Shear strength and toughness of trabecular bone are more sensitive to density than damage

Microdamage occurs in trabecular bone under normal loading, which impairs the mechanical properties. Architectural degradation associated with osteoporosis increases damage susceptibility, resulting in a cumulative negative effect on the mechanical properties. Treatments for osteoporosis could be targeted toward increased bone mineral density, improved architecture, or repair and prevention of microdamage. Delineating the relative roles of damage and architectural degradation on trabecular bone strength will provide insight into the most beneficial targets. In this study, damage was induced in bovine trabecular bone samples by axial compression, and the effects on the mechanical properties in shear were assessed. The damaged shear modulus, shear yield stress, ultimate shear stress, and energy to failure all depended on induced damage and decreased as the architecture became more rod-like. The changes in ultimate shear strength and toughness were proportional to the decrease in shear modulus, consistent with an effective decrease in the cross-section of trabeculae based on cellular solid analysis. For typical ranges of bone volume fraction in human bone, the strength and toughness were much more sensitive to decreased volume fraction than to induced mechanical damage. While ultimately repairing or avoiding damage to the bone structure and increasing bone density both improve mechanical properties, increasing bone density is the more important contributor to bone strength.     

Biomechanical consequences of developmental changes in trabecular architecture and mineralization of the pig mandibular condyle

The purpose of the present study was to examine the changes in apparent mechanical properties of trabecular bone in the mandibular condyle during fetal development and to investigate the contributions of altering architecture, and degree and distribution of mineralization to this change. Three-dimensional, high-resolution micro-computed tomography (microCT) reconstructions were utilized to assess the altering architecture and mineralization during development. From the reconstructions, inhomogeneous finite element models were constructed, in which the tissue moduli were scaled to the local degree of mineralization of bone (DMB). In addition, homogeneous models were devised to study the separate influence of architectural and DMB changes on apparent mechanical properties. It was found that the bone structure became stiffer with age. Both the mechanical and structural anisotropies pointed to a rod-like structure that was predominantly oriented from anteroinferior to posterosuperior. Resistance against shear, also increasing with age, was highest in the sagittal plane. The reorganization of trabecular elements, which occurred without a change in bone volume fraction, contributed to the increase in apparent stiffness. The increase in DMB, however, contributed more dominantly. Incorporating the observed inhomogeneous distribution of mineralization decreased the apparent stiffness, but increased the mechanical anisotropy. This denotes that there might be a directional dependency of the DMB of trabecular elements, i.e. differently orientated trabecular elements might have different DMBs. In conclusion, the changes in DMB and its distribution are important to consider when studying mechanical properties during development and should be considered in other situations where differences in DMB are expected.     

Multiscale modeling of bone tissue with surface and permeability control

Natural biological materials usually present a hierarchical arrangement with various structural levels. The biomechanical behavior of the complex hierarchical structure of bone is investigated with models that address the various levels corresponding to different scales. Models that simulate the bone remodeling process concurrently at different scales are in development. We present a multiscale model for bone tissue adaptation that considers the two top levels, whole bone and trabecular architecture. The bone density distribution is calculated at the macroscale (whole bone) level, and the trabecular structure at the microscale level takes into account its mechanical properties as well as surface density and permeability. The bone remodeling process is thus formulated as a material distribution problem at both scales. At the local level, the biologically driven information of surface density and permeability characterizes the trabecular structure. The model is tested by a three-dimensional simulation of bone tissue adaptation for the human femur. The density distribution of the model shows good agreement with the actual bone density distribution. Permeability at the microstructural level assures interconnectivity of pores, which mimics the interconnectivity of trabecular bone essential for vascularization and transport of nutrients. The importance of this multiscale model relays on the flexibility to control the morphometric parameters that characterize the trabecular structure. Therefore, the presented model can be a valuable tool to define bone quality, to assist with diagnosis of osteoporosis, and to support the development of bone substitutes.     

