Characterization of the mechanical properties of bovine cortical bone treated with a novel tissue sterilization process

Over the past decade chemical processing and engineering of musculoskeletal tissue (tendon and bone) has improved dramatically. The use of bone allograft and xenograft in reconstructive orthopedic and maxillofacial surgeries is increasing, yet severe complications can occur if the material is contaminated in any way. A novel tissue sterilization process, BioCleanse^®, has been developed to clean and sterilize musculoskeletal tissue for implantation. The present study was designed to determine the effect of this novel cleaning process on the biomechanical properties of bovine cortical bone prior to implantation. The mechanical properties of treated bovine bone material were compared to human samples with respect to failure under compression, shear and three-point bending. The data demonstrate that bovine bone treated with the novel sterilization procedure has favorable biomechanical properties compared to that of human bone treated in a similar fashion.     

Biomechanical comparison of human bone-patellar tendon-bone grafts after sterilization with peracetic acid ethanol

Recent reports of disease transmission following ACL reconstruction with fresh-frozen non-sterilized allografts have highlighted the need for new sterilization techniques that do not impair the mechanical properties as it was shown for most of the current sterilization techniques. In this in-vitro biomechanical study, it was investigated if peracetic acid ethanol sterilization (PES) has any adverse effects on the mechanical properties of human bone-patellar tendon-bone grafts (BPTB). Paired human BPTB grafts either underwent PES or were used as fresh-frozen non-sterilized grafts. Viscoelastic properties (strain, creep) were analyzed during cyclic submaximal loading and mechanical properties were investigated during load-to-failure (LTF) testing. It was found that there were no differences in viscoelastic and mechanical properties between both groups. The findings of this study provide baseline data for future in vitro and in vivo analyses of this promising new sterilization technique for soft-tissue allografts.     

Time Related Changes of Mineral and Collagen and Their Roles in Cortical Bone Mechanics of Ovariectomized Rabbits

As cortical bone has a hierarchical structure, the macroscopic bone strength may be affected by the alterations of mineral crystal and collagen, which are main components of cortical bone. Limited studies focused on the time related alterations of these two components in osteoporosis, and their contributions to bone mechanics at tissue level and whole-bone level. Therefore, the purpose of this study was to elucidate the time related changes of mineral and collagen in cortical bone of ovariectomized (OVX) rabbits, and to relate these changes to cortical bone nanomechanics and macromechanics. 40 Rabbits (7-month-old) were randomly allocated into two groups (OVX and sham). OVX group received bilateral ovariectomy operation. Sham group received sham-OVX operation. Cortical bone quality of five rabbits in each group were assessed by DXA, μCT, nanoindentation, Fourier transform infrared (FTIR) spectroscopy and biomechanical tests (3-point bending of femoral midshaft) at pre-OVX, 4, 6, and 8 weeks after OVX. As time increased from pre-OVX to 8 weeks, the mineral to matrix ratio decreased with time, while both collagen crosslink ratio and crystallinity increased with time in OVX group. Elastic modulus and hardness measured by nanoindentation, whole-bone strength measured by biomechanical tests all decreased in OVX group with time. Bone material properties measured by FTIR correlated well with nano or whole-bone level mechanics. However, bone mineral density (BMD), structure, tissue-level and whole-bone mechanical properties did not change with age in sham group. Our study demonstrated that OVX could affect the tissue-level mechanics and bone strength of cortical bone. And this influence was attributed to the time related alterations of mineral and collagen properties, which may help us to design earlier interventions and more effective treatment strategies on osteoporosis.     

Quantification of growth factors in allogenic bone grafts extracted with three different methods

Background Bony allografts are used for defect filling. A reliable sterilization method is the peracetic acid–ethanol sterilization procedure (PES). Several studies showed the antimicrobiological efficacy of this method. Aim of this study was the quantification of growth factors necessary for bone formation in PES sterilized allografts (n = 9). Methods To extract the growth factors from the tissue three different methods were used: (a) use of collagenase 1 for extraction, (b) incubation of the material in a proteinase inhibitor cocktail (Complete), and (c) extraction with guanidine HCl. The supernatants from the different methods were analyzed for the total protein concentration and different growth factors. Results The extraction with guanidine HCl resulted in the highest amount of protein measurable in the supernatants of the samples. For comparison of the individual growth factor values the results were normalized to the protein content. The highest growth factor amount/protein was detectable for BMP-2 using the GndHCL method followed by FGFa, IGF-I, TGF-β1, VEGF, and PDGF. Comparing the three extraction methods, significant differences were measured for the individual growth factor content. Conclusion PES sterilized bony allografts contain several growth factors. Depending on the extraction method, the quantity of the analyzed growth factors varies.     

