MicroCT study on the enamel mineral density of primary molars.

AIM: The aim of this study is to report on the mineral density of the enamel of primary molars related to the age of the child and to compare the mineral density of sound and carious enamel in those molars. MATERIALS AND METHODS: This study included 23 children and 41 extracted primary molars. The primary molars of 21 children met all of the inclusion criteria, and these were studied and scanned using microCT. The teeth were embedded in Impregum (3M ESPE) and stored in a solution of tap water with thymol crystals. Sixteen primary molars from 7 children were used to compare the mineral density in sound and carious areas, and 13 primary molars from 11 children were used for the comparison between mineral density and time in situ. RESULTS: A statistically significant difference (31%) was found between the mineral density in carious enamel and sound enamel (p = 0.0006). In addition, a significant relationship was observed between the mineral density of sound enamel and the time the teeth had been in situ (r = 0.698). We also found two teeth with radiolucencies in the dentin with the enamel clinically showing only a non-cavitated carious lesion in the enamel. No significant differences were found between the mean mineral density in sound enamel surfaces and unaffected areas in surfaces of molars with enamel caries (p = 0.4373). CONCLUSION: Local and general differences in enamel mineralisation are presented. Post-eruptive maturation seems to be present not only in permanent teeth but also in primary molars. Carious enamel has significantly less mineral density than clinically sound enamel.

Demineralized bone matrix and platelet-rich plasma do not improve healing of osteochondral defects of the talus: an experimental goat study.

OBJECTIVE: The purpose of this study was to evaluate the effectiveness of demineralized bone matrix (DBM) with and without platelet-rich plasma (PRP) in the treatment of osteochondral defects (OCDs) of the talus. We hypothesized that treatment with DBM would result in more bone formation than no treatment in control OCDs, and that PRP would further enhance the regenerative capacity of DBM. METHOD: A standardized 6-mm OCD was created in each talus of 16 adult goats. According to a randomization scheme, one OCD of each goat was treated with allogeneic DBM hydrated with normal saline (n = 8) or hydrated with autologous PRP (n = 8). The contralateral OCD (n = 16) served as control. After 24 weeks, the animals were euthanized and the tali excised. Various outcome parameters were analyzed with use of macroscopic evaluation, micro-computed tomography (µCT), histology, histomorphometry, and fluorescence microscopy. RESULTS: None of the analyses revealed statistically significant differences between the groups for any of the parameters analyzed in any volume of interest. For example, the mean bone volume fraction (BV/TV) of the defect, as measured by µCT, was 0.56 (95% confidence interval [CI], 0.44-0.68) for DBM hydrated with normal saline and 0.52 (95% CI, 0.40-0.65) for DBM hydrated with PRP, compared to 0.53 (95% CI, 0.45-0.61) and 0.54 (95% CI, 0.44-0.64) for the internal controls, respectively (P > 0.05). CONCLUSION: In contrast to our hypotheses, no beneficial treatment effect of DBM with or without PRP was found for OCDs of the caprine talus.

Biomechanical effect of mineral heterogeneity in trabecular bone.

Due to daily loading, trabecular bone is subjected to deformations (i.e., strain), which lead to stress in the bone tissue. When stress and/or strain deviate from the normal range, the remodeling process leads to adaptation of the bone architecture and its degree of mineralization to effectively withstand the sustained altered loading. As the apparent mechanical properties of bone are assumed to depend on the degree and distribution of mineralization, the goal of the present study was examine the influences of mineral heterogeneity on the biomechanical properties of trabecular bone in the human mandibular condyle. For this purpose nine right condyles from human dentate mandibles were scanned and evaluated with a microCT system. Cubic regional volumes of interest were defined, and each was transformed into two different types of finite element (FE) models, one homogeneous and one heterogeneous. In the heterogeneous models the element tissue moduli were scaled to the local degree of mineralization, which was determined using microCT. Compression and shear tests were simulated to determine the apparent elastic moduli in both model types. The incorporation of mineralization variation decreased the apparent Young's and shear moduli by maximally 21% in comparison to the homogeneous models. The heterogeneous model apparent moduli correlated significantly with bone volume fraction and degree of mineralization. It was concluded that disregarding mineral heterogeneity may lead to considerable overestimation of apparent elastic moduli in FE models.