Biomechanical properties across trabeculae from the proximal femur of normal and ovariectomised sheep

The elastic behaviour of trabecular bone is a function not only of bone volume and architecture, but also of tissue material properties. Variation in tissue modulus can have a substantial effect on the biomechanical properties of trabecular bone. However, the nature of tissue property variation within a single trabecula is poorly understood. This study uses nanoindentation to determine the mechanical properties of bone tissue in individual trabeculae. Using an ovariectomised ovine model, the modulus and hardness distribution across trabeculae were measured. In both normal and ovariectomised bone, the modulus and hardness were found to increase towards the core of the trabeculae. Across the width of the trabeculae, the modulus was significantly less in the ovariectomised bone than in the control bone. However, in contrast to this hardness was found not to differ significantly between the two groups. This study provides valuable information on the variation of mechanical material properties in healthy and diseased trabecular bone tissue. The results of the current study will be useful in finite element modelling where more accurate values of trabecular bone modulus will enable the prediction of the macroscale behaviour of trabecular bone.     

Magnetic resonance microscopy for the quantitative analysis of trabecular bone architecture

A major concern for long-term spaceflight is the effect of microgravity on bone structure and mass as a loss of cortical and trabecular bone volume and density, both of which can lead to decreased bone strength and an increased risk of bone fracture. Detailed analysis of the three-dimensional structure of trabecular bone, and its relation to bone strength has become feasible only recently using high-resolution 3D imaging techniques. In particular, magnetic resonance microscopy (MRM) has proved to be particularly useful for the ex vivo evaluation of the complex architecture of trabecular bone. In this study, we describe the use of two different MRM-based methods for the quantitative evaluation of the three-dimensional structure of trabecular bone explants and for the prediction of their biomechanical properties. The in vivo application of such methods is also discussed.     

Microstructure-based Finite Element Modelling and Characterisation of Bovine Trabecular Bone

1 Introduction The mechanical behaviour of trabecular bone is dependent on both the properties of individual trabeculae as well as the three-dimensional architecture into which they are arranged. Hence, a comprehensive study of trabecular bone requires the consideration of microscopic as well as macroscopic length scales. Nanoindentation is a technique particularly suited to the mechanical characterisation of single trabecula[1?3] owing to the impracticalities of isolating individual tra- becu…     

Direct mechanics assessment of elastic symmetries and properties of trabecular bone architecture

A method is presented to find orthotropic elastic symmetries and constants directly from the elastic coefficients in the overall stiffness matrix of trabecular bone test specimens. Contrary to earlier developed techniques, this method does not require pure orthotropic behavior or additional fabric measurements. The method uses high-resolution computer reconstructions of trabecular bone specimens as input for large-scale FE-analyses to determine all the 21 elastic coefficients in the overall stiffness matrix of the specimen, using a direct mechanics approach. An optimization procedure is then used to find the coordinate transformation that yields the best orthotropic representation of this matrix. The method is illustrated here relative to two trabecular bone specimens. The techniques developed here can be used to obtain a complete characterization of the mechanical properties of trabecular architecture. With the development of in vivo reconstruction techniques, even in vivo measurements will be possible.     

Interrelationship of trabecular mechanical and microstructural properties in sheep trabecular bone

The ability to evaluate fracture risk at an early time point is essential for improved prognostics as well as enhanced treatment in cases of bone loss such as from osteoporosis. Improving the diagnostic ability is inherent upon both high-resolution non-invasive imaging, and a thorough understanding of how the derived indices of structure and density relate to its true mechanical behavior. Using sheep femoral trabecular bone with a range of strength, the interrelationship of mechanical and microstructural parameters was analyzed using multi-directional mechanical testing and micro-computed tomography. Forty-five cubic trabecular bone samples were harvested from 23 adult female sheep, some of whom had received hind-limb vibratory stimuli over the course of 2 years with consequently enhanced mechanical properties. These samples were pooled into a low, medium, or high strength group for further analysis. The findings show that microCT indices that are structural in nature, e.g., structural model index (SMI) (r2=0.85, p<0.0001) is as good as more density oriented indices like bone volume/total volume (BV/TV) (r2=0.81, p<0.0001) in predicting the ultimate strength of a region of trabecular bone. Additionally, those indices more related to global changes in trabecular structure such as connectivity density (ConnD) or degree of anisotropy (DA) are less able to predict the mechanical properties of bone. Interrelationships of trabecular indices such as trabecular number (TbN), thickness (TbTh), and spacing (TbSp) provide clues as to how the trabecular bone will remodel to ultimately achieve differences in the apparent mechanical properties. For instance, the analysis showed that a loss of bone primarily affects the connectedness and overall number of trabeculae, while increased strength results in an increase of the overall thickness of trabeculae while not improving the connectedness. Certainly, the microCT indices studied are able to predict the bulk mechanical properties of a trabecular ROI well, leaving unaccounted only about 15-20% of its inherent variability. Diagnostically, this implies that future work on the early prediction of fracture risk should continue to explore the role of bone quality as the key factors or as an adjuvant to bone quantity (e.g., apparent density).     