Development and validation of a subject-specific finite element model of a human clavicle

This study aimed to develop and validate a finite element (FE) model of a human clavicle which can predict the structural response and bone fractures under both axial compression and anterior–posterior three-point bending loads. Quasi-static non-injurious axial compression and three-point bending tests were first conducted on a male clavicle followed by a dynamic three-point bending test to fracture. Then, two types of FE models of the clavicle were developed using bone material properties which were set to vary with the computed tomography image density of the bone. A volumetric solid FE model comprised solely of hexahedral elements was first developed. A solid-shell FE model was then created which modelled the trabecular bone as hexahedral elements and the cortical bone as quadrilateral shell elements. Finally, simulations were carried out using these models to evaluate the influence of variations in cortical thickness, mesh density, bone material properties and modelling approach on the biomechanical responses of the clavicle, compared with experimental data. The FE results indicate that the inclusion of density-based bone material properties can provide a more accurate reproduction of the force–displacement response and bone fracture timing than a model with uniform bone material properties. Inclusion of a variable cortical thickness distribution also slightly improves the ability of the model to predict the experimental response. The methods developed in this study will be useful for creating subject-specific FE models to better understand the biomechanics and injury mechanism of the clavicle.     

Intra- and interindividual differences in tensile strength of human bone

In a biomechanical study we evaluated the stability of human femurs. In order to document the inter- and intraindividual differences in bending stability, we tested 200 bone samples, harvested from the femurs of 5 multiorgan donors. The bending tests were performed on a material testing machine (ZWICK) under standardized conditions. Our results revealed significant differences in the stability of the femurs of the 5 multiorgan donors. Furthermore, there was a significant difference in breaking strength at different levels of the same bone. Clinical relevance: In studies aimed at evaluating the primary bony stability of alloimplants after different preservation or sterilization procedures, the above-mentioned results should be taken into consideration.     

Mineral density and biomechanical properties of bone tissue from male Arctic foxes (Vulpes lagopus) exposed to organochlorine contaminants and emaciation

We investigated the impact from dietary OC (organochlorine) exposure and restricted feeding (emaciation) on bone mineral density (BMD; g hydroxy-apatite cm(-2)) in femoral, vertebrate, skull and baculum osteoid tissue from farmed Arctic blue foxes (Vulpes lagopus). For femur, also biomechanical properties during bending (displacement [mm], load [N], energy absorption [J] and stiffness [N/mm]) were measured. Sixteen foxes (EXP) were fed a wet food containing 7.7% OC-polluted minke whale (Balaenoptera acutorostrata) blubber in two periods of body fat deposition (Aug-Dec) and two periods of body fat mobilisation (Jan-July) in which the food contained less energy and only 2% blubber. SigmaOC food concentration in the food containing 7.7% whale blubber was 309 ng/g wet mass. This corresponded to a SigmaOC exposure of ca. 17 microg/kg body mass/d and a responding SigmaOC residue in subcutaneous adipose tissue of ca. 1700 ng/g live mass in the 8 EXP fat foxes euthanized after 16 months. A control group (CON) composed of 15 foxes were fed equal daily caloric amounts of clean pork (Sus scrofa) fat. After 16 months, 8 EXP and 7 CON foxes were euthanized (mean body mass=9.25 kg) while the remaining 8 EXP and 8 CON foxes were given restricted food rations for 6 months resulting in a body weight reduction (mean body mass=5.46 kg). The results showed that only BMD(skull) vs. BMD(vertebrae) were significantly correlated (R=0.68; p=0.03; n=10) probably due to a similar composition of trabecular and cortical osteoid tissue. No difference in any of the BMD measurements or femoral biomechanical properties was found between EXP and CON foxes although BMD baculum was 1.6-folds lower in the EXP group. However, lean summer foxes had significantly lower femoral biomechanical properties measured as displacement (mm), energy absorption (J) and time (s) biomechanical properties than fat winter foxes (all p<0.004). This indicates lower stiffness and softer bones from fasting which is in agreement with previous studies. Further, it should be kept in mind when studying bone tissues in Arctic mammals also in order to avoid confounding effects from body condition.     

