[Design of new gradient scaffolds based on triply periodic minimal surfaces and study on its mechanical, permeability and tissue differentiation characteristics].

In order to establish a bone scaffold with good biological properties, two kinds of new gradient triply periodic minimal surfaces (TPMS) scaffolds, i.e., two-way linear gradient G scaffolds (L-G) and D, G fusion scaffold (N-G) were designed based on the gyroid (G) and diamond (D)-type TPMS in this study. The structural mechanical parameters of the two kinds of scaffolds were obtained through the compressive simulation. The flow property parameters were also obtained through the computational fluid dynamics (CFD) simulation in this study, and the permeability of the two kinds of scaffolds were calculated by Darcy's law. The tissue differentiation areas of the two kinds of scaffolds were calculated based on the tissue differentiation theory. The results show that L-G scaffold has a better mechanical property than the N-G scaffold. However, N-G scaffold is better than the L-G scaffold in biological properties such as permeability and cartilage differentiation areas. The modeling processes of L-G and N-G scaffolds provide a new insight for the design of bone scaffold. The simulation in this study can also give reference for the prediction of osseointegration after the implantation of scaffold in the human body.

Effects of treadmill with different intensities on bone quality and muscle properties in adult rats.

BACKGROUND: Bone is a dynamically hierarchical material that can be divided into length scales of several orders of magnitude. Exercise can cause bone deformation, which in turn affects bone mass and structure. This study aimed to study the effects of treadmill running with different intensities on the long bone integrity and muscle biomechanical properties of adult male rats. METHODS: Forty-eight 5-month-old male SD rats were randomly divided into 4 groups: i.e., sedentary group (SED), exercise with speed of 12 m/min group (EX12), 16 m/min group (EX16), and 20 m/min group (EX20). The exercise was carried out for 30 min every day, 5 days a week for 4 weeks. The femurs were examined using three-point bending test, microcomputer tomography scanning and nanoindentation test; the soleus muscle was dissected for tensile test; ALP and TRACP concentrations were measured by serum analysis. RESULTS: The failure load was significantly increased by the EX12 group, whereas the elastic modulus was not significantly changed. The microstructure and mineral densities of the trabecular and cortical bone were significantly improved by the EX12 group. The mechanical properties of the soleus muscle were significantly increased by treadmill exercise. Bone formation showed significant increase by the EX12 group. Statistically higher nanomechanical properties of cortical bone were detected in the EX12 group. CONCLUSION: The speed of 12 m/min resulted in significant changes in the microstructure and biomechanical properties of bone; besides, it significantly increased the ultimate load of the soleus muscle. The different intensities of treadmill running in this study provide an experimental basis for the selection of exercise intensity for adult male rats.

Multiscale experimental study on the effects of different weight-bearing levels during moderate treadmill exercise on bone quality in growing female rats.

BACKGROUND: Bone tissue displays a hierarchical organization. Mechanical environments influence bone mass and structure. This study aimed to explore the effects of different mechanical stimuli on growing bone properties at macro-micro-nano scales. METHODS: Sixty five-week-old female Wistar rats were treadmill exercised at moderate intensity with the speed of 12 m/min, and then randomly divided into five groups according to weight-bearing level. After 8 weeks of experiment, femurs were harvested to perform multiscale tests. RESULTS: Bone formation was significantly increased by weight-bearing exercise, whereas bone resorption was not significantly inhibited. Trabecular and cortical bone mineral densities showed no significant increase by weight-bearing exercise. The microstructure of trabecular bone was significantly improved by 12% weight-bearing exercise. However, similar positive effects were not observed with further increase in weight-bearing levels. The nanomechanical properties of trabecular bone were not significantly changed by weight-bearing exercise. The macrostrength of whole femur and the nanomechanical properties of cortical bone significantly decreased in the 19% and 26% weight-bearing exercise groups. CONCLUSION: When rats ran on the treadmill at moderate intensity during growth period, additional 12% weight-bearing level could significantly increase bone formation, improve microstructure of trabecular bone, as well as maintain the structure and mechanical properties of cortical bone. Excessive weight-bearing level caused no positive effects on the trabecular bone microstructure and properties of cortical bone at all scales. In addition, increased weight-bearing level exerted no significant influence on trabecular and cortical bone mineral densities.