Spatial orientation in bone samples and Young's modulus.

Bone mass is the most important determinant of the mechanical strength of bones, and spatial structure is the second. In general, the spatial structure and mechanical properties of bones such as the breaking strength are direction dependent. The mean intercept length (MIL) and line frequency deviation (LFD) are two methods for quantifying directional aspects of the spatial structure of bone. Young's modulus is commonly used to describe the stiffness of bone, which is also a direction-dependent mechanical property. The aim of this article is to investigate the relation between MIL and LFD on one hand and Young's modulus on the other. From 11 human mandibular condyles, 44 samples were taken and scanned with high-resolution computer tomography equipment (micro-CT). For each sample the MIL and LFD were determined in 72602 directions distributed evenly in 3D space. In the same directions Young's modulus was determined by means of the stiffness tensor that had been determined for each sample by finite element analysis. To investigate the relation between the MIL and LFD on one hand and Young's modulus on the other, multiple regression was used. On average the MIL accounted for 69% of the variance in Young's modulus in the 44 samples and the LFD accounted for 72%. The average percentage of variance accounted for increased to 80% when the MIL was combined with the LFD to predict Young's modulus. Obviously MIL and LFD to some extent are complementary with respect to predicting Young's modulus. It is known that directional plots of the MIL tend to be ellipses or ellipsoids. It is speculated that ellipsoids are not always sufficient to describe Young's modulus of a bone sample and that the LFD partly compensates for this.

Regional variations in mineralization and strain distributions in the cortex of the human mandibular condyle.

The strain (i.e. deformation) history influences the degree of mineralization of cortical bone (DMB) as well as its osteonal microstructure. This study aimed to examine the relationships of stress and strain distributions with the variations in DMB and the osteonal orientations in the cortical bone of the human mandibular condyle. It was hypothesized that strains are inversely proportional to local DMB and that the principal strains are oriented parallel to the osteons. To test this, ten human mandibular condyles were scanned in a microCT system. Finite element models were created in order to simulate static clenching. Within each condyle, 18 volumes of interest were selected to analyze regional differences in DMB, stress and strains. Subchondral bone showed a lower equivalent strain (2652+/-612 muepsilon) as compared to the anterior (p=0.030) and posterior cortex (p=0.007) and was less mineralized. Contrary to our hypothesis, the results show that strains correlated positively with regional variations in DMB (r=0.750, p<0.001). In the anterior and the posterior cortex, the first principal strain was parallel to the cortical surface and oriented supero-inferiorly with a fan-like shape. In subchondral bone, the first and the second principal strain were parallel to the surface and oriented antero-posteriorly and medio-laterally, respectively. It was concluded that the strain distributions, by themselves, cannot explain the regional differences found in DMB. In agreement with our second hypothesis, the orientation of the osteonal network of the mandibular condyle was closely related to the strain orientations. The results of this study suggest that the subchondral and the cortical bone are structured to ensure an optimal load distribution within the mandibular condyle and have a different mechanical behaviour. Subchondral bone plays a major role in the transmission of the strains to the anterior and posterior cortex, while these ensure an optimal transmission of the strains within the condylar neck and, eventually, to the mandibular ramus.

Variations in mineralization affect the stress and strain distributions in cortical and trabecular bone.