泼尼松灌胃与肌内注射对大鼠骨密度、骨生物力学性能及骨代谢的影响

目的观察泼尼松灌胃与肌内注射两种不同给药方法对大鼠骨密度、骨生物力学及骨代谢的影响。方法将45只SPF级雄性SD大鼠随机分为3组（正常组15只、灌胃组15只、肌内注射组15只）,其中正常组大鼠作为阴性对照,予0.9%生理盐水灌胃2 m L/d;灌胃组大鼠给予泼尼松0.5 mg/（kg·d）灌胃;肌内注射组大鼠给予泼尼松0.5 mg/（kg·d）;12周后测定离体的大鼠椎体骨密度及血清β-CTX、PINP水平变化,采用三点弯曲试验测量股骨皮质骨最大载荷、弹性载荷、断裂载荷等生物力学指标。结果与正常组相比,灌胃组及肌内注射组大鼠椎骨骨密度值均显著性降低（P〈0.05）;与灌胃组相比,肌内注射组大鼠椎骨骨密度显著下降（P〈0.05）;与正常组相比,灌胃组及肌内注射组大鼠股骨的弹性载荷、最大载荷、断裂载荷均显著降低（P〈0.05）,肌内注射组与灌胃组大鼠的弹性载荷、最大载荷、断裂载荷相比差异无显著性（P〉0.05）。与正常组相比,灌胃组及肌内注射组大鼠中血清β-CTX水平均显著升高（P〈0.05）而PINP水平均显著降低（P〈0.05）,与灌胃组相比,肌内注射组大鼠血清β-CTX水平显著升高（P〈0.05）而PINP水平显著降低（P〈0.05）。骨组织切片HE染色显示：肌内注射组大鼠的骨小梁明显纤细疏松,造血组织明显减少,脂肪组织明显增多。结论泼尼松对大鼠的骨密度、骨生物力学及骨代谢指标都有影响,而肌内注射泼尼松比口服对骨密度、骨强度、骨代谢的影响更大,更易造成骨质疏松症。因此,建议临床使用泼尼松时选择口服作为给药方式更安全。     

Biomechanical properties of bones from rats treated with sevelamer

Sevelamer hydrochloride is used for ten years in patients on dialysis as a phosphate binder. We have previously shown that oral application of sevelamer prevents the bone loss and increases the bone volume in ovariectomized rats. In this study we further analysed the biomechanical properties of bones from rats treated with sevelamer utilizing a threepoint bending test to determine the mechanical properties of the cortical bone of the mid-shaft femur, while the indentation test was used to determine the mechanical properties of cancellous bone in the marrow cavity of the distal femoral metaphysis. Parameters analyzed included: maximum load (F(u)), stiffness (S), energy absorbed (W), toughness (T) and ultimate strength (sigma). The intrinsic properties, stress, elastic modulus and toughness were determined from measured maximum load, strains, stiffness, energy absorbed, outer and inner diameters, and calculated bone cross-sectional moment of inertia. Sevelamer was given to rats for 25 weeks with a content of 3% of sevelamer in a standard diet, starting immediately following ovariectomy (OVX). Animals were divided to the following groups: (1) Sham; (2) Sham + sevelamer 3%; (3) OVX; (4) OVX + sevelamer 3%. Our results showed that sevelamer particularly influenced the rat trabecular bone by increasing the maximum load for 26.2%, energy absorbed for 24.2% and the ultimate strength for 26.2% in sham animals treated with sevelamer 3%, as compared to sham rats. Sevelamer 3% in OVX rats also increased the maximum load for 71.4%, stiffness for 70.7%, energy absorbed for 55.9% and the ultimate strength for 71.3% as compared to OVX controls. In the three bending test sevelamer had a very little effect on preventing loss of bone strenght in the cortical bone. These results collectively suggest that sevelamer improves bone biomechanical properties, mainly affecting trabecular bone quality in both normal and ovariectomized rats.     