Multiscale investigation on the effects of additional weight bearing in combination with low-magnitude high-frequency vibration on bone quality of growing female rats.

This study aimed to explore the effects of additional weight bearing in combination with low-magnitude high-frequency vibration (LMHFV; 45 Hz, 0.3 g) on bone quality. One hundred twenty rats were randomly divided into ten groups; namely, sedentary (SED), additional weight bearing in which the rat wears a backpack whose weight is x% of the body weight (WBx; x = 5, 12, 19, 26), basic vibration (V), and additional weight bearing in combination with LMHFV in which the rat wears a backpack whose weight is x% of the body weight (Vx; x = 5, 12, 19, 26). The experiment was conducted for 12 weeks, 7 days per week, and 15 min per day. A three-point bending mechanical test, micro computed tomography, and a nanoindentation test were used. Serum samples were analyzed chemically. Failure load in V19 rats was significantly lower than that in SED rats (P < 0.05). Vx (x = 5, 12, 19, 26) rats showed poor microarchitectures. The content of tartrate-resistant acid phosphatase 5b was significantly higher in Vx (x = 5, 12, 19, 26) rats than that in SED rats (P < 0.05). V26 rats demonstrated comparatively better nanomechanical properties of materials than the other vibrational groups. Additional weight bearing in combination with LMHFV negatively affected the macromechanical properties and microarchitecture of bone. Heavy additional weight bearing, such as 26% of body weight, in combination with LMHFV was able to improve the nanomechanical properties of growing bone material compared with LMHFV. A combined mechanical stimulation was used, which may provide useful information to understand the mechanism of this mechanical stimulation on bone.

Quantification of Age-Related Tissue-Level Failure Strains of Rat Femoral Cortical Bones Using an Approach Combining Macrocompressive Test and Microfinite Element Analysis.

Bone mechanical properties vary with age; meanwhile, a close relationship exists among bone mechanical properties at different levels. Therefore, conducting multilevel analyses for bone structures with different ages are necessary to elucidate the effects of aging on bone mechanical properties at different levels. In this study, an approach that combined microfinite element (micro-FE) analysis and macrocompressive test was established to simulate the failure of male rat femoral cortical bone. Micro-FE analyses were primarily performed for rat cortical bones with different ages to simulate their failure processes under compressive load. Tissue-level failure strains in tension and compression of these cortical bones were then back-calculated by fitting the experimental stress-strain curves. Thus, tissue-level failure strains of rat femoral cortical bones with different ages were quantified. The tissue-level failure strain exhibited a biphasic behavior with age: in the period of skeletal maturity (1-7 months of age), the failure strain gradually increased; when the rat exceeded 7 months of age, the failure strain sharply decreased. In the period of skeletal maturity, both the macro- and tissue-levels mechanical properties showed a large promotion. In the period of skeletal aging (9-15 months of age), the tissue-level mechanical properties sharply deteriorated; however, the macromechanical properties only slightly deteriorated. The age-related changes in tissue-level failure strain were revealed through the analysis of male rat femoral cortical bones with different ages, which provided a theoretical basis to understand the relationship between rat cortical bone mechanical properties at macro- and tissue-levels and decrease of bone strength with age.

Spatiotemporal Characterization of Microstructure Morphology, Mechanical Properties and Bone Remodeling of Rat Tibia Under Uniaxial Compressive Overload Loading.

