Effect of verapamil on bone mass, microstructure and mechanical properties in type 2 diabetes mellitus rats.

BACKGROUND: Verapamil was mainly used to treat hypertension, cardiovascular disease, inflammation and improve blood glucose in patients with diabetes, but its effects on bone mass, microstructure and mechanical properties were unclear. This study described the effects of verapamil on bone mass, microstructure, macro and nano mechanical properties in type 2 diabetic rats. METHODS: Rat models of type 2 diabetes were treated with verapamil at doses of 4, 12, 24 and 48 mg/kg/day by gavage respectively, twice a day. After 12 weeks, all rats were sacrificed under general anesthesia. Blood glucose, blood lipid, renal function and biochemical markers of bone metabolism were obtained by serum analysis, Micro-CT scanning was used to assess the microstructure parameters of cancellous bone of femoral head, three-point bending test was used to measure maximum load and elastic modulus of femoral shaft, and nano-indentation tests were used to measure indentation moduli and hardnesses of longitudinal cortical bone in femoral shaft, longitudinal and transverse cancellous bones in femoral head. RESULTS: Compared with T2DM group, transverse indentation moduli of cancellous bones in VER 24 group, longitudinal and transverse indentation moduli and hardnesses of cancellous bones in VER 48 group were significantly increased (p < 0.05). Furthermore, the effects of verapamil on blood glucoses, microstructures and mechanical properties in type 2 diabetic rats were dependent on drug dose. Starting from verapamil dose of 12 mg/kg/day, with dose increasing, the concentrations of P1NP, BMD, BV/TV, Tb. Th, Tb. N, maximum loads, elastic moduli, indentation moduli and hardnesses of femurs in rats in treatment group increased gradually, the concentrations of CTX-1 decreased gradually, but these parameters did not return to the level of the corresponding parameters of normal rats. Verapamil (48 mg/kg/day) had the best therapeutic effect. CONCLUSION: Verapamil treatment (24, 48 mg/kg/day) significantly affected nano mechanical properties of the femurs, and tended to improve bone microstructures and macro mechanical properties of the femurs, which provided guidance for the selection of verapamil dose in the treatment of type 2 diabetic patients.

Biochemical and Morphological Abnormalities of Subchondral Bone and Their Association with Cartilage Degeneration in Spontaneous Osteoarthritis.

This study aims to investigate how biochemical composition in subchondral bone (SB) relates to the sulfated glycosaminoglycan (sGAG) content of articular cartilage (AC) in the knee joint of guinea pigs from the early to moderate osteoarthritis (OA). Male Dunkin Hartley strain guinea pigs were grouped according to age (1, 3, 6, and 9 months, with 10 guinea pigs in each group). The biochemical properties of the AC and SB in the tibial plateau of the guinea pigs were determined through histology and Raman spectroscopy, respectively. Furthermore, the microstructures of the SB were investigated using micro-computed tomography (micro-CT) and histology. Increased thickness and bone mineral density (BMD) and decreased porosity were observed in the subchondral plate (SP) with the progression of spontaneous OA, accompanied by a decreasing trend in sGAG integrated optical density (IOD) of AC. Compared with the changes in the microstructure of subchondral bone, the content of sGAG was more correlated to the changes in the mineral/matrix ratio of subchondral bone. The mineralization of the matrix was significantly correlated to the content of sGAG compared with crystallinity/maturity and Type B carbonate substitution. PO43- ν1/Amide III was more correlated to the content of sGAG than PO43- ν1/Amide I, PO43- ν1/CH2 wag during the progression of spontaneous osteoarthritis. This study demonstrated that the mineralization of subchondral bone plays a crucial role in the pathogenesis of OA. Future studies may access to the mineralization of subchondral bone in addition to its microstructure in the study for pathogenesis and early diagnosis of osteoarthritis.

Study on the biomechanical responses of the loaded bone in macroscale and mesoscale by multiscale poroelastic FE analysis.