不同消毒方法对生物化PET 人工韧带的生物力学影响

目的:探讨环氧乙烷、高压蒸汽、60Co照射三种消毒方法,对生物化PET人工韧带的生物力学影响,为选取合适的消毒灭菌方法提供实验依据。方法:以表面改性的聚对苯二甲酸乙二醇酯纤维为材料,制备生物化PET人工韧带,分别给予环氧乙烷熏蒸、高压蒸汽、60Co照射消毒处理,并设未作消毒处理的空白对照组,分别进行生物力学测试,对测量结果行统计学比较分析。结果:高压蒸汽消毒后,韧带所能承受的最大拉力及第一次破裂力降低,且与对照组间的差异均有统计学意义(P<0.05);60Co照射消毒后,韧带所能承受的最大拉力降低,与对照组间的差异有统计学意义(P<0.05),但第一次破裂力的差异不明显(P>0.05);环氧乙烷消毒处理后,最大拉力、第一次破裂力与对照组之间无明显差异(P>0.05);各组间断裂伸长率无明显差异(P>0.05)。结论:高压蒸汽消毒后韧带样品的生物力学性能降低比较明显;采用60Co射线消毒方法对产品的力学性能影响较小,但进一步的消毒方法改进可能更为理想;环氧乙烷消毒法可作为生物化PET人工韧带的有效消毒方法。     

Mechanical strength of bone allografts subjected to chemical sterilization and other terminal processing methods

Infectious disease transmission through the use of human donor allografts can be a catastrophic complication in an otherwise straightforward surgical procedure. The use of bone allograft in reconstructive orthopedic surgeries is increasing, yet severe complications, including death, can result if the transplanted tissues transmit a communicable disease to the tissue recipient. The BioCleanse((R)) tissue sterilization process is a fully automated, low-temperature chemical sterilization process that renders allograft tissue sterile. The purpose of this study was to evaluate the effect of a chemical tissue sterilization process on the mechanical strength of cortical bone allografts prior to implantation. Cylindrical cortical bone specimens were harvested from seven human cadaver donors and treated either by: chemical sterilization alone; chemical sterilization and terminal sterilization by gamma irradiation; chemical sterilization, lyophilization, terminal sterilization by STERRAD and rehydration; or untreated. The specimens were tested to failure in axial compression, diametral compression, shear, or bending. There were no significant differences in ultimate stress, strain, or fracture energy between the chemically sterilized and control groups in any of the testing modes.     

Combining specimen-specific finite-element models and optimization in cortical-bone material characterization improves prediction accuracy in three-point bending tests

Although the beam theory is widely used for calculating material parameters in three-point bending test, it cannot accurately describe the biomechanical properties of specimens after the yield. Hence, we propose a finite element (FE) based optimization method to obtain accurate bone material parameters from three-point bending test. We tested 80 machined bovine cortical bone specimens at both longitudinal and transverse directions using three-point bending. We then adopted the beam theory and the FE-based optimization method combined with specimen-specific FE models to derive the material parameters of cortical bone. We compared data obtained using these two methods and further evaluated two groups of parameters with three-point bending simulations. Our data indicated that the FE models with material properties from the FE-based optimization method showed best agreements with experimental data for the entire force-displacement responses, including the post-yield region. Using the beam theory, the yield stresses derived from 0.0058% strain offset for the longitudinal specimen and 0.0052% strain offset for the transverse specimen are closer to those derived from the FE-based optimization method, compared to yield stresses calculated without strain offset. In brief, we conclude that the optimization FE method is more appropriate than the traditional beam theory in identifying the material parameters of cortical bone for improving prediction accuracy in three-point bending mode. Given that the beam theory remains as a popular method because of its efficiency, we further provided correction functions to adjust parameters calculated from the beam theory for accurate FE simulation.     

乙草胺的微生物降解及其对土壤磷脂脂肪酸特性的影响

通过监测不同培养条件下（灭菌、加选择抑菌剂、未灭菌）土壤中乙草胺残留数量和培养期间微生物标识物磷脂脂肪酸（PLFAs）含量的变化,探讨了以田间推荐量施用乙草胺时,不同微生物群落对乙草胺降解的相对贡献以及乙草胺对黑土微生物群落结构的影响.结果表明：乙草胺易于被微生物降解,其中,细菌对乙草胺的降解作用显著大于真菌,而真菌对乙草胺有更强的耐受能力.施入乙草胺后,各磷脂脂肪酸数量发生了显著变化,C14∶0、C16∶0和C18∶0所指示的土壤微生物量均有所下降;细菌PLFAs数量在培养初期显著降低,而后与对照的差异显著减小,表明细菌活性随着乙草胺的降解有所恢复;在施用乙草胺后的整个培养过程中,土壤真菌PLFAs数量始终低于对照,暗示着乙草胺对真菌的抑制可能是长期而不可逆的.     