Bone tissue is subjected to increased mechanical stress during high-intensity work. Inadequate bone remodeling reparability can result in the continuous accumulation of microdamage, leading to stress fractures. The aim of this work was to investigate the characteristics and repair mechanisms of tibial microdamage under several degrees of overload. Also, we aimed at better understanding the effects of overload on the multi-scale structure and mechanical properties of bone. Sixty 5-month female rats were divided into three groups with different time points. Micro-CT was used to evaluate the three-dimensional microstructure, and three-point bending, quasi-static fracture toughness and creep mechanical test were carried out to evaluate the mechanical properties. SEM was used to observe the morphological characteristics of fracture surfaces. Section staining was used to count the microdamage parameters and numbers of osteoblasts and osteoclasts. The microarchitectures of cancellous and cortical bones in the three overload groups showed different degrees of damage. Overload led to a messy crystal structure of cortical bone, with slender microcracks mixed in, and a large number of broken fibers of cancellous bone. The properties associated with the elastic plasticity, fracture toughness, and viscoelasticity of cortical bone reduced in three groups, with that corresponding to day 30 presenting the highest damage. The accumulation of microdamage mainly occurred in the first 14 days, that is, the crack density peaked on day 14. Peak-targeted bone remodeling of cortical and cancellous bones occurred mainly between days 14 and 30. The influence of overload mechanical environment on bone quality at different time points was deeply investigated, which is of great significance for the etiology and treatment of stress fractures.

Relationship between microstructure, material distribution, and mechanical properties of sheep tibia during fracture healing process.

The aim of this study was to investigate the relationship between microstructural parameters, material distribution, and mechanical properties of sheep tibia at the apparent and tissue levels during the fracture healing process. Eighteen sheep underwent tibial osteotomy and were sacrificed at 4, 8, and 12 weeks. Radiographs and micro-computed tomography (micro-CT) scanning were taken for microstructural assessment, material distribution evaluation, and micro-finite element analysis. A displacement of 5% compressive strain on the longitudinal direction was applied to the micro-finite element model, and apparent and tissue-level mechanical properties were calculated. Principle component analysis and linear regression were used to establish the relationship between principle components (PCs) and mechanical parameters. Visible bony callus formation was observed throughout the healing process from radiographic assessment. Apparent mechanical property increased at 8 weeks, but tissue-level mechanical property did not increase significantly until 12 weeks. Three PCs were extracted from microstructural parameters and material distribution, which accounted for 87.592% of the total variation. The regression results showed a significant relationship between PCs and mechanical parameters (R>0.8, P<0.05). Results of this study show that microstructure and material distribution based on micro-CT imaging could efficiently predict bone strength and reflect the bone remodeling process during fracture healing, which provides a basis for exploring the fracture healing mechanism and may be used as an approach for fractured bone strength assessment.

Morphological and mechanical alterations in articular cartilage and subchondral bone during spontaneous hip osteoarthritis in guinea pigs.

Objectives: This study aimed to investigate the morphological and mechanical changes in articular cartilage and subchondral bone during spontaneous hip osteoarthritis in guinea pigs. Materials and methods: Hip joints of guinea pigs were investigated at 1, 3, 6, and 9 months of age (hereafter denoted as 1 M, 3 M, 6 M, and 9 M, respectively; n = 7 in each group). Morphological and mechanical alterations during spontaneous hip osteoarthritis in guinea pigs were investigated. The alterations included the micromechanical properties of articular cartilage (stiffness and creep deformation), microstructure of the subchondral bone (bone mineral density, bone volume fraction, trabecular thickness, trabecular number, and trabecular separation), micromorphology of the articular cartilage, and surface nanostructure (grain size and roughness) of the articular cartilage and subchondral bone. Results: Micromechanical properties of articular cartilage in 1 M showed the lowest stiffness and highest creep deformation with no significant differences in stiffness or creep deformation amongst 3 M, 6 M, and 9 M. Articular cartilage thickness decreased with age. The earliest degeneration of articular cartilage occurred at 6 months of age, characterised by surface unevenness and evident chondrocytes reduction in micromorphology, as well as increased grain size and decreased roughness in nanostructure. No degeneration at micro- or nanostructure of subchondral bone was observed before 9 months. Conclusion: Morphological degeneration of cartilage occurred before degeneration of mechanical properties. Meanwhile, degeneration of cartilage occurred before degeneration of subchondral bone during hip osteoarthritis. The current study provided novel insights into the structural and micromechanical interaction of hip osteoarthritis, which can serve as a theoretical basis for understanding the formation and progression of osteoarthritis.

