Nanoindentation analysis of the micromechanical anisotropy in mouse cortical bone

Casanova, Michele; Anna, Balmelli; Carnelli, Davide; Courty, Diana; Schneider, Philipp; Müller, Ralph

doi:10.1098/rsos.160971

Cited by 32 publications

(21 citation statements)

References 59 publications

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“…In this study a similar nanoindentation procedure was used as reported by Casanova et al (2017) for indentations on the mouse femur; while the indentation parameters were the same, the anatomical site, the age and the condition of the tissue were different. As expected, there was a difference of ∼26% between the E r found by Casanova et al (mouse femur, C57BL/6, female 22 weeks old, hydrated) and that found in this study (mouse tibia, C57BL/6, female 16 weeks old, dehydrated).…”

Section: Discussionmentioning

confidence: 99%

“…Indentations were performed with a Berkovich tip (Hysitron TI Primer nanoindenter, Bruker, USA) up to a maximum load of 6,000 µN that on our specimens lead to a penetration depth of ∼500 nm and radius of ∼3,500 nm (indentations within a lamella), as reported in a previous study where indentations were performed on the mouse femur (Casanova et al, 2017). The indentations were performed with the following parameters: loading time equal to 20.0 s (loading rate equal to 300 µN/s), holding time equal to 30.0 s, and unloading of 6.65 s (unloading rate equal to 902 µN/s).…”

Section: Nanoindentation Testsmentioning

confidence: 99%

“…For example, the nanoindentation properties of mouse bone tissue have been evaluated in the healing callus after fracture of the femur Morgan, 2008, 2009). Furthermore, nanoindentation tests have been performed in the femoral cortical bone of 22 weeks old female (Casanova et al, 2017) C57BL/6 mice or 16 weeks old (gender not reported) C57BL/6 (Pathak et al, 2011; with dynamics nanoindentation) and A/J (Pathak et al, 2012) mice (with spherical indenter).…”

Section: Introductionmentioning

confidence: 99%

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Regional Nanoindentation Properties in Different Locations on the Mouse Tibia From C57BL/6 and Balb/C Female Mice

Pepe

Oliviero

Cristofolini

et al. 2020

Front. Bioeng. Biotechnol.

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The local spatial heterogeneity of the material properties of the cortical and trabecular bone extracted from the mouse tibia is not well-known. Nevertheless, its characterization is fundamental to be able to study comprehensively the effect of interventions and to generate computational models to predict the bone strength preclinically. The goal of this study was to evaluate the nanoindentation properties of bone tissue extracted from two different mouse strains across the tibia length and in different sectors. Left tibiae were collected from four female mice, two C57BL/6, and two Balb/C mice. Nanoindentations with maximum 6 mN load were performed on different microstructures, regions along the axis of the tibiae, and sectors (379 in total). Reduced modulus (Er) and hardness (H) were computed for each indentation. Trabecular bone of Balb/C mice was 21% stiffer than that of C57BL/6 mice (20.8 ± 4.1 GPa vs. 16.5 ± 7.1 GPa). Moreover, the proximal regions of the bones were 13-36% less stiff than the mid-shaft and distal regions of the same bones. No significant differences were found for the different sectors for E r and H for Balb/C mice. The bone in the medial sector was found to be 8-14% harder and stiffer than the bone in the anterior or posterior sectors for C57BL/6 mice. In conclusion, this study showed that the nanoindentation properties of the mouse tibia are heterogeneous across the tibia length and the trabecular bone properties are different between Balb/C and C57BL/6 mice. These results will help the research community to identify regions where to characterize the mechanical properties of the bone during preclinical optimisation of treatments for skeletal diseases.

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Section: Discussionmentioning

confidence: 99%

Section: Nanoindentation Testsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Regional Nanoindentation Properties in Different Locations on the Mouse Tibia From C57BL/6 and Balb/C Female Mice

Pepe

Oliviero

Cristofolini

et al. 2020

Front. Bioeng. Biotechnol.