Some mechanical properties of goose femoral cortical bone

The ultimate compressive strength and modulus of elasticity of femoral cortical bone from adult geese (Anser anser), were determined by sex and by quadrant by compressing small right circular cylinders which were 2.4 mm in height and 0.8 mm in diameter. The average ultimate compressive strength was 183 +/- 29 MPa. The average modulus of elasticity was 13.2 +/- 3.4 GPa. The bending strength and bending modulus of elasticity were determined by a three point bend test on rectangular prisms which had the approximate dimensions 0.75 mm X 0.75 mm X 25 mm. The average bending strength was 263 +/- 44 MPa while the average bending modulus was 19.6 +/- 3.1 GPa. The calcium content was determined by atomic absorption spectrophotometry and no correlation was found with the mechanical properties. The histology of the cortical bone was examined both quantitatively and qualitatively. A unique type of Haversian bone is described. Goose bone was found to be morphologically similar to adolescent human bone and to have mechanical properties similar to those of adult human bone.     

The Effects of Prolonged Cryopreservation on the Biomechanical Properties of Bone Allografts: A Microbiological, Histological and Mechanical Study

Bone allografting is the most common form of allotransplantation in modern medicine. Bone banking is usually the major part of most tissue banks throughout the world. Several years ago, many standards of bone banking were set empirically, and have never been evaluated. One particular parameter or standard was outdating graft materials after 5 years of storage. This study was conducted to evaluate the effect of prolonged cryopreservation on the biomechanical properties of bone allografts and establish whether graft materials become contaminated during long-term storage.Proximal humeral bone allografts were obtained from the bone bank after 1, 3 and 5 years of –80°C cryopreservation. Samples of each humeral head, i.e., cartilage, subchondral bone and spongy bone were histologically examined for inter- and intra-cellular changes. A three-point mechanical bending test was used on identical pieces of cortical bone to compare fresh and cryopreserved materials. Fresh-retrieved cortical bone using identically-sized segments, served as a control. Cultures were taken from each respective sample to determine contamination or sterility.Results of both the histological and mechanical testing showed that there were no significant, qualitative histological, or quantitative mechanical differences among the samples. All the cultures were negative. Therefore, based on this study's parameters, bone allografts can safely be used after a cryopreservation period of over 5 years and should not be discarded.     

Validation of the sterilization procedure of allogeneic avital bone transplants using peracetic acid-ethanol.

Different procedures are available to inactivate bacteria and fungi, including their spores, as well as viruses in human bone transplants. The most efficient methods are considered to be gamma irradiation and thermal inactivation as well as chemical sterilization methods like the peracetic acid-ethanol treatment (PES). Following national and international standards or draft standards, the antimicrobial effectiveness of this procedure was evaluated. Due to the standardizable size as well as the clinical relevance, defatted human spongiosa cuboids (15x15x15 mm) served as model system. After treatment with PES for 2 and 4 hours, respectively, the titre of living micro-organisms was determined in the supernatant and the cuboid. A reduction in the titre of viable micro-organisms below the detection level (reduction factor >5 log10) was already achieved after an incubation time of 2 hours (Staphylococcus aureus, Enterococcus faecium, Pseudomonas aeruginosa, Bacillus subtilis, Clostridium sporogenes, Mycobacterium terrae, Candida albicans as well as spores of Bacillus subtilis). No viable micro-organisms could be detected in any of the PES-treated test cuboids. Spores of Aspergillus niger were also completely inactivated. The PES procedure proved to be a reliable method for the sterilization of human bone transplants derived from spongiosa.     

Animal Modelling of Lumbar Corpectomy and Fusion and in vivo Growth of Spine Supporting Bone by Titanium Cage Implants: An Experimental Study

##正## In this study a lumbar spinal fusion animal model is established to assess the effect of spinal fusion cage,and explore theminimum area ratio of titanium cage section to vertebral section that ensures bone healing and biomechanical property.Lumbarcorpectomy was conducted by posterolateral approach with titanium cage implantation combined with plate fixation.Titaniumcages with the same length but different diameters were used.After implantation of titanium cages,the progress of bone healingwas observed and the bone biomechanical properties were measured,including deformation and displacement in axial compression,flexion,extension,and lateral bending motion.The factors affecting the in vivo growth of spine supporting body wereanalyzed.The results show that the area ratio of titanium cage section to vertebral section should reach 1/2 to ensure the bonehealing,sufficient bone intensity and biomechanical properties.Some bone healing indicators,such as BMP,suggest that there isa relationship between the peak time and the peak value of bone formation and metabolism markers and the bone healing strength.     