Additively Manufactured Multilevel Voronoi-Lattice Scaffolds with Bonelike Mechanical Properties.

Irregular porous scaffold through Voronoi tessellation based on global modeling demonstrated randomness to a certain degree and susceptibility to producing large processing deviations. A modeling method for new types of scaffolds based on periodic arrays of Voronoi unit cell was proposed in this study. These porous scaffolds presented controllable local cells and satisfactory mechanical properties. The topological structure of the Voronoi unit cell was controlled using three independent cell design factors (Voronoi polyhedron volume V, face-centered scaled factor F1, and body-centered scaled factor F2), and multilevel Voronoi-lattice scaffolds were constructed on the basis of periodic arrays of the Voronoi unit cell. Compressive test and simulation were combined to quantify the mechanical properties of scaffolds. The regression equations were established using the response surface method (RSM) to determine relationships between Voronoi unit cell design factors and structural characteristic parameters and mechanical properties. The same trends were observed in stress-strain curves of the compressive test and simulation. The mechanical properties of scaffolds can be appropriately quantified via simulation. Regression equations based on RSM can properly predict the structural characteristic parameters and mechanical properties of the scaffold. Compared with V, F1 and F2 exerted a stronger influence on the structural characteristic parameters and mechanical properties of the scaffold. The modeling method of the multilevel Voronoi-lattice scaffold based on the Voronoi unit cell was proposed in this study to design the porous scaffold and meet the requirements of human bone morphology, mechanical properties, and actual manufacturing by adjusting factors V, F1, and F2. The proposed method can provide a feasible strategy for designing implants with suitable and similar morphologies and mechanical properties to cancellous bone.

The adaptive response of rat tibia to different levels of peak strain and durations of experiment.

The osteogenic response of bone to compressive load is affected by peak strain and duration of experiment, however, combined effect of peak load and duration of experiment on the rat tibia axial loading model remains unclear. In this study, rat tibia axial loading models with different levels of peak strains and durations of experiments were established (peak loads: 10 N, 20 N and 40 N, experimental duration: 2 weeks and 4 weeks). Microcomputer tomography scanning (Micro-CT), compression mechanical test and bone tissue alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP) staining were used to investigate the effects of cyclic loading with different levels of peak strains and durations of experiments on osteogenic response of tibia in adult female Sprague-Dawley (SD) rats. This experiment demonstrates that high-level strain can significantly stimulate the osteogenic response of cancellous and cortical bones in a short period of time. However, under mid-level strain, prolonging the experiment time can significantly improve the microsturcture and macromechanical properties of bone, increase the number of osteoblasts, and achieve the goal of improving bone quality. In this study, two loading schemes of mid-level-long-term and high-level-short-term were analyzed, and the mechanical response of the tibia under two different loading schemes was investigated. It provided a theoretical basis for further promoting the study of improving human bone quality and preventing aging-related bone diseases through mechanical stimulation.

Effects of whole-body vibration at different periods on lumbar vertebrae in female rats.