BACKGROUND: Bone is a hierarchically structured composite material, and different hierarchical levels exhibit diverse material properties and functions. The stress and strain distribution and fluid flow in bone play an important role in the realization of mechanotransduction and bone remodeling. METHODS: To investigate the mechanotransduction and fluid behaviors in loaded bone, a multiscale method was developed. Based on poroelastic theory, we established the theoretical and FE model of a segment bone to provide basis for researching more complex bone model. The COMSOL Multiphysics software was used to establish different scales of bone models, and the properties of mechanical and fluid behaviors in each scale were investigated. RESULTS: FE results correlated very well with analytical in macroscopic scale, and the results for the mesoscopic models were about less than 2% different compared to that in the macro-mesoscale models, verifying the correctness of the modeling. In macro-mesoscale, results demonstrated that variations in fluid pressure (FP), fluid velocity (FV), von Mises stress (VMS), and maximum principal strain (MPS) in the position of endosteum, periosteum, osteon, and interstitial bone and these variations can be considerable (up to 10, 8, 4 and 3.5 times difference in maximum FP, FV, VMS, and MPS between the highest and the lowest regions, respectively). With the changing of Young's modulus (E) in each osteon lamella, the strain and stress concentration occurred in different positions and given rise to microscale spatial variations in the fluid pressure field. The heterogeneous distribution of lacunar-canalicular permeability (klcp) in each osteon lamella had various influence on the FP and FV, but had little effect on VMS and MPS. CONCLUSION: Based on the idealized model presented in this article, the presence of endosteum and periosteum has an important influence on the fluid flow in bone. With the hypothetical parameter values in osteon lamellae, the bone material parameters have effect on the propagation of stress and fluid flow in bone. The model can also incorporate alternative material parameters obtained from different individuals. The suggested method is expected to provide dependable biological information for better understanding the bone mechanotransduction and signal transduction.

[Effect of artery pulse on the osteonal interstitial fluid flow behavior].

There are two main types of fluid in bone tissue: blood and interstitial fluid. The metabolism of cells mainly relies on the microenvironment of the interstitial fluid. Researches of osteonal fluid seepage behavior based on the microstructure of bone tissue have become a hot point. The aim of the present research work is to assess the effect of blood pressure oscillation on the osteonal interstitial fluid seepage behavior. We established finite element osteon models for a hollow and that considering blood pressure oscillation, respectively, with COMSOL Multiphysics software in order to compare their fluid flow behavior under the axial loading. The results predicted that the interstitial fluid pressure field was enlarged considering the blood pressure oscillation, while the velocity filed changed little. Specifically, the increase of blood pressure oscillatory amplitude could result in the increase of osteonal interstitial fluid pressure, while the blood pressure oscillatory frequency had limited effects on the osteonal pore fluid pressure. Moreover, the blood pressure oscillatory amplitude and frequency had no effect on the osteonal interstitial fluid velocity. The finite element model can be used for the study of the poroelastic behaviors of the osteon under non-axisymmetric loads and microcracks, and can also be a new way to study the mechanism of bone mechanotransduction and electromechanotransduction.

Biomechanical effects of typical lower limb movements of Chen-style Tai Chi on knee joint.