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“…However, to date, assessing the in situ longitudinal elastic modulus of single collagen fibril is still technical challenging. Moreover, Casanova et al (2017) also suggested that the transverse elastic modulus of the collagen network at the microscale level also has significant impact on the mechanical behavior of bone (R Soc Open Sci). In order to ensure comparability between the AFM nano-indentation tests and the tissue-level nano-indentation tests, we performed the transvers measurements at both the nanoscale (single collagen fibrils) and microscale level (tissue-level nano-indentation).…”

Section: Discussionmentioning

confidence: 99%

Disuse Impairs the Mechanical Competence of Bone by Regulating the Characterizations of Mineralized Collagen Fibrils in Cortical Bone

et al. 2019

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Bones are made of complex material comprising organic components and mineral hydroxyapatite, both of which formulate the unique hierarchical structure of bone and its mechanical properties. Bones are capable of optimizing their structure and mechanical properties according to the mechanical environment. Mineral loss is a well-known consequence of skeleton disuse. By contrast, the response of the non-mineral phase of bone, i.e., the collagen network, during disuse remain largely unknown. In this study, a tail-suspension mice model was used to induce bone loss. Atomic force microscopy-based imaging and indentation approaches were adopted to investigate the influence of disuse on the morphology and in situ mechanical behavior of the collagen fibrils, under both non-loaded and load-bearing conditions, in the cortical tibia of mice. The results indicate that disuse induced by hindlimb unloading did not alter the orientation and D-periodic spacing of the collagen fibril, but results in decreased collagen crosslinking which correlates with decreased elasticity and increased susceptibility to mechanical damage. More concretely, the collagen fibrils in the disused tibia were misaligned under mechanical loading. It therefore indicates that the disordered arrangement of the mineralized collagen fibrils is one of the characteristics of the weakened bone during elastic deformation. These findings reveals the unique adaptation regimes of the collagen fibrils in the cortical bone to disuse, as well as the deformation mechanisms of bone in the relevant pathological process at different scales.

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“…In separate studies, some researchers reported the mechanical properties of bone tissue in porcine, bovine, and mouse models, respectively. However, they did not comprehensively analyze the effects of age and region on mechanical properties and mechanical anisotropy [15, 16]. Canine bone was chosen because its tissue structure, systematic function, and fracture healing process were similar to humans and there was no published data on the mechanical properties of such bone to our knowledge [17, 18].…”

Section: Introductionmentioning

confidence: 99%

Analysis of Mechanical Properties and Mechanical Anisotropy in Canine Bone Tissues of Various Ages

Luo

Liao

Zhu

et al. 2019

BioMed Research International

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The effect of age on mechanical behavior and microstructure anisotropy of bone is often ignored by researchers engaged in the study of biomechanics. The objective of our study was to determine the variations in mechanical properties of canine femoral cortical bone with age and the mechanical anisotropy between the longitudinal and transverse directions. Twelve beagles divided into three age groups (6, 12, and 36 months) were sacrificed and all femurs were extracted. The longitudinal and transverse samples of cortical bone were harvested from three regions of diaphysis (proximal, central, and distal). A nanoindentation technique was used for simultaneously measuring force and displacement of a diamond tip pressed 2000nm into the hydrated bone tissue. An elastic modulus was calculated from the unloading curve with an assumed Poisson ratio of 0.3, while hardness was defined as the maximal force divided by the corresponding contact area. The mechanical properties of cortical bone were determined from 852 indents on two orthogonal cross-sectional surfaces. Mean elastic modulus ranged from 7.56±0.32 GPa up to 21.56±2.35 GPa, while mean hardness ranged from 0.28±0.057 GPa up to 0.84±0.072 GPa. Mechanical properties of canine femoral cortical bone tended to increase with age, but the magnitudes of these increase for each region might be different. The longitudinal mechanical properties were significantly higher than that of transverse direction (P<0.01). A significant anisotropy was found in the mechanical properties while there was no significant correlation between the two orthogonal directions in each age group (r2<0.3). Beyond that, the longitudinal mechanical properties of the distal region in each age group were lower than the proximal and central regions. Hence, mechanical properties in nanostructure of bone tissue must differ mainly among age, sample direction, anatomical sites, and individuals. These results may help a number of researchers develop more accurate constitutive micromechanics models of bone tissue in future studies.

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Nanoindentation analysis of the micromechanical anisotropy in mouse cortical bone

Cited by 32 publications

References 59 publications

Regional Nanoindentation Properties in Different Locations on the Mouse Tibia From C57BL/6 and Balb/C Female Mice

Regional Nanoindentation Properties in Different Locations on the Mouse Tibia From C57BL/6 and Balb/C Female Mice

Disuse Impairs the Mechanical Competence of Bone by Regulating the Characterizations of Mineralized Collagen Fibrils in Cortical Bone

Analysis of Mechanical Properties and Mechanical Anisotropy in Canine Bone Tissues of Various Ages

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