Morphometric analysis of 70 bone biopsies was done in parallel by µCT and histomorphometry. µCT provided higher results for trabecular thickness and separation because of the 3D shape of these anatomical objects.Introduction: Bone histomorphometry is used to explore the various metabolic bone diseases. The technique is done on microscopic 2D sections, and several methods have been proposed to extrapolate 2D measurements to the 3D dimension. X-ray CT is a recently developed imaging tool to appreciate 3D architecture. Recently the use of 2D histomorphometric measurements have been shown to provide discordant results compared with 3D values obtained directly. Material and Methods: Seventy human bone biopsies were removed from patients presenting with metabolic bone diseases. Complete bone biopsies were examined by CT. Bone volume (BV/TV), Tb.Th, and Tb.Sp were measured on the 3D models. Tb.Th and Tb.Sp were measured by a method based on the sphere algorithm. In addition, six images were resliced and transferred to an image analyzer: bone volume and trabecular characteristics were measured after thresholding of the images. Bone cores were embedded undecalcified; histological sections were prepared and measured by routine histomorphometric methods providing another set of values for bone volume and trabecular characteristics. Comparison between the different methods was done by using regression analysis, Bland-Altman, Passing-Bablock, and Mountain plots. Results: Correlations between all parameters were highly significant, but CT overestimated bone volume. The osteoid volume had no influence in this series. Overestimation may have been caused by a double threshold used in CT, giving trabecular boundaries less well defined than on histological sections. Correlations between Tb.Th and Tb.Sp values obtained by 3D or 2D measurements were lower, and 3D analysis always overestimated thickness by ∼50%. These increases could be attributed to the 3D shape of the object because the number of nodes and the size of the marrow cavities were correlated with 3D values. Conclusion:In clinical practice, CT seems to be an interesting method providing reliable morphometric results in less time than conventional histomorphometry. The correlation coefficient is not sufficient to study the agreement between techniques in histomorphometry. The architectural descriptors are influenced by the algorithms used in 3D.
Poly(2-hydroxyethyl methacrylate) (pHEMA) has potentially broad biomedical applications: it is biocompatible and has a hardness comparable to bone when bulk polymerized. Porous biomaterials allow bone integration to be increased, especially when the pores are interconnected. In this study, three types of porogens (sugar fibers, sucrose crystals, and urea beads) have been used to prepare macroporous pHEMA. The pore volume and interconnectivity parameters of the porosity were measured by X-ray microtomography and image analysis. Sucrose crystals, having a high volumetric mass, gave large pores that were located on the block sides. Urea beads and sugar fibers provided pores with the same star volume (2.65 +/- 0.46 mm3 and 2.48 +/- 0.52 mm3, respectively) but which differed in interconnectivity index, fractal dimension, and Euler-Poincarés number. Urea beads caused non-connected porosity, while sugar fibers created a dense labyrinth within the polymer. Interconnectivity was proved by carrying out surface treatment of the pHEMA (carboxymethylation in water), followed by von Kossà staining, which detected the carboxylic groups. Carboxymethylated surfaces were observed on the sides of the blocks and on the opened or interconnected pores. The disconnected pores were unstained. Macroporous polymers can be prepared with water-soluble porogens. X-ray microtomography appears a useful tool to measure porosity and interconnectedness.
Commercial ß-tricalcium phosphate (ß-TCP) is commercialy available in granules manufactured by sintering of powders. We have evaluated the different steps of the manufacturing process of ß-TCP ceramics granules prepared from blocks obtained with the polyurethane foam technology. Three types of slurry were prepared with 10, 15 and 25 g of ß-TCP per gram of polyurethane foam. Analysis was done by scanning electron microscopy, EDX, Raman spectroscopy and microcomputed tomography combined with image analysis. A special algorithm was used to identify the internal microporosity (created by the calcination of the foam) from the internal macroporosity due to the spatial repartition of the material. The low ß-TCP dosages readily infiltrated the foam and the slurry was deposited along the polymer rods. On the contrary, the highest concentration produced inhomogeneous infiltrated blocks and foam cavities appeared completely filled in some areas. 2D microcomputed sections and reconstructed 3D models evidenced this phenomenon and the frequency distribution of the thickness and separation of material trabeculae confirmed the heterogeneity of the distribution. When crushed, blocks prepared with the 25 g slurry provided the largest and irregular granulates.
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