The mechanical properties of bone depend largely on its degree and distribution of mineralization. The present study analyzes the effect of an inhomogeneous distribution of mineralization on the stress and strain distributions in the human mandibular condyle during static clenching. A condyle was scanned with a micro-CT scanner to create a finite element model. For every voxel the degree of mineralization (DMB) was determined from the micro-CT scan. The Young's moduli of the elements were calculated from the DMB using constant, linear, and cubic relations, respectively. Stresses, strains, and displacements in cortical and trabecular bone, as well as the condylar deformation (extension along the antero-posterion axis) and compliance were compared. Over 90% of the bone mineral was located in the cortical bone. The DMB showed large variations in both cortical bone (mean: 884, SD: 111 mg/cm(3)) and trabecular bone (mean: 738, SD: 101 mg/cm(3)). Variations of the stresses and the strains were small in cortical bone, but large in trabecular bone. In the cortical bone an inhomogeneous mineral distribution increased the stresses and the strains. In the trabecular bone, however, it decreased the stresses and increased the strains. Furthermore, the condylar compliance remained relatively constant, but the condylar deformation doubled. It was concluded that neglect of the inhomogeneity of the mineral distribution results in a large underestimation of the stresses and strains of possibly more than 50%. The stiffness of trabecular bone strongly influences the condylar deformation. Vice versa, the condylar deformation largely determines the magnitude of the strains in the trabecular bone.

Structural and mechanical properties of mandibular condylar bone.

The trabecular bone of the mandibular condyle is structurally anisotropic and heterogeneous. We hypothesized that its apparent elastic moduli are also anisotropic and heterogeneous, and depend on trabecular density and orientation. Eleven condyles were scanned with a micro-CT system. Volumes of interest were selected for the construction of finite element models. We simulated compressive and shear tests to determine the principal mechanical directions and the apparent elastic moduli. Compressive moduli were relatively large in directions acting in the sagittal plane, and small in the mediolateral direction. The degree of mechanical anisotropy ranged from 4.7 to 10.8. Shear moduli were largest in the sagittal plane and smallest in the transverse plane. The magnitudes of the moduli varied with the condylar region and were proportional to the bone volume fraction. Furthermore, principal mechanical direction correlated significantly with principal structural direction. It was concluded that variation in trabecular structure coincides with variation in apparent mechanical properties.

Long-term vitamin D deficiency in older adult C57BL/6 mice does not affect bone structure, remodeling and mineralization.

Animal models show that vitamin D deficiency may have severe consequences for skeletal health. However, most studies have been performed in young rodents for a relatively short period, while in older adult rodents the effects of long-term vitamin D deficiency on skeletal health have not been extensively studied. Therefore, the first aim of this study was to determine the effects of long-term vitamin D deficiency on bone structure, remodeling and mineralization in bones from older adult mice. The second aim was to determine the effects of long-term vitamin D deficiency on mRNA levels of genes involved in vitamin D metabolism in bones from older adult mice. Ten months old male C57BL/6 mice were fed a diet containing 0.5% calcium, 0.2% phosphate and 0 (n=8) or 1 (n=9) IU vitamin D3/gram for 14 months. At an age of 24 months, mice were sacrificed for histomorphometric and micro-computed tomography (micro-CT) analysis of humeri as well as analysis of CYP27B1, CYP24 and VDR mRNA levels in tibiae and kidneys using RT-qPCR. Plasma samples, obtained at 17 and 24 months of age, were used for measurements of 25-hydroxyvitamin D (25(OH)D) (all samples), phosphate and parathyroid hormone (PTH) (terminal samples) concentrations. At the age of 17 and 24 months, mean plasma 25(OH)D concentrations were below the detection limit (<4nmol/L) in mice receiving vitamin D deficient diets. Plasma phosphate and PTH concentrations did not differ between both groups. Micro-CT and histomorphometric analysis of bone mineral density, structure and remodeling did not reveal differences between control and vitamin D deficient mice. Long-term vitamin D deficiency did also not affect CYP27B1 mRNA levels in tibiae, while CYP24 mRNA levels in tibiae were below the detection threshold in both groups. VDR mRNA levels in tibiae from vitamin D deficient mice were 0.7 fold lower than those in control mice. In conclusion, long-term vitamin D deficiency in older adult C57BL/6 mice, accompanied by normal plasma PTH and phosphate concentrations, does not affect bone structure, remodeling and mineralization. In bone, expression levels of CYP27B1 are also not affected by long-term vitamin D deficiency in older adult C57BL/6 mice. Our results suggest that mice at old age have a low or absent response to vitamin D deficiency probably due to factors such as a decreased bone formation rate or a reduced response of bone cells to 25(OH)D and 1,25(OH)2D. Older adult mice may therefore be less useful for the study of the effects of vitamin D deficiency on bone health in older people.