How morphology predicts mechanical properties of trabecular structures depends on intra-specimen trabecular thickness variations

Two observations underlie this work. First, that the architecture of trabecular bone can accurately predict the mechanical stiffness characteristics of bone specimens when considering the combination of volume fraction and fabric, which is a measure of architectural anisotropy. Second, that the same morphological measures could not accurately predict the mechanical properties of porous structures in general. We hypothesize that this discrepancy can be explained by the special nature of trabecular bone as a structure in remodeling equilibrium relative to the external loads. We tested this hypothesis using a generic model of trabecular bone. Five series of 153 different architectures were created with this model. Each architecture was subjected to morphological analysis, and four different fabric measures were calculated to evaluate their effectiveness in characterizing the architecture. Relationships were determined relating morphology to the elastic constants. The quality of these relationships was tested by correlating the predicted elastic constants with those determined from finite element analysis. We found that the four fabric measures used could estimate the mechanical properties almost equally well. So the suggestion that fabric measures based on trabecular bone volume better represent the architecture than mean intercept length could not be affirmed. We conclude that for structures with equally sized elliptical voids the mechanical properties can be predicted well only if trabecular thickness variations within each structure are limited. These structures closely resemble previously developed models of trabecular bone. Furthermore, they are stiff in the principal fabric direction, hence, according to Cowin (J. Biomech. Eng. (108) (1986) 83), they are in remodeling equilibrium. These structures are also stiff over a large range of loading orientations, hence, are relatively insensitive to deviations in direction of loading.     

Comparison of HR-pQCT- and microCT-based finite element models for the estimation of the mechanical properties of the calcaneus trabecular bone

The calcaneus bone is formed of extensive trabecular bone and is therefore well suited to be used as an example of loaded bone to establish the ability of combining microfinite element (microFE) technique with high-resolution peripheral quantitative computed tomography (HR-pQCT) in determining its mechanical properties. HR-pQCT is increasingly used as a tool for in vivo bone clinical research, but its use has been limited to the distal radius and tibia. The goal of this study was to determine the applicability of HR-pQCT-derived microFE models of the calcaneus trabecular bone with 82 μm voxel size with reference to higher-resolution microCT-based models taken as gold standard. By comparing the outputs of microFE models generated from both HR-pQCT and microCT images of the trabecular bone of five calcaneus cadaveric specimens, it was found that the HR-pQCT-based models predicted mechanical properties for fracture load, total reaction force and von Mises stress are considerably different from microCT-based counterparts by 33, 64 and 70%, respectively. Also, the morphological analysis showed a comprehensive geometrical difference between HR-pQCT-based microFE models and their microCT-based equivalents. The results of the HR-pQCT-based models were found to have strong dependency on the threshold value chosen to binarise the images prior to finite element modelling. In addition, it was found that the voxel size has a strong impact on accuracy of imaged-based microFE models compared to other factors such as the presence of soft tissue and image scanning integration time. Therefore, although HR-pQCT has shown to be useful to predict overall structural and biomechanical changes, it is limited in providing local accurate biomechanical properties of trabecular bone and therefore should be used with caution when assessing bone remodelling through local changes of trabecular bone apposition and resorption in disease treatment monitoring.     

Does skeletal anatomy reflect adaptation to locomotor patterns? cortical and trabecular architecture in human and nonhuman anthropoids

Although the correspondence between habitual activity and diaphyseal cortical bone morphology has been demonstrated for the fore- and hind-limb long bones of primates, the relationship between trabecular bone architecture and locomotor behavior is less certain. If sub-articular trabecular and diaphyseal cortical bone morphology reflects locomotor patterns, this correspondence would be a valuable tool with which to interpret morphological variation in the skeletal and fossil record. To assess this relationship, high-resolution computed tomography images from both the humeral and femoral head and midshaft of 112 individuals from eight anthropoid genera (Alouatta, Homo, Macaca, Pan, Papio, Pongo, Trachypithecus, and Symphalangus) were analyzed. Within-bone (sub-articular trabeculae vs. mid-diaphysis), between-bone (forelimb vs. hind limb), and among-taxa relative distributions (femoral:humeral) were compared. Three conclusions are evident: (1) Correlations exists between humeral head sub-articular trabecular bone architecture and mid-humerus diaphyseal bone properties; this was not the case in the femur. (2) In contrast to comparisons of inter-limb diaphyseal bone robusticity, among all species femoral head trabecular bone architecture is significantly more substantial (i.e., higher values for mechanically relevant trabecular bone architectural features) than humeral head trabecular bone architecture. (3) Interspecific comparisons of femoral morphology relative to humeral morphology reveal an osteological "locomotor signal" indicative of differential use of the forelimb and hind limb within mid-diaphysis cortical bone geometry, but not within sub-articular trabecular bone architecture.     

