Haviva M. Goldman

Lamellar Bone is an Incremental Tissue Reconciling Enamel Rhythms, Body Size, and Organismal Life History

Mammalian enamel formation is periodic, including fluctuations attributable to the daily biological clock as well as longer-period oscillations that enigmatically correlate with body mass. Because the scaling of bone mass to body mass is an axiom of vertebrate hard tissue biology, we consider that long-period enamel formation rhythms may reflect corresponding and heretofore unrecognized rhythms in bone growth. The principal aim of this study is to seek a rhythm in bone growth demonstrably related to enamel oscillatory development. Our analytical approach is based in morphology, using a variety of hard tissue microscopy techniques. We first ascertain the relationship among long-period enamel rhythms, the striae of Retzius, and body mass using a large sample of mammalian taxa. In addition, we test whether osteocyte lacuna density (a surrogate for rates of cell proliferation) in bone is correlated with mammalian body mass. Finally, using fluorescently labeled developing bone tissues, we investigate whether the bone lamella, a fundamental microanatomical unit of bone, relates to rhythmic enamel growth increments. Our results confirm a positive correlation between long-period enamel rhythms and body mass and a negative correlation between osteocyte density and body mass. We also confirm that lamellar bone is an incremental tissue, one lamella formed in the species-specific time dependency of striae of Retzius formation. We conclude by contextualizing our morphological research with a current understanding of autonomic regulatory control of the skeleton and body mass, suggesting a central contribution to the coordination of organismal life history and body mass.

Genetic Variation in Bone Growth Patterns Defines Adult Mouse Bone Fragility

Femoral morphology and composition were determined for three inbred mouse strains between ages E18.5 and 1 year. Genotype‐specific variation in postnatal, pubertal, and postpubertal growth patterns and mineral accrual explained differences in adult bone trait combinations and thus bone fragility.

Ontogenetic Patterning of Cortical Bone Microstructure and Geometry at the Human Mid‐Shaft Femur

The bone growth process has long‐lasting effects on adult bone structure and mechanical adaptation, yet the tissue level dynamics of growth are poorly studied. The specific aims of this study were to (1) quantify changes in bone size and shape through ontogeny, (2) describe the distribution of tissue types and patterns of cortical drift and expansion through ontogeny, and (3) explore relationships between cortical drift and ontogenetic variation geometric size and shape. The study utilized 14 juvenile (ages 2–19) mid‐shaft femur blocks removed at autopsy from individuals who died suddenly. Eighty‐μm‐thick sections were imaged using polarized and brightfield microscopy. For descriptive purposes the sample was divided into five age groups. Features of collagen fiber matrix orientation, vascularity (e.g., pore orientation and density), and osteocyte lacunar density and shape were used to classify primary and secondary tissue types in LM images. This information, combined with evaluation of resorptive versus depositional bone surfaces, was used to identify cortical drift direction. A pattern of posterior and medial drift was identified at the mid‐shaft femur in the toddler years. The drift pattern shifts antero‐laterally in late childhood, predating the appearance of a more adult‐like geometry. On the basis of the presence of transitional fibrolamellar bone complex, growth is more rapid during the toddler years and peri‐puberty, and slower in early to late childhood and in later adolescence. Extensive variability in histological and geometric organization typifies the sample, particularly beginning in late childhood. The potential implications of this variability for adult fracture risk warrant further study. Anat Rec, 2009.

Measuring the dynamic mechanical response of hydrated mouse bone by nanoindentation

This study demonstrates a novel approach to characterizing hydrated bones viscoelastic behavior at lamellar length scales using dynamic indentation techniques. We studied the submicron-level viscoelastic response of bone tissue from two different inbred mouse strains, A/J and B6, with known differences in whole bone and tissue-level mechanical properties. Our results show that bone having a higher collagen content or a lower mineral-to-matrix ratio demonstrates a trend towards a larger viscoelastic response. When normalized for anatomical location relative to biological growth patterns in the antero-medial (AM) cortex, bone tissue from B6 femora, known to have a lower mineral-to-matrix ratio, is shown to exhibit a significantly higher viscoelastic response compared to A/J tissue. Newer bone regions with a higher collagen content (closer to the endosteal edge of the AM cortex) showed a trend towards a larger viscoelastic response. Our study demonstrates the feasibility of this technique for analyzing local composition-property relationships in bone. Further, this technique of viscoelastic nanoindentation mapping of the bone surface at these submicron length scales is shown to be highly advantageous in studying subsurface features, such as porosity, of wet hydrated biological specimens, which are difficult to identify using other methods.

Biological constraints that limit compensation of a common skeletal trait variant lead to inequivalence of tibial function among healthy young adults.

Having a better understanding of how complex systems like bone compensate for the natural variation in bone width to establish mechanical function will benefit efforts to identify traits contributing to fracture risk. Using a collection of pQCT images of the tibial diaphysis from 696 young adult women and men, we tested the hypothesis that bone cells cannot surmount the nonlinear relationship between bone width and whole bone stiffness to establish functional equivalence across a healthy population. Intrinsic cellular constraints limited the degree of compensation, leading to functional inequivalence relative to robustness, with slender tibias being as much as two to three times less stiff relative to body size compared with robust tibias. Using Path Analysis, we identified a network of compensatory trait interactions that explained 79% of the variation in whole‐bone bending stiffness. Although slender tibias had significantly less cortical area relative to body size compared with robust tibias, it was the limited range in tissue modulus that was largely responsible for the functional inequivalence. Bone cells coordinately modulated mineralization as well as the cortical porosity associated with internal bone multicellular units (BMU)‐based remodeling to adjust tissue modulus to compensate for robustness. Although anecdotal evidence suggests that functional inequivalence is tolerated under normal loading conditions, our concern is that the functional deficit of slender tibias may contribute to fracture susceptibility under extreme loading conditions, such as intense exercise during military training or falls in the elderly. Thus, we show the natural variation in bone robustness was associated with predictable functional deficits that were attributable to cellular constraints limiting the amount of compensation permissible in human long bone. Whether these cellular constraints can be circumvented prophylactically to better equilibrate function among individuals remains to be determined.