The load and stress distribution on cartilage and meniscus of the knee joint in typical lower limb movements of Chen-style Tai Chi (TC) and deep squat (DS) were analyzed using finite element (FE) analysis. The loadings for this analysis consisted of muscle forces and ground reaction force (GRF), which were calculated through the inverse dynamic approach based on kinematics and force plate measurements obtained from motion capture experiments. Thirteen experienced practitioners performed four typical TC movements, namely, single whip (SW), brush knee and twist step (BKTS), stretch down (SD), and part the wild horse's mane (PWHM), which exhibit lower posture and greater lower limb force compared to other TC styles. The results indicated that TC required greater lower limb muscle strength than DS, resulting in greater knee joint forces. The stress on the medial cartilage in SW and BKTS fell within a range conductive to maintaining the balance between anabolism and catabolism of cartilage matrix. This was due to the fact that SW and BKTS reduce the medial to total tibiofemoral contact force ratios through knee abduction, which may effectively alleviate mild medial knee osteoarthritis (KOA). However, the greater medial contact force ratios observed in SD and PWHM resulted in great contact stresses that may aggravate the pain of patients with KOA. To mitigate these effects, practitioners should consider elevating their postures appropriately to reduce knee flexion angles, especially during the single-leg support phase. This adjustment can decrease the required muscle strength, load and stress on the knee joint.

A Comparative Study on the Multiscale Mechanical Responses of Human Femoral Neck Between the Young and the Elderly Using Finite Element Method.

Background: Femoral neck fracture (FNF) is the most serious bone disease in the elderly population. The multiscale mechanical response is a key to predicting the strength of the femoral neck, assessing the risk of FNF, and exploring the role of mechanosensation and mechanotransmission in bone remodeling, especially in the context of aging bone. Methods: Multiscale finite element (FE) models of the proximal femur for both young and elderly people were developed. The models included organ scale (proximal femur), tissue scale (cortical bone), tissue element scale (osteon), and cell scale [osteocyte lacuna-canalicular network (LCN) and extracellular matrix (ECM), OLCEM]. The mechanical responses of cortical bone and osteocytes in the mid-femoral neck and the differences in mechanical responses between these two scales were investigated. Results: The mechanical responses of cortical bone and osteocyte showed significant differences between the elderly and the young. The minimum principal strains and mean SEDs of cortical bone in the elderly were 2.067-4.708 times and 3.093-14.385 times of the values in the young, respectively; the minimum principal strains and mean SEDs of osteocyte in the elderly were 1.497-3.246 times and 3.044-12 times of the values in the young, respectively; the amplification factors of minimum principal strain in the inferior (Inf), anterior (Ant), and posterior (Post) quadrants in the young were 1.241-1.804 times of the values in the elderly, but the amplification factor of minimum principal strain in the superior (Sup) quadrant was 87.4% of the value in the elderly; the amplification factors of mean SED in the young were 1.124-9.637 times of the values in the elderly. Conclusion: The mass and bone mineral density (BMD) of cortical bone in the femoral neck is closely related to the mechanical response of osteocytes, which provides a new idea for improving cortical bone quality. Perhaps cortical bone quality could be improved by stimulating osteocytes. Quadrantal differences of bone quality in the mid-femoral neck should be considered to improve fracture risk prediction in the future.

Prediction of Femoral Strength Based on Bone Density and Biochemical Markers in Elderly Men With Type 2 Diabetes Mellitus.

Purpose: Effects of bone density, bone turnover and advanced glycation end products (AGEs) on femoral strength (FS) are still unclear in patients with type 2 diabetes mellitus (T2DM). This study aims to assess and predict femoral strength and its influencing factors in elderly men with T2DM. Methods: T2DM patients (n = 10, mean age, 66.98 years) and age-matched controls (n = 8, mean age, 60.38 years) were recruited. Femoral bone mineral density (BMD) and serum biochemical indices of all subjects were measured. FS was evaluated through finite element analysis based on quantitative computed tomography. Multiple linear regression was performed to obtain the best predictive models of FS and to analyze the ability of predictors of FS in both groups. Results: FS (p = 0.034), HbA1c (p = 0.000) and fasting blood glucose (p = 0.000) levels of T2DM group were significantly higher than those of control group; however, the P1NP level (p = 0.034) was significantly lower. FS was positively correlated with femoral neck T score (FNTS) (r = 0.794, p < 0.01; r = 0.881, p < 0.01) in both groups. FS was correlated with age (r = -0.750, p < 0.05) and pentosidine (r = -0.673, p < 0.05) in T2DM group. According to multiple linear regression, FNTS and P1NP both contributed to FS in two groups. P1NP significantly improved the prediction of FS in both groups, but significant effect of FNTS on predicting FS was only presented in control group. Furthermore, pentosidine, age and HbA1c all played significant roles in predicting FS of T2DM. Conclusion: Femoral strength was higher in elderly men with T2DM, which might be caused by higher BMD and lower bone turnover rate. Moreover, besides BMD and bone formation level, AGEs, blood glucose and age might significantly impact the prediction of femoral strength in T2DM.