The current study aimed to investigate the effects of whole-body vibration (WBV) before and after ovariectomy on lumbar vertebrae, and to observe whether the positive effects of WBV before and after ovariectomy on lumbar vertebrae in rats could be maintained after vibration stopped. Three-month-old female rats were divided into four groups (n = 45/group): control (CON), ovariectomy (OVA), WBV before ovariectomy (WBV-BO), and WBV after ovariectomy (WBV-AO) groups. For 1-8 weeks, WBV-BO group was subjected to vertical WBV. At the 9th week, the rats in WBV-BO, WBV-AO, and OVA groups were ovariectomized. During 11-18 weeks, WBV-AO group was subjected to vibration. For 19-26 weeks, no intervention was done for rats. The lumbar vertebrae were examined by Micro-CT, compressive test, creep test, and microindentation test. At the 8th week, the displacement of the L1-L2 annulus fibrosus in WBV-BO group was 18% smaller compared with CON group (p<0.05). At the 18th week, the elastic modulus of the L5 vertebral body in WBV-BO and WBV-AO groups was 53% and 57% higher than that in CON group, respectively (p<0.05); the displacement of the L1-L2 annulus fibrosus in WBV-BO group was 25% smaller than those in the other groups (p<0.05). At the 26th week, there was no significant difference in the displacement of the L1-L2 annulus fibrosus between WBV-BO group and other groups (p>0.05); the elastic modulus of the L5 vertebral body had no significant difference between WBV-AO group and CON group (p>0.05). Our results demonstrated that WBV before ovariectomy effectively prevented disc degeneration with significant effects up to 8 weeks after ovariectomy. The vertebral mechanical properties could be significantly improved by WBV after ovariectomy, but the residual effect did not maintain after WBV stopped.

An adaptation model for trabecular bone at different mechanical levels.

BACKGROUND: Bone has the ability to adapt to mechanical usage or other biophysical stimuli in terms of its mass and architecture, indicating that a certain mechanism exists for monitoring mechanical usage and controlling the bone's adaptation behaviors. There are four zones describing different bone adaptation behaviors: the disuse, adaptation, overload, and pathologic overload zones. In different zones, the changes of bone mass, as calculated by the difference between the amount of bone formed and what is resorbed, should be different. METHODS: An adaptation model for the trabecular bone at different mechanical levels was presented in this study based on a number of experimental observations and numerical algorithms in the literature. In the proposed model, the amount of bone formation and the probability of bone remodeling activation were proposed in accordance with the mechanical levels. Seven numerical simulation cases under different mechanical conditions were analyzed as examples by incorporating the adaptation model presented in this paper with the finite element method. RESULTS: The proposed bone adaptation model describes the well-known bone adaptation behaviors in different zones. The bone mass and architecture of the bone tissue within the adaptation zone almost remained unchanged. Although the probability of osteoclastic activation is enhanced in the overload zone, the potential of osteoblasts to form bones compensate for the osteoclastic resorption, eventually strengthening the bones. In the disuse zone, the disuse-mode remodeling removes bone tissue in disuse zone. CONCLUSIONS: The study seeks to provide better understanding of the relationships between bone morphology and the mechanical, as well as biological environments. Furthermore, this paper provides a computational model and methodology for the numerical simulation of changes of bone structural morphology that are caused by changes of mechanical and biological environments.

Strain Distribution Evaluation of Rat Tibia under Axial Compressive Load by Combining Strain Gauge Measurement and Finite Element Analysis.