Relationships among microstructural properties of bone at the human midshaft femur.

Mineralization density and collagen fibre orientation are two aspects of a bones microstructural organization that influence its mechanical properties. Previous studies by our group have demonstrated a distinctly non‐random, though highly variable, spatial distribution of these two variables in the human femoral cortex. In this study of 37 specimens, these variables are examined relative to one another in order to determine whether regions of bone demonstrating higher or lower mineralization density also demonstrate a prevalence of either transversely or longitudinally oriented collagen fibres. An analysis of rank‐transformed collagen fibre orientation (as determined by circularly polarized light) and mineralization density (as determined by backscattered electron microscopy) data sets demonstrated that areas of low mineralization density (predominantly in the anterior‐lateral cortex) tended to correspond to regions of higher proportions of longitudinally oriented collagen fibres. Conversely, areas of higher mineralization density (postero‐medially) tended to correspond to regions of higher proportions of transversely oriented collagen fibres. High variability in the sample led to generally low correlations between the two data sets, however. A second analysis focused only on the orientation of collagen fibres within poorly mineralized bone (representing bone that was newly formed). This analysis demonstrated a lower proportion of transverse collagen fibres in newly formed bone with age, along with some significant regional differences in the prevalence of collagen fibres of either orientation. Again high variability characterized the sample. These results are discussed relative to the hypothesized forces experienced at the midshaft femur.

Assessment of lamellar level properties in mouse bone utilizing a novel spherical nanoindentation data analysis method

In this work, we demonstrate the viability of using our recently developed data analysis procedures for spherical nanoindentation in conjunction with Raman spectroscopy for studying lamellar-level correlations between the local composition and local mechanical properties in mouse bone. Our methodologies allow us to convert the raw load-displacement datasets to much more meaningful indentation stress-strain curves that accurately capture the loading and unloading elastic moduli, the indentation yield points, as well as the post-yield characteristics in the tested samples. Using samples of two different inbred mouse strains, A/J and C57BL/6J (B6), we successfully demonstrate the correlations between the mechanical information obtained from spherical nanoindentation measurements to the local composition measured using Raman spectroscopy. In particular, we observe that a higher mineral-to-matrix ratio correlated well with a higher local modulus and yield strength in all samples. Thus, new bone regions exhibited lower moduli and yield strengths compared to more mature bone. The B6 mice were also found to exhibit lower modulus and yield strength values compared to the more mineralized A/J strain.

Surgical Treatment Options in Patients With Impaired Bone Quality

BackgroundBone quality should play an important role in decision-making for orthopaedic treatment options, implant selection, and affect ultimate surgical outcomes. The development of decision-making tools, currently typified by clinical guidelines, is highly dependent on the precise definition of the term(s) and the appropriate design of basic and clinical studies. This review was performed to determine the extent to which the issue of bone quality has been subjected to this type of process.Questions/purposesWe address the following issues: (1) current methods of clinically assessing bone quality; (2) emerging technologies; (3) how bone quality connects with surgical decision-making and the ultimate surgical outcome; and (4) gaps in knowledge that need to be closed to better characterize bone quality for more relevance to clinical decision-making.MethodsPubMed was used to identify selected papers relevant to our discussion. Additional sources were found using the references cited by identified papers.ResultsBone mineral density remains the most commonly validated clinical reference; however, it has had limited specificity for surgical decision-making. Other structural and geometric measures have not yet received enough study to provide definitive clinical applicability. A major gap remains between the basic research agenda for understanding bone quality and the transfer of these concepts to evidence-based practice.ConclusionsBasic bone quality needs better definition through the systematic study of emerging technologies that offer a more precise clinical characterization of bone. Collaboration between basic scientists and clinicians needs to improve to facilitate the development of key questions for sound clinical studies.

The virtual microscopy database-sharing digital microscope images for research and education: Virtual Microcopy Database

Over the last 20 years, virtual microscopy has become the predominant modus of teaching the structural organization of cells, tissues, and organs, replacing the use of optical microscopes and glass slides in a traditional histology or pathology laboratory setting. Although virtual microscopy image files can easily be duplicated, creating them requires not only quality histological glass slides but also an expensive whole slide microscopic scanner and massive data storage devices. These resources are not available to all educators and researchers, especially at new institutions in developing countries. This leaves many schools without access to virtual microscopy resources. The Virtual Microscopy Database (VMD) is a new resource established to address this problem. It is a virtual image file‐sharing website that allows researchers and educators easy access to a large repository of virtual histology and pathology image files. With the support from the American Association of Anatomists (Bethesda, MD) and MBF Bioscience Inc. (Williston, VT), registration and use of the VMD are currently free of charge. However, the VMD site is restricted to faculty and staff of research and educational institutions. Virtual Microscopy Database users can upload their own collection of virtual slide files, as well as view and download image files for their own non‐profit educational and research purposes that have been deposited by other VMD clients. Anat Sci Educ 11: 510–515.