Analysis of microstructural and mechanical alterations of trabecular bone in a simulated three-dimensional remodeling process

Bone remodeling is a complex dynamic process, which modulates both bone mass and bone microstructure. In addition to bone mass, bone microstructure is an important contributor to bone quality in osteoporosis and fragility fractures. However, the quantitative knowledge of evolution of three-dimensional (3D) trabecular microstructure in adaptation to the external forces is currently limited. In this study, a new 3D simulation method of remodeling of human trabecular bone was developed to quantitatively study the dynamic evolution of bone mass and trabecular microstructure in response to different external loading conditions. The morphological features of trabecular plate and rod, such as thickness and number density in different orientations were monitored during the remodeling process using a novel imaging analysis technique, namely Individual Trabecula Segmentation (ITS). We showed that the volume fraction and microstructures of trabecular bone including, trabecular type and orientation, were determined by the applied mechanical load. Particularly, the morphological parameters of trabecular plates were more sensitive to the applied load, indicating that they played the major role in the mechanical properties of the trabecular bone. Reducing the applied load caused severe microstructural deteriorations of trabecular bone, such as trabecular plate perforation, rod breakage, and a conversion from plates to rods.     

Sensitivity of damage predictions to tissue level yield properties and apparent loading conditions

High-resolution finite element models of trabecular bone failure could be used to augment current techniques for measuring damage in trabecular bone. However, the sensitivity of such models to the assumed tissue yield properties and apparent loading conditions is unknown. The goal of this study was to assess the sensitivity of the amount and mode (tension vs. compression) of tissue level yielding in trabecular bone to these factors. Linear elastic, high-resolution finite element models of nine bovine tibial trabecular bone specimens were used to calculate the fraction of the total tissue volume that exceeded each criterion for apparent level loading to the reported elastic limit in both on-axis and transverse compression and tension, and in shear. Four candidate yield criteria were studied, based on values suggested in the literature. Both the amount and the failure mode of yielded tissue were sensitive to the magnitudes of the tissue yield strains, the degree of tension-compression asymmetry of the yield criterion, and the applied apparent loads. The amount of yielded tissue was most sensitive to the orientation of the applied apparent loading, with the most tissue yielding for loading along the principal trabecular orientation and the least for loading perpendicular to it, regardless of the assumed tissue level yield criterion. Small changes in the magnitudes and the degree of asymmetry of the tissue yield criterion resulted in much larger changes in the amount of yielded tissue in the model. The results indicate that damage predictions based on high-resolution finite element models are highly sensitive to the assumed tissue yield properties. As such, good estimates of these values are needed before high-resolution finite element models can be applied to the study of trabecular bone damage. Regardless of the assumed tissue yield properties, the amount and type of damage that occurs in trabecular bone depends on the relative orientations of the applied apparent loads to the trabecular architecture, and this parameter should be controlled for both experimental and computational damage studies.     

Computational simulation of trabecular adaptation progress in human proximal femur during growth

There are a large number of clinical and experimental studies that analyzed trabecular architecture as a result of bone adaptation. However, only a limited amount of quantitative data is currently available on the progress of trabecular adaptation during growth. In this paper, we proposed a two-step numerical simulation method that predicts trabecular adaptation progress during growth using a recently developed topology optimization algorithm, design space optimization (DSO), under the hypothesis that the mechanisms of DSO are functionally equivalent to those of bone adaptation. We applied the proposed scheme to trabecular adaptation simulation in human proximal femur. For the simulation, the full trabecular architecture in human proximal femur was represented by a two-dimensional μFE model with 50 μm resolution. In Step 1, we determined a reference value that regulates trabecular adaptation in human proximal femur. In Step 2, we simulated trabecular adaptation in human proximal femur during growth with the reference value derived in Step 1. We analyzed the architectural and mechanical properties of trabecular patterns through iterations. From the comparison with experimental data in the literature, we showed that in the early growth stage trabecular adaptation was achieved mainly by increasing bone volume fraction (or trabecular thickness), while in the later stage of the development the trabecular architecture gained higher structural efficiency by increasing structural anisotropy with a relatively low level of bone volume fraction (or trabecular thickness). We demonstrated that the proposed numerical framework predicted the growing progress of trabecular bone that has a close correlation with experimental data.     

