of total deformation were applied and two cases of mesenchymal stem cells initialization (in vitro seeding and in vivo) were simulated. Similar capillary networks were formed independently of the cell initialization mode and the magnitude of the mechanical strain applied. Most of vessels grew in the pores at the periphery of the scaffolds and were blocked by the walls of the scaffold. When 0.5% of strain was applied, 70% of the pore volume was affected by mechano-regulatory stimuli corresponding to bone formation; however, because of the lack of oxygen, only 40% of the volume was filled with osteoblasts. 40% of volume was filled with chondrocytes and 3% with fibroblasts. When the mechanical strain was increased to 1%, 11% of the pore volume was filled with osteoblasts, 59% with chondrocytes, and 8% with fibroblasts. This study has shown the dynamics of the correlation between mechanical load, angiogenesis and tissue differentiation within a scaffold with irregular morphology.