This study is aimed at providing an effective method for determining strain-load relationship and at quantifying the strain distribution within the whole tibia under axial compressive load on rats. Rat tibial models with axial compressive load were designed. Strains in three directions (0°, 45°, and 90°) at the proximal shaft of the tibia were measured by using a strain gauge rosette, which was used to calculate the maximum and minimum principal strains. Moreover, the strain at the midshaft of the tibia was measured by a single-element strain gauge. The slopes of the strain-load curves with different peak loads were calculated to assess the stability of the strain gauge measurement. Mechanical environment in the whole tibia by the axial compressive load was quantified using finite element analysis (FEA) based on microcomputed tomography images. The von Mises elastic strain distributions of the whole tibiae were evaluated. Slopes of the strain-load curves showed no significant differences among different peak loads (ANOVA; P > 0.05), indicating that the strain-load relationship obtained from the strain gauge measurement was reasonable and stable. The FEA results corresponded to the experimental results with an error smaller than 15% (paired Student's t-test, P > 0.05), signifying that the FEA can simulate the experiment reasonably. FEA results showed that the von Mises elastic strain was the lowest in the middle and gradually increased to both sides along the lateral direction, with the maximal von Mises elastic strain being observed on the posterior side under the distal tibiofibular synostosis. The method of strain gauge measurements and FEA used in this study can provide a feasible way to obtain the mechanical environment of the tibiae under axial compressive load on the rats and serve as a reference for further exploring the mechanical response of the bone by axial compressive load.

Whole body vibration with rest days could improve bone quality of distal femoral metaphysis by regulating trabecular arrangement.

Low-magnitude, high-frequency vibration (LMHFV) with rest days (particularly seven rest days) was considerably effective in improving the morphological and mechanical properties of rat proximal femur. However, current knowledge is limited regarding the possible benefit of this mechanical regimen to other bone sites and whether the optimal rest days are the same. This study followed our previous experiment on LMHFV loading with rest days for three-month-old male Wistar rats. The experiment involved seven groups, namely, vibrational loading for X day followed with X day rest (X=1, 3, 5, 7), daily vibrational loading, tail suspension and baseline control. Micro-computed tomography (micro-CT) scanning was used to evaluate the microarchitecture of the distal femoral trabecular bone. Micro-CT image-based microfinite element analysis was performed for each distal femoral metaphysis. LMHFV with rest days substantially changed the trabecular arrangement from remarkably plate-like to rod-like. Vibrational loading with 1 day rest was substantially effective in improving the architecture and apparent- and tissuelevel mechanical properties of the rat distal femoral metaphysis. This study may provide an improved understanding of the sitespecific responses of bone tissue to LMHFV with rest days for a substantially effective therapy of a targeted bone site.

Finite Element Investigation of the Effects of the Low-Frequency Vibration Generated by Vehicle Driving on the Human Lumbar Mechanical Properties.

Long-term exposure to low-frequency vibration generated by vehicle driving impairs human lumbar spine health. However, few studies have investigated how low-frequency vibration affects human lumbar mechanical properties. This study established a poroelastic finite element model of human lumbar spinal segments L2-L3 to perform time-dependent vibrational simulation analysis and investigated the effects of different vibrational frequencies generated by normal vehicle driving on the lumbar mechanical properties in one hour. Analysis results showed that vibrational load caused more injury to lumbar health than static load, and vibration at the resonant frequency generated the most serious injury. The axial effective stress and the radial displacement in the intervertebral disc, as well as the fluid loss in the nucleus pulposus, increased, whereas the pore pressure in the nucleus pulposus decreased with increased vibrational frequency under the same vibrational time, which may aggravate the injury degree of human lumbar spine. Therefore, long-term driving on a well-paved road also induces negative effects on human lumbar spine health. When driving on a nonpaved road or operating engineering machinery under poor navigating condition, the auto seat transmits relatively high vibrational frequency, which is highly detrimental to the lumbar spine health of a driver.

Morphological and Microstructural Alterations of the Articular Cartilage and Bones during Treadmill Exercises with Different Additional Weight-Bearing Levels.