Effect of gamma sterilization on microhardness of the cortical bone tissue of bovine femur in presence of N-Acetyl-L-Cysteine free radical scavenger

Gamma sterilization is usually used to minimize the risk of infection transmission through bone allografts. However, it is believed that gamma irradiation affects the mechanical properties of allografts and free radical scavengers can be used to alleviate the radiation-induced degradation of these properties. The aim of this study was to investigate the radioprotective effects of N-Acetyl-L-Cysteine (NAC) free radical scavenger on the material properties of sterilized bovine cortical bone at microstructure level. Forty-two cortical tissue specimens were excised from three bovine femurs and irradiated to 35 and 70 kGy gamma rays in the presence of 5, 50, and 100 mM concentrations of NAC. The localized variations in microhardness were evaluated via indentation in the radial and longitudinal directions to examine different regions of the microstructures of the specimens, including the osteonal and interstitial tissues. A significant increase was observed in the hardness of osteonal, interstitial, and longitudinal combined microstructures exposed to 35 and 70 kGy radiations (P < 0.05), whereas a relative reduction of the hardness was observed in the radial direction. Furthermore, it was found that the application of 50 and 100 mM NAC during gamma irradiation significantly subsided the hardening in longitudinal combined microstructure. Moreover, the reduction of hardness in radial direction was suppressed in the presence of 100 mM of NAC. In conclusion, the results indicated that NAC free radical scavenger can protect the cortical bone against deteriorative effects of ionizing radiation and can be used to improve the material properties of sterilized allografts.     

The relationship between musculoskeletal stress markers and biomechanical properties of the humeral diaphysis

Musculoskeletal stress markers (MSM) at entheses and bone biomechanical properties are used in activity reconstructions. The effect of physical activity on bone biomechanical properties is well established but the relative role of physical activity on MSM is less well known. In this article, it is hypothesized that the same causal mechanisms should affect MSM development as those responsible for bone biomechanical properties. Further, there should be a correlation between MSMs and bone cross-sectional properties as both are considered to reflect physical activity. This was tested using three skeletal samples: early 20th century Finnish (Helsinki) and two medieval English (Blackgate and York) populations. Torsional/average bending rigidity (J) for four cross-sectional locations at 80, 65, 50, and 35% of humeral length from the distal end was calculated and pectoralis major, teres major, and deltoid were scored for MSM. Correlations between MSM and size-standardized J were significant for many comparisons, although they were stronger in males than in females, especially on the right side. In ANOVAs, sample was found to be a significant influence on the right side in both sexes. Using an aggregated MSM score, covariance between J and high MSM scores was again stronger in males. Covariance between J and MSM was found both at cross-sectional locations under muscle insertions and at more distant locations, demonstrating both direct and general effects of muscular loadings applied to diaphyses. Thus, the two types of skeletal markers appear to be related to similar underlying mechanical factors, but effects may also be sex- and sample-specific.     

Growth-dependent effects of dietary protein concentration and quality on the biomechanical properties of the diaphyseal rat femur

ObjectivesThis study compares the effects of feeding growing rats with increasing concentrations of casein (C) and wheat gluten (G), proteins that show different biological qualities, on the morphometrical and biomechanical properties of the femoral diaphysis.Materials and methodsFemale rats were fed with one of ten diets containing different concentrations (5–30%) of C and G between the 30th and 90th days of life (Control = C-20%). Biomechanical structural properties of the right femur middiaphysis were estimated using a 3-point bending mechanical test with calculation of some indicators of bone material properties.ResultsBody weight and length were affected by treatments, values being highest in rats fed the C-20% diet. G diets affected negatively both parameters. Changes in cross-sectional geometry (mid-diaphyseal cross-sectional and cortical areas, femoral volume, and rectangular moment of inertia) were positively related to the C content of the diet, while they were severely and negatively affected by G diets. Similar behaviors were observed in the bone structural properties (fracture load, yielding load, diaphyseal stiffness and elastic energy absorption). When values of strength and stiffness were normalized for body weight, the differences disappeared. The bone material quality indicators (elastic modulus, yielding stress, elastic energy absorption/volume) did not differ significantly among all studied groups. Femoral calcium concentration in ashes was not significantly different among groups.ConclusionThe clear differences in strength and stiffness of bone beams induced by dietary protein concentration and quality seemed to be the result of an induced subnormal gain in bone structural properties as a consequence of a correlative subnormal gain in bone growth and mass, yet not in bone material properties.