Predicting changes in mechanical properties of trabecular bone by adaptive remodeling

Because changes in the mechanical properties of bone are closely related to trabecular bone remodeling, methods that consider the temporal morphological changes induced by adaptive remodeling of trabecular bone are needed to estimate long-term fracture risk and bone quality in osteoporosis. We simulated bone remodeling using simplified and pig trabecular bone models and estimated the morphology of healthy and osteoporotic cases. We then displayed the fracture risk of the remodeled models based on a cumulative histogram from high stress. The histogram showed more elements had higher stresses in the osteoporosis model, indicating that the osteoporosis model had a greater risk.     

Effects of damage on the orthotropic material symmetry of bovine tibial trabecular bone

The macroscopic mechanical properties of trabecular bone can be predicted by its architecture using theoretical relationships between the elastic and architectural properties. Microdamage caused by overloading or fatigue decreases the apparent elastic moduli of trabecular bone requiring these relationships to be modified to predict the damaged elastic properties. In the case of isotropic damage, the apparent level elastic properties could be determined by multiplying all of the elastic constants by a single scalar factor. If the damage is anisotropic, the elastic constants may change by differing factors and the material coordinate system could become misaligned with the fabric coordinate system. High-resolution finite element models were used to simulate damage overloading on seven trabecular bone specimens subjected to pure shear strain in two planes. Comparison of the apparent elastic moduli of the specimens before and after damage showed that the reduction of the elastic moduli was anisotropic. This suggests that the microdamage within the specimens was inhomogeneous. However, after damage the specimens exhibited nearly orthotropic material symmetry as they did before damage. Changes in the orientation of the orthotropic material coordinate system were also small and occurred primarily in the transverse plane. Thus, while damage in trabecular bone is anisotropic, the material coordinate system remains aligned with the fabric tensor.     

Mechanisms of reduced implant stability in osteoporotic bone

The determining factors for the fixation of uncemented screws in bone are the bone-implant interface and the peri-implant bone. The goal of this work was to explore the role of the peri-implant bone architecture on the mechanics of the bone-implant system. In particular, the specific aims of the study were to investigate: (i) the impact of the different architectural parameters, (ii) the effects of disorder, and (iii) the deformations in the peri-implant region. A three-dimensional beam lattice model to describe trabecular bone was developed. Various microstructural features of the lattice were varied in a systematic way. Implant pull-out tests were simulated, and the stiffness and strength of the bone-implant system were computed. The results indicated that the strongest decrease in pull-out strength was obtained by trabecular thinning, whereas pull-out stiffness was mostly affected by trabecular removal. These findings could be explained by investigating the peri-implant deformation field. For small implant displacements, a large amount of trabeculae in the peri-implant region were involved in the load transfer from implant to bone. Therefore, trabecular removal in this region had a strong negative effect on pull-out stiffness. Conversely, at higher displacements, deformations mainly localized in the trabeculae in contact with the implant; hence, thinning those trabeculae produced the strongest decrease in the strength of the system. Although idealized, the current approach is helpful for a mechanical understanding of the role played by peri-implant bone.     

Incorporating tissue anisotropy and heterogeneity in finite element models of trabecular bone altered predicted local stress distributions

