There is a plethora of calcium phosphate (CaP) scaffolds used as synthetic substitutes to bone grafts. The scaffold performance is often evaluated from the quantity of bone formed within or in direct contact with the scaffold. Micro-computed tomography (µCT) allows three-dimensional evaluation of bone formation inside scaffolds. However