The effect of bone loss on rod-like and plate-like trabeculae in the cancellous bone of the mandibular condyle.

Bone loss may affect the structure of cancellous bone. But its effect on trabeculae with different characteristics, like rods and plates, is not accurately known. This study analyzes the effect of bone loss on individual rod-like and plate-like trabeculae. 94 specimens were obtained from mandibular condyles from both dentate and edentate humans and scanned with a micro-CT scanner. The bone volume fraction (BV/TV) of these specimens ranged from 7% to 30%. Next, the rod-like and plate-like trabeculae were identified with an especially developed algorithm. Plate volume fraction (PV/TV), rod volume fraction (RV/TV), plate thickness, rod thickness, number of plates, and number of rods were determined. In individual specimens, the thickness of the rods ranged from 40 mum to 180 mum, while the thickness of the plates ranged from 40 mum to 300 mum. In every specimen, the thickness of the plates was larger than the thickness of the rods. Statistical analysis revealed that PV/TV was proportional (r = 0.98, P < 0.001) and RV/TV inversely proportional (r = -0.45, P < 0.001) to BV/TV. Also the thickness of the plates correlated with BV/TV (r = 0.62, P < 0.001), while the thickness of the rods (mean = 90 mum, SD = 7 mum) remained constant (r = -0.09, P = 0.378). A four-fold reduction of the bone volume fraction was accompanied by a three-fold reduction of the number of plates and a 40% reduction of their thickness, but also by a three-fold increase in the number of rods and RV/TV. It was concluded that the effect of bone loss on plate-like trabeculae was opposite to its effect on rod-like trabeculae. Remarkably, the thickness of the rods (90 mum) was independent of the bone volume fraction. This suggests that there is a minimal thickness for trabeculae.

A telemetry system to chronically record muscle activity in middle-sized animals.

Radio-telemetry enables the long-term recordings of biopotentials that may be obtained in freely moving animals without interference by the experimenter. The purpose of this study was to test a fully implantable device for: (1) its transmission range; (2) the characteristics of the transmitted signals; and (3) its actual application in long-term in vivo registration of EMG. Transmission range was tested by changing the device's position relative to the receiver. Computer simulation of the filtering characteristics provided comparison of original and transmitted signals. Implantation of the device in masticatory muscles, followed by analysis of telemetred signals and determination of activity levels allowed for examination of daily muscle use. The implant's transmission range covered the cage size for middle-sized animals with a minimum of signal dropouts. Transmitted signals were marked by (partial) loss of frequencies beyond 50 Hz, decreased amplitude and slightly delayed timing relative to original waveforms. Analysis of the transmitted EMG revealed that the device can be used for prolonged in vivo EMG registration, detection of peak activity levels, and the examination of general muscle use by the time spent at different levels of activity.

Prediction of mechanical properties of the cancellous bone of the mandibular condyle.

The mechanical properties of cancellous bone depend on the bone structure. The present study examined the extent to which the apparent stiffness of the cancellous bone of the human mandibular condyle can be predicted from its structure. Two models were compared. The first, a structure model, used structural parameters such as bone volume fraction and anisotropy to estimate the apparent stiffness. The second was a finite element model (FEM) of the cancellous bone. The bone structure was characterized by micro-computed tomography. The calculated stiffnesses of 24 bone samples were compared with measured stiffnesses. Both models could predict 89% of the variation in the measured stiffnesses. From the stiffness approximated by FEM in combination with the measured stiffness, the stiffness of the bone tissue was estimated to be 11.1 +/- 3.2 GPa. It was concluded that both models could predict the stiffness of cancellous bone with adequate accuracy.

Three-dimensional finite element analysis of the cartilaginous structures in the human temporomandibular joint.

