Methods: A Comparative Analysis of Radiography, Microcomputed Tomography, and Histology for Bone Tissue Engineering

Abstract

This study focused on the assessment of radiography, microcomputed tomography, and histology for the evaluation of bone formation in a 15.0-mm defect in the rabbit radius after the implantation of a tissue-engineered construct. Radiography was found to be useful as a noninvasive method for obtaining images of calcified tissue throughout the time course of the experiment. With this method, however, image quality was low, making it difficult to obtain precise information about the location and quantity of the bone formed. Microcomputed tomography was used to create three-dimensional reconstructions of the bone (25-microm resolution). These reconstructions allowed for greater spatial resolution than the radiography, but did not allow for imaging of the implanted scaffold material or the surrounding, nonmineralized tissue. To visualize all materials within the defect area at the cellular level, histology was used. Histological analysis, however, is a destructive technique that did not allow for any further analysis of the samples. Each technique examined here has its own advantages and limitations, but each yields unique information regarding bone regeneration. It is only through the use of all three techniques that complete characterization of the bone growth and tissue/construct responses after implantation in vivo.

Kinetics of in vivo bone deposition by bone marrow stromal cells within a resorbable porous calcium phosphate scaffold: An X‐ray computed microtomography study

“…Conversely, radiography is a non-invasive method to obtain images throughout the time course of clinical experiments, but is suffering from the low image quality, due to a projection of a 3D structure on a plane thus making it difficult to attain accurate information. Unlike to the above methods, microCT employs conventional X-rays, which can achieve a resolution of about 10 mm, when commercial instruments are used, and up to 1 mm, when Synchrotron Radiation is used (Hedberg et al, 2005). Based on the fact that new bone, fibrous tissue and ceramic scaffolds present different coefficients of absorption, this method permits to separate their 3D structures and to obtain the corresponding quantitative data.…”

Three-Dimensional Characterization of Tissue-Engineered Constructs by Contrast-Enhanced Nanofocus Computed Tomography

“…Currently, standard techniques such as histological sectioning and Live/Dead viability/cytotoxicity staining show limited potential as quality controls for TE constructs as these techniques only allow assessment of tissue distribution in two dimensions, with loss of information and with limited depth resolution, while being destructive in nature. [1][2][3][4] Techniques such as confocal microscopy may offer a potential for three-dimensional (3D) visualization, however, again a limited depth resolution (*300 mm) hinders their performance when larger TE constructs need to be analyzed. 5 Recent advances in 3D imaging techniques and image analysis strategies have demonstrated the potential of addressing some of the shortfalls of these currently applied methods for accurate TE construct analysis.…”

Effect of βTricalcium Phosphate Nanoparticles Additions on the Properties of Gelatin-Chitosan Scaffolds

“…Micro-CT is a very useful, non-destructive tool that provides the architectural analysis of a scaffold used for tissue engineering before implantation [38]. Figures 5a and 5b) shows the representative photographs and 2D images of GCT scaffold using micro CT.…”