Janardhan Yerramshetty

Digital tomosynthesis (DTS) provides slice images of an object using conventional radiographic methods with high in-plane resolution. The objective of this study was to explore the potential of DTS for describing microstructural, stiffness and stress distribution properties of vertebral cancellous bone. Forty vertebrae (T6, T8, T11, and L3) from 10 cadavers (63-90 years) were scanned using microCT and DTS. Anisotropy (μCT.DA), and the specimen-average and standard deviation of trabecular bone volume fraction (BV/TV), thickness (Tb.Th), number (Tb.N) and separation (Tb.Sp) were obtained using stereology. Apparent modulus (EFEM), and the magnitude (VMExp/σapp) and variability (VMCV) of trabecular stresses were calculated using microCT-based finite element modeling. Mean intercept length, line fraction deviation and fractal parameters were obtained from coronal DTS slices, then correlated with stereological and finite element parameters using linear regression models. Twenty-one DTS parameters (out of 27) correlated to BV/TV, Tb.Th, Tb.N, Tb.Sp and/or μCT.DA (p<0.0001-p<0.05). DTS parameters increased the explained variability in EFEM and VMCV (by 9-11% and 13-19%, respectively; p<0.0001-p<0.04) over that explained by BV/TV. In conclusion, DTS has potential for quantitative assessment of cancellous bone and may be used as a modality complementary to those measuring bone mass for assessing spinal fracture risk.

The lack of accuracy in the prediction of vertebral fracture risk from average density measurements, all external factors being equal, may be because bone mineral density (BMD) is less than a perfect surrogate for bone strength but also because strength alone may not be sufficient to fully characterize the structural failure of a vertebra. Apart from bone quantity, organization of micro-architecture would have a role in governing mechanical properties of vertebrae. An earlier study indicated that within-vertebra variability, measured as the coefficient of variation (COV) of bone volume fraction (BV/TV) or as COV of finite element-estimated apparent modulus (EFE) correlated well with vertebral strength [1]. Therefore, as an extension to our earlier study, we investigated i) whether the relationships of vertebral strength found with COV of BV/TV and COV of EFE could be extended to the COV of other micro-structural parameters and microcomputed tomography-estimated bone mineral density (μCT-BMD) and ii) whether COV of microstructural parameters were associated with structural ductility measures.

Postretrieval analysis of biodegradable polymeric constructs for degradation rates requires correct identification of the degradable polymer, de novo tissue and the confounding presence of a secondary polymer used for embedding. Similarities between the structures of many tissue engineering polymers may make them difficult to distinguish from the polymer used to embed explants prior to histological sectioning. In this study, we assessed the feasibility of a chemical imaging method, Raman microscopy, to discriminate between more than one polymer species. From the perspective of spectroscopy, this is not a straightforward process because of the emergence of multiple peaks, ubiquity of embedding medium, and presence of observations sourcing from points sampled at the interface of two phases. A multivariate K‐means data clustering method was used to discriminate between different polymeric components. The method was able to classify 95% of the observations to the correct category. The remaining data displayed multiple memberships because of (1) the laser spot coinciding with the interfaces of more than one phase or (2) infiltration of histological embedding polymer. Combined with multivariate analysis methods, this technique may prove useful in the future for tissue engineering and biomaterials analysis of degradation rates of, and tissue ingrowth into, polymer scaffolds. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res 2007

Raman spectroscopy provides valuable information on the physicochemical properties of hard tissues. While the technique can analyze tissues in their native state, analysis of fixed, embedded, and sectioned specimens may be necessary on certain occasions. The information on the effects of fixatives and embedding media on Raman spectral properties is limited. We examined the effect of ethanol and glycerol as fixatives and a variety of embedding media (Araldite, Eponate, Technovit, glycol methacrylate, polymethyl methacrylate, and LR white) on Raman spectral properties (mineralization, crystallinity, and carbonation) measured from the cortical bone of mouse humeri. Humeri were fixed in ethanol or glycerol, followed by embedding in one of the media. Nonfixed, freeze-dried, and fixed but not embedded sections were also examined. Periosteal, endosteal, and midosteal regions of the intracortical envelope were analyzed. Raman spectra of fixative solutions and embedding media were also recorded separately in order to examine the specifics of overlap between spectra. We found significant effects of fixation, embedding, and anatomical location on Raman spectral properties. The interference of ethanol with tissue seemed to be relatively less pronounced than that of glycerol. However, there was no single combination of fixation and embedding that left Raman spectral parameters unaltered. We conclude that careful selection of a fixation and embedding combination should be made based on the parameter of interest and the type of tissue. It may be necessary to process multiple samples from the tissue, each using a combination appropriate for the Raman parameter in question.