While the movability of the human temporomandibular joint is great, the strains and stresses in the cartilaginous structures might largely depend on the position of the mandible with respect to the skull. This hypothesis was investigated by means of static three-dimensional finite element simulations involving different habitual condylar positions. Furthermore, the influence of several model parameters was examined by sensitivity analyses. The results indicated that the disc moved together with the condyle in the anterior direction without the presence of ligaments and the lateral pterygoid muscle. By adapting its shape to the changing geometry of the articular surfaces, the disc prevented small contact areas and thus local peak loading. In a jaw-closed configuration, the influence of 30 degrees variations of the loading direction was negligible. The load distribution capability of the disc appeared to be proportional to its elasticity and was enhanced by the fibrocartilage layers on the articular surfaces.

Mechanical significance of the trabecular microstructure of the human mandibular condyle.

The human mandibular condyle has a parasagittal plate-like trabecular structure. We tested the hypothesis that this structure reflects the mechanical loading of the condyle. We developed a finite element model of the condyle to analyze the strains occurring during static compressive loading. The principal strains in the trabecular bone were primarily oriented in the sagittal plane. The first component was compressive and oriented supero-inferiorly. The second component was negligibly small and oriented medio-laterally. The third component was tensile, oriented antero-posteriorly, and almost equal to the compressive strain. This tensile strain was caused by antero-posterior bulging of the cortex. This means that the trabecular structure is also subjected to significant tensile forces. The orientation of the parasagittal strains followed the direction of the applied load. It was concluded that the trabecular structure of the mandibular condyle is optimal in resisting the compressive and tensile strains to which it is subjected.

Electromyographic heterogeneity in the human temporalis and masseter muscles during dynamic tasks guided by visual feedback.

The complex architecture of the human jaw muscles suggests regional differences in function within these muscles. This study examines the way the temporalis and masseter muscle regions are activated when free mandibular movements with various speeds and against various external leads are carried out guided by visual feedback. Electromyographic (EMG) activity was registered in six temporalis and three masseter muscle regions with bipolar fine-wire electrodes. Recordings were made during open/close excursions, protrusion/retrusion movements, and laterodeviations. During open/close excursions and protrusion/retrusion movements, an anterior and posterior temporalis part could be distinguished, whereas during laterodeviations a more complex partitioning of this muscle was observed. During the protrusion/retrusion movements and the laterodeviations, the temporalis muscle demonstrated higher EMG peak activities than the masseter muscle, and within the masseter muscle the deep masseter showed higher EMG peaks than the superficial one. In contrast to this, during the open/close excursions the masseter showed higher peak activities than the temporalis muscle, while the superficial masseter showed higher EMG peak activities than the deep masseter. Within the deep masseter, differences were also found. During open/close excursions, the anterior deep region demonstrated higher EMG peak activities than the posterior region, whereas during protrusion/retrusion and laterodeviations the posterior deep region showed higher peaks. In general, speed had a greater effect on the EMG peak activity than external load. Only during laterodeviations did speed and load equally influence peak activity in both the deep and superficial masseter. During protrusion/retrusion movements, load showed no significant effect on EMG peak activity in the masseter muscle. A general finding was that, according to task, different regions were activated preferentially. This points to a partitioning of the excitatory command of the motoneuron pool.

The influence of specimen attachment and dimension on microtensile strength.

The higher microtensile bond strength values found for specimens with a smaller cross-sectional area are often explained by the lower occurrence of internal defects and surface flaws. We hypothesized that this aberrant behavior is mainly caused by the lateral way of attachment of the specimens to the testing device, which makes the strength dependent on the thickness. This study showed that composite bars of 1x1x10, 1x2x10, and 1x3x10mm attached at their 1-mm-wide side (situation A) fractured at loads of the same magnitude, as a result of which the microtensile strength ( micro TS), calculated as F/A (force at fracture/cross-sectional area), significantly increased for specimens with decreasing thickness. Attachment at the 1-, 2-, or 3-mm-wide side (situation B) resulted in equal micro TS values (P > 0.05). Finite element analysis showed different stress patterns for situation A, but comparable patterns for situation B. Both situations showed the same maximum stress at fracture.

Fitting parametrized polynomials with scattered surface data.