The aim of this study was to investigate the morphological and microstructural alterations of the articular cartilage and bones during treadmill exercises with different exercise intensities. Sixty 5-week-old female rats were randomly divided into 10 groups: five additional weight-bearing groups (WBx) and five additional weight-bearing with treadmill exercise groups (EBx), which were subjected to additional weight bearing of x% (x = 0, 5, 12, 19, and 26) of the corresponding body weight of each rat for 15 min/day. After 8 weeks of experiment, the rats were humanely sacrificed and their bilateral intact knee joints were harvested. Morphological analysis of the cartilages and microcomputed tomography evaluation of bones were subsequently performed. Results showed that increased additional weight bearing may lead to cartilage damage. No significant difference was observed among the subchondral cortical thicknesses of the groups. The microstructure of subchondral trabecular bone of 12% and 19% additional weight-bearing groups was significantly improved; however, the WB26 and EB26 groups showed low bone mineral density and bone volume fraction as well as high structure model index. In conclusion, effects of treadmill exercise on joints may be associated with different additional weight-bearing levels, and exercise intensities during joint growth and maturation should be selected reasonably.

Effects of Scan Resolutions and Element Sizes on Bovine Vertebral Mechanical Parameters from Quantitative Computed Tomography-Based Finite Element Analysis.

Quantitative computed tomography-based finite element analysis (QCT/FEA) has been developed to predict vertebral strength. However, QCT/FEA models may be different with scan resolutions and element sizes. The aim of this study was to explore the effects of scan resolutions and element sizes on QCT/FEA outcomes. Nine bovine vertebral bodies were scanned using the clinical CT scanner and reconstructed from datasets with the two-slice thickness, that is, 0.6 mm (PA resolution) and 1 mm (PB resolution). There were significantly linear correlations between the predicted and measured principal strains (R2 > 0.7, P < 0.0001), and the predicted vertebral strength and stiffness were modestly correlated with the experimental values (R2 > 0.6, P < 0.05). Two different resolutions and six different element sizes were combined in pairs, and finite element (FE) models of bovine vertebral cancellous bones in the 12 cases were obtained. It showed that the mechanical parameters of FE models with the PB resolution were similar to those with the PA resolution. The computational accuracy of FE models with the element sizes of 0.41 × 0.41 × 0.6 mm3 and 0.41 × 0.41 × 1 mm3 was higher by comparing the apparent elastic modulus and yield strength. Therefore, scan resolution and element size should be chosen optimally to improve the accuracy of QCT/FEA.

Multi-Level Assessment of Fracture Calluses in Rats Subjected to Low-Magnitude High-Frequency Vibration with Different Rest Periods.

The aim of this study was to investigate the influences of low-magnitude high-frequency vibration (LMHFV) with different rest period regimes (vibrational loading per day [with or without the loading divided into bouts]; or vibrational loading for 7 day followed by 7 day rest [with or without the loading divided into bouts]) on bone healing at multi-levels. Transverse fractures of rat bilateral tibias were established using a Kirschner wire inserted for fixation. The animals were randomly assigned to five groups (n = 7 for each group): four for vibrational groups by LMHFV with different rest period regimes and one for fractured model without mechanical loading. The macromechanical properties of the fractured tibias and the nanomechanical properties of the calluses were investigated through three-point bending and nanoindentation tests, respectively. Atomic force microscopy (AFM) was performed to analyze the nanostructure of the calluses. Micro-computed tomography (micro-CT) scanning was conducted to evaluate the microarchitecture of the calluses. The serum concentration of osteocalcin (OG) and tartrate-resistant acid phosphatase 5b (TRAP5b) were measured to assess the bone formation and resorption rates, respectively. Significantly higher values of failure load and elastic modulus were observed in DL (vibrational loading for 15 min per day) and DLR (vibrational loading per day in which three bouts of 5 min of vibration were separated by 4 h) than FBC (fractured model without mechanical loading) at macro-level (P < 0.05). The results of nanoindentation test showed the highest values of indentation modulus and hardness in DLR (significantly higher than FBC; P < 0.05); besides, higher value of hardness was also observed in DL (significantly higher than FBC; P < 0.05). Though AFM imaging showed no significant differences in grain sizes between the vibrational groups and FBC, roughness of DLR showed the highest value, i.e. it was significantly higher than that in FBC (P < 0.05). For microarchitectural parameters obtained from micro-CT imaging, tissue mineral density (TMD) of DLR and VL7 (vibrational loading for 7 day followed by a 7-day rest, 15 min per day during vibrational periods) were significantly higher than that in FBC (P < 0.05), but no significant differences in other parameters were observed between vibrational groups and FBC. There were no significant differences of OG between vibrational groups and FBC; however, FBC showed significantly higher TRAP5b concentration than all vibrational groups (P < 0.05). The results show that LMHFV with different rest period regimes not only altered the macro- and nano-level bone mechanical properties but also influenced the TMD of calluses and nano-level spatial arrangement (roughness) significantly. The most significant effect of LMHFV with different rest period regimes was observed in DLR, which indicated that both osteogenic accumulation and cellular resensitization can be satisfied under this regimen. Hence, the DLR group demonstrated a great potential in clinical applications.

