Jelena Potocnik

Age-related deterioration in trabecular bone mechanical properties at material level: Nanoindentation study of the femoral neck in women by using AFM

Despite general belief that the mechanical properties of bone material contribute to whole bone strength, it is still obscure what the age effects are on mechanical behavior of the bone material, particularly in the case of the femoral neck trabeculae. In this study, atomic force microscopy was used for imaging and measuring the size of mineral grains, as well as nano-scale mechanical characterization (nanoindentation) of the bone mineralized matrix of trabeculae, with the aim to explore the age effects on bone elasticity and give new insight into age-related bone fragility. The bone samples in this study comprised trabecular bone specimens of the femoral neck region, collected from eight skeletal healthy women (five young adults: 27-38yrs., three elderly: 83-94yrs.) at autopsy. Bone trabeculae in the elderly displayed a higher modulus and nanohardness, signifying a decreased amount of energy that can be accommodated by the bone tissue during loading. Regression analysis revealed that nearly 65% of variability in the bone matrix elastic modulus can be statistically explained by the changes in size of the matrix mineral grains. This study revealed that the bone trabeculae of elderly women express less elastic behavior at the material level, which makes them more vulnerable to unusual impact loads originating from a fall. The observed age-related structural and mechanical alteration at the bone material level adds new evidence for understanding why hip fractures are more frequent in elderly women.

Nanostructure and mineral composition of trabecular bone in the lateral femoral neck: implications for bone fragility in elderly women.

Despite interest in investigating age-related hip fractures, the determinants of decreased bone strength in advanced age are not clear enough. Hitherto it has been obscure how the aging process affects the femoral neck nanostructure and composition, particularly in the lateral subregion of the femoral neck, which is considered as a fracture-initiating site. The femoral bone samples used in this study were obtained at autopsy in 10 women without skeletal disease (five younger: aged 20-40 years, and five elderly: aged 73-94 years). Atomic force microscopy (AFM) was applied to explore the mineral grain size in situ in young vs. old trabecular bone samples from the lateral femoral neck. The chemical compositions of the samples were determined using inductively coupled plasma optical emission spectroscopy and direct current argon arc plasma optical emission spectrometry. Our AFM study revealed differences in trabecular bone nanostructure between young and elderly women. The mineral grain size in the trabeculae of the old women was larger than that in the young (median: 95 vs. 59nm), with a particular bimodal distribution: 45% were small grains (similar to the young) and the rest were larger. Since chemical analyses showed that levels of calcium and phosphorus were unchanged with age, our study suggests that during aging the existing bone mineral is reorganized and forms larger aggregates. Given the mechanical disadvantage of large-grained structures (decreased material strength), the observed nanostructural differences contribute to our understanding of the increased fragility of the lateral femoral neck in aged females. Moreover, increasing data on mineral grains in natural bone is essential for advancing calcium-phosphate ceramics for bone tissue replacement.

Atomic force microscopy characterization of the external cortical bone surface in young and elderly women: potential nanostructural traces of periosteal bone apposition during aging.

On the basis of the suggestion that bone nanostructure bears “tissue age” information and may reflect surface deposition/modification processes, we performed nanoscale characterization of the external cortical bone surface at the femoral neck in women using atomic force microscopy (AFM). The specific aims were to assess age-related differences in bone nanostructure and explore the existence of nanostructural traces of potential bone apposition at this surface. Our findings revealed that the external cortical surface represents a continuous phase composed of densely packed mineral grains. Although the grains varied in size and shape, there was a domination of small grains indicative of freshly deposited bone (mean grain size: young, 35 nm; old, 37 nm; p > 0.05). Advanced quantitative analysis of surface morphological patterns revealed comparable roughness and complexity of the surface, suggesting a similar rate of mineral particle deposition at the surface in both groups. Calcium/phosphorus ratio, a measure of bone tissue age, was within the same range in both groups. In summary, our AFM analyses showed consistent nanostructural and compositional bone features, suggesting existence of new bone at the periosteal bone surface in both young and elderly women. Considering observed age-related increase in the neck diameter, AFM findings may support the theory of continuous bone apposition at the periosteal surface.