Multiscale mechanical responses of young and elderly human femurs: A finite element investigation.

BACKGROUND: Bone remodeling in the elderly is no longer balanced. As a result, the morphologies and mechanical properties of bone at different scales will change. These changes would affect the mechanical responses of bone, which might exacerbate the imbalance of bone remodeling and even cause age-related bone diseases. METHODS: Considering those changes, multiscale finite element (FE) models of bone in the young and the elderly were developed that included macroscale (proximal femur), mesoscale (cortical bone), microscale (Haversian system) and sub-microscale (osteocyte-lacuna-canaliculus-extracellular matrix system, OLCES). The stress and strain distributions at different scales and transmissions among different scales were investigated. RESULTS: The stresses of the elderly at macroscale, mesoscale and microscale were higher than those in the young by 23.7%, 62.5% and 8.0%, respectively, and the stresses of the elderly and the young at sub-microscale were almost the same. The strain of the elderly at macroscale, mesoscale, microscale and sub-microscale were higher than those in the young by 48.6%, 56.8%, 11.9% and 25.1%, respectively. The stress and strain transmission rates (ησand ηε) from mesoscale to microscale were decreased by 1.8%, and 2.5% than those from macroscale to mesoscale in the elderly, respectively; but increased by 13.8%, and 4.7% in the young, respectively. ηε from microscale to sub-microscale in the elderly was higher than that in the young by 21.3%. CONCLUSIONS: Degeneration of cortical bone mechanical property in the elderly causes increases in stress and strain at macroscale and mesoscale. The reduction of lacunar number in the elderly is not conducive to the mechanical transmission from mesoscale to microscale. The differences in stress and strain at microscale between the young and the elderly are smaller than those at macroscale or mesoscale. The strain stimulus sensed by osteocyte in the elderly is not weakened compared with that in the young.

Influence of non-enzymatic glycation on the mechanical properties of cortical bone.

Poor bone quality induced by non-enzymatic glycation (NEG) of bone tissue in patients with type 2 diabetes mellitus (T2DM) is regarded as the major factor of bone fragility and affecting bone mechanical properties. A comprehensive and systemic mechanical investigation for evaluating the effect of NEG on bone was still lacking. In order to provide additional information for the bone quality of T2DM, the effects of NEG on mechanical properties of cortical bone were investigated in terms of elastoplasticity, fracture toughness and viscoelasticity. All samples of cortical bone, including the samples of strength test (n = 20), fracture toughness test (n = 40, quasi-static and fall-like conditions with displacement rates of 10-3 mm/s and 10 mm/s, respectively) and stress relaxation test (n = 20), were harvested from bovine tibiae. The samples of each test were equally divided into incubated-control group and ribose-incubated group. All mechanical tests were performed after incubating all samples for 15 days. Post-yield strain (p = 0.014), post-yield energy (p < 0.0001) and damage fraction (p = 0.040) of ribose-incubated group were significantly lower than those of incubated-control group, but secant modulus (p = 0.029) of ribose-incubated group was significantly higher than that of incubated-control group. In quasi-static condition, the plastic contribution Jpl of fracture toughness (p = 0.043) of ribose-incubated group was significantly lower than that of incubated-control group. In fall-like condition, there were no differences in Jpl, elastic contribution Jel and J-integral in both two groups. The quasi-static Jel (p < 0.0001, p < 0.0001) of incubated-control and ribose-incubated groups and J-integral (p = 0.007) of incubated-control group were all significantly higher than those of fall-like condition. In stress relaxation test, initial modulus E0 (p = 0.040) and equilibrium modulus (p = 0.029) of ribose-incubated group were significantly higher than those of incubated-control group. Reductions of relaxation modulus, which were the differences between two adjacent time points within 700 s-3000 s for ribose-incubated group, were significantly lower than those of incubated-control group. NEG could decrease the post-yield properties and quasi-static facture toughness of cortical bone, especially the plastic contribution of quasi-static fracture toughness. It could also decrease the viscoelasticity of cortical bone. The present study confirmed the negative effects of NEG on the mechanical properties of cortical bone in terms of elastoplasticity, fracture toughness and viscoelasticity, but NEG had no significant effect on the fracture toughness of cortical bone at fall-like loading. These results provided more evidence for increased fragility of cortical bone in patients with T2DM.

Continuous subcutaneous insulin infusion ameliorates bone structures and mechanical properties in type 2 diabetic rats by regulating bone remodeling.

Continuous subcutaneous insulin infusion (CSII) is an intensive insulin therapy for patients with type 2 diabetes mellitus (T2DM) who have poor glycemic control, but its effect on T2DM-related bone disorder is unclear. This study described the possible mechanisms by which CSII affects bone remodeling, structures, and mechanical properties in T2DM rats. Herein, male rats (6-week-old) were assigned randomly to 4-week and 8-week administration groups, each of which included healthy control, T2DM, CSII, and Placebo groups. Then, metabolic markers, bone formation and resorption markers in serum and protein expressions of osteoclastogenesis regulators in tibias were detected. Meanwhile, microstructures, nanostructures, macro-mechanical properties, nano-mechanical properties, and mineral compositions in femurs were evaluated. 4-week later, CSII treatment restored circulatory metabolites, bone formation and resorption markers, and osteoclastogenesis regulators, improved certain bone microstructures, decreased matrix mineralization, and increased fracture toughness in T2DM rats. For 8-week group, CSII treatment restored bone formation and resorption markers, osteoclastogenesis regulators, and bone microstructures, besides improved bone mineral compositions and nanostructures, enhanced bone mechanical properties such as fracture toughness, maximum load, elastic modulus, indentation modulus and hardness. Collectively, 8-week CSII treatment is more conducive to ameliorating bone structures and mechanical properties in T2DM rats by regulating bone remodeling compared with 4-week CSII treatment, thus improving whole bone quality and providing valuable information for clinical prevention and treatment of T2DM-related bone disorders.

Effects of the microcrack shape, size and direction on the poroelastic behaviors of a single osteon: a finite element study.

PURPOSE: In this work, a finite element study is proposed by using the Comsol Multiphysics software to evaluate the effects of microcrack shape, size and direction on the poroelastic behaviors of a single osteon. METHODS: This finite element model is established by using the Comsol Multiphysics software, and we just focus on the comparison of the influences of those microcrack geometric parameters on the osteonal fluid pressure and velocity. RESULTS: The results show that: (1) microcracks in the osteon wall can induce a release of the fluid pressure, but enlarge the velocity in this region; (2) equal-area microcrack with ellipsoid-like shape produced a larger fluid pressure and velocity fields in the osteon than that of rectangular shape; (3) in the elliptic microcracks, the longer of the length (major semi-axis) induces a smaller fluid pressure and velocity amplitudes, whereas the width (minor axis) has little effect; (4) the direction of the microcracks (major axial direction) has an limited influence area around about 1/15 of the osteon cross-sectional area. CONCLUSIONS: This model permits the linking of the external loads and microcracks to the osteonal fluid pressure and velocity, which can be used for other purpose associate microcracks with the mechanotransduction and bone remodeling.