Age-related regional deterioration patterns and changes in nanoscale characterizations of trabeculae in the femoral head.

This study aimed to investigate the mechanical properties and features of bone materials at the nanoscale level in different regions of the femoral head in elderly patients with femoral neck fracture. Ten femoral heads from female patients with femoral neck fractures were extracted during surgery (five for the Aged group, aged 65-66 years; five for the Advanced aged group, aged 85-95 years). The femoral head was divided into three equal layers (anterior, central, and posterior) in the coronal view, and each layer was segmented into five regions (superior, central, inferior, medial, and lateral). Nanoindentation testing and atomic force microscopy imaging were used to study the mechanical properties and surface morphology of the specimens. No statistical differences in grain size were found between age groups, which suggested that the nanostructure of trabeculae in the femoral heads of postmenopausal women cannot be used to predict age-related bone loss and fracture risk. Mechanical properties in the longitudinal direction deteriorated more quickly than those in the transverse direction for the whole femoral head. Comparisons between layers showed a higher deterioration rate with aging in the anterior layer than in other layers. In different regions, mechanical properties of the medial and lateral regions deteriorated more quickly than those in the three other regions, and deterioration in the longitudinal direction was more serious than that in the transverse direction. The regional deterioration patterns and material properties with aging observed in this study contribute to an understanding of the age-related fracture mechanism and provide a basis for predicting age-related fracture risk and decreasing early fixation failure in the proximal femur.

Seven day insertion rest in whole body vibration improves multi-level bone quality in tail suspension rats.

OBJECTIVE: This study aimed to investigate the effects of low-magnitude, high-frequency vibration with rest days on bone quality at multiple levels. METHODS: Forty-nine three-month-old male Wistar rats were randomly divided into seven groups, namely, vibrational loading for X day followed by X day rest (VLXR, X = 1, 3, 5, 7), vibrational loading every day (VLNR), tail suspension (SPD), and baseline control (BCL). One week after tail suspension, rats were loaded by vibrational loading (35 Hz, 0.25 g, 15 min/day) except SPD and BCL. Fluorescence markers were used in all rats. Eight weeks later, femora were harvested to investigate macromechanical properties, and micro-computed tomography scanning and fluorescence test were used to evaluate microarchitecture and bone growth rate. Atomic force microscopy analyses and nanoindentation test were used to analyze the nanostructure and mechanical properties of bone material, respectively. Inductively coupled plasma optical emission spectroscopy was used for quantitative chemical analyses. RESULTS: Microarchitecture, mineral apposition rate and bone formation rate and macromechanical properties were improved in VL7R. Grain size and roughness were significantly different among all groups. No statistical difference was found for the mechanical properties of the bone material, and the chemical composition of all groups was almost similar. CONCLUSIONS: Low-magnitude, high-frequency vibration with rest days altered bone microarchitecture and macro-biomechanical properties, and VL7R was more efficacious in improving bone loss caused by mechanical disuse, which provided theoretical basis and explored the mechanisms of vibration for improving bone quality in clinics.