Trabecular bone is composed of organized mineralized collagen fibrils, which results in heterogeneous and anisotropic mechanical properties at the tissue level. Recently, biomechanical models computing stresses and strains in trabecular bone have indicated a significant effect of tissue heterogeneity on predicted stresses and strains. However, the effect of the tissue-level mechanical anisotropy on the trabecular bone biomechanical response is unknown. Here, a computational method was established to automatically impose physiologically relevant orientation inherent in trabecular bone tissue on a trabecular bone microscale finite element model. Spatially varying tissue-level anisotropic elastic properties were then applied according to the bone mineral density and the local tissue orientation. The model was used to test the hypothesis that anisotropy in both homogeneous and heterogeneous models alters the predicted distribution of stress invariants. Linear elastic finite element computations were performed on a 3 mm cube model isolated from a microcomputed tomography scan of human trabecular bone from the distal femur. Hydrostatic stress and von Mises equivalent stress were recorded at every element, and the distributions of these values were analyzed. Anisotropy reduced the range of hydrostatic stress in both tension and compression more strongly than the associated increase in von Mises equivalent stress. The effect of anisotropy was independent of the spatial redistribution high compressive stresses due to tissue elastic heterogeneity. Tissue anisotropy and heterogeneity are likely important mechanisms to protect bone from failure and should be included for stress analyses in trabecular bone.     

Multi-Scale Approach for the Evaluation of Bone Mineralization in Strontium Ranelate-Treated Diabetic Rats

Long-term diabetes mellitus can induce osteopenia and osteoporosis, an increase in the incidence of low-stress fractures, and/or delayed fracture healing. Strontium ranelate (SrR) is a dual-action anti-osteoporotic agent whose use in individuals with diabetic osteopathy has not been adequately evaluated. In this study, we studied the effects of an oral treatment with SrR and/or experimental diabetes on bone composition and biomechanics. Young male Wistar rats (half non-diabetic, half with streptozotocin/nicotinamide-induced diabetes) were either untreated or orally administered 625 mg/kg/day of SrR for 6 weeks. After sacrifice, femora from all animals were evaluated by a multi-scale approach (X-ray diffraction, Fourier transform infrared spectroscopy, inductively coupled plasma optical-emission spectrometry, static histomorphometry, pQCT, and mechanical testing) to determine chemical, crystalline, and biomechanical properties. Untreated diabetic animals (versus untreated non-diabetic) showed a decrease in femoral mineral carbonate content, in cortical thickness and BMC, in trabecular osteocyte density, in maximum load supported at rupture and at yield point, and in overall toughness at mid-shaft. Treatment of diabetic animals with SrR further affected several parameters of bone (some already impaired by diabetes): crystallinity index (indicating less mature apatite crystals); trabecular area, BMC, and vBMD; maximum load at yield point; and structural elastic rigidity. However, SrR was also able to prevent the diabetes-induced decreases in trabecular osteocyte density (completely) and in bone ultimate strength at rupture (partially). Our results indicate that SrR treatment can partially but significantly prevent some bone structural mechanical properties as previously affected by diabetes, but not others (which may even be worsened).     

Role of recombinant plasmid pEGFP-N1-IGF-1 transfection in alleviating osteoporosis in ovariectomized rats

Decreased levels of serum insulin-like growth factor-1 (IGF-1) have been proven to cause osteoporosis. Gene transfer of IGF-1 offers an attractive technology to treat skeletal metabolic disorders including osteoporosis, but the viral vectors are limited by their high antigenicity and immune response. Our purpose was to investigate the expression of a non-invasive vector, recombinant plasmid enhanced green fluorescent protein-N1 (pEGFP-N1) that transferred IGF-1 gene into ovariectomized (OVX) rats in vivo and evaluate the effect of this therapy on osteoporosis. OVX or sham operations were performed in 60 female, 7-month-old unmated SD rats. 12 weeks after OVX operation, the vectors were transfected to the 10-month-old rats and experimental data were detected from 48 h to 7 week after transfection. Our results showed that remarkable expression of fluorescence and serum IGF-1 was observed in the rats transfected by recombinant plasmids, indicating that IGF-1 gene was successfully transferred to OVX rats by injecting the vector through hydrodynamic method via the tail vein. The bone metabolism index including serum alkaline phosphatase, the histomorphometric parameters of lumbar vertebra including trabecular area percentage, trabecular thickness, trabecular number and trabecular separation, and the bone mineral density (BMD) and biomechanical parameters of lumbar vertebra including BMD, maximum condensing force, crushing strength in OVX rats transfected by pEGFP-N1-IGF-1 were improved remarkably compared with OVX+pEGFP-N1 rats, indicating that the transfection of recombinant plasmid pEGFP-N1-IGF-1 played a significant role in alleviating osteoporosis in rats induced by OVX. This encouraged a potential approach of IGF-1 gene therapy to the treatment of osteoporosis.