Currently used joint-surface models require the measurements to be structured according to a grid. With the currently available tracking devices a large quantity of unstructured surface points can be measured in a relatively short time. In this paper a method is presented to fit polynomial functions to three-dimensional unstructured data points. To test the method spherical, cylindrical, parabolic, hyperbolic, exponential, logarithmic, and sellar surfaces with different undulations were used. The resulting polynomials were compared with the original shapes. The results show that even complex joint surfaces can be modelled with polynomial functions. In addition, the influence of noise and the number of data points was also analyzed. From a surface (diam: 20 mm) which is measured with a precision of 0.2 mm a model can be constructed with a precision of 0.02 mm.

The accuracy of joint surface models constructed from data obtained with an electromagnetic tracking device.

Electromagnetic tracking devices are widely used in biomechanics. In this article a method is evaluated to construct models of articular surfaces using an electromagnetic tracking device. First, the accuracy of the space tracker was examined and optimised. Then, from several joint surfaces random points were measured and eighth degree polynomials were fitted to these measurements. To check if the fit converged well, plots of cross sections of the model with corresponding data points were examined. The accuracy of the models was determined by comparing them with computed tomography data and by reproducibility tests. All the fits converged well to the data. The root mean square (RMS) error of the models varied from 0.07 to 0. 18mm, and was proportional to the size and complexity of the surface. This was mainly due to systematic errors made by the space tracker, which were also proportional to the size and complexity of the surface.

Passive force characteristics of an architecturally complex muscle.

In architecturally complex muscles with large attachment areas, it can be expected that during movement different muscle regions undergo different amounts of length excursions. As a consequence, the amount of passive force produced by the regions will differ. Therefore, we tested the hypothesis that during movement the vector of the passive force of such a muscle, which defines the magnitude, position and orientation of the resultant force of the various regions, has no fixed position, between the muscle's center of origin and insertion. As a model for an architecturally complex muscle we used the masseter muscle. It was expected that during jaw opening anterior muscle regions are more stretched than posterior regions, leading to an anterior shift of the passive force vector. A three-component force transducer was used to measure both the position and magnitude of passive force in the masseter muscle of 9 rabbits. Forces were recorded during repeated cycles of stepwise opening and closure of the jaw. The muscle exhibited a clear hysteresis: passive force measured during jaw opening was larger than that during jaw closing. With an increase of the jaw gape there was an approximately exponential increase of the magnitude of the passive muscle force, while simultaneously the passive force vector shifted anteriorly. Moment arm length of passive force increased by about 100%. This anterior shift contributed substantially to the increase of the passive muscle moment generated during jaw opening. It can be concluded that in architecturally complex muscles the increase of the passive resistance moment which is associated with muscle lengthening might not only be due to an increase of the magnitude of passive muscle force but also to an increase of the moment arm of this force.

A new transducer system for direct motor unit force measurement.

A new transducer was developed for in situ measurement of the force vector in a complex muscle. The transducer measures the magnitude, and the line of action of a force in a single plane. The dynamic range of the transducer is 0-5 N. This range includes the small forces developed by an active motor unit and the relatively large passive force of a whole muscle. In this study we present the details of the transducer design and specifications, and describe its application in the measurement of motor unit forces of the rabbit masseter muscle.

Three-dimensional finite element analysis of the human temporomandibular joint disc.

A three-dimensional finite element model of the articular disc of the human temporomandibular joint has been developed. The geometry of the articular cartilage and articular disc surfaces in the joint was measured using a magnetic tracking device. First, polynomial functions were fitted through the coordinates of these scattered measurements. Next, the polynomial description was transformed into a triangulated description to allow application of an automatic mesher. Finally, a finite element mesh of the articular disc was created by filling the geometry with tetrahedral elements. The articulating surfaces of the mandible and skull were modeled by quadrilateral patches. The finite element mesh and the patches were combined to create a three-dimensional model in which unrestricted sliding of the disc between the articulating surfaces was allowed. Simulation of statical joint loading at the closed jaw position predicted that the stress and strain distributions were located primarily in the intermediate zone of the articular disc with the highest values in the lateral part. Furthermore, it was predicted that considerable deformations occurred for relatively small joint loads and that relatively large variations in the direction of joint loading had little influence on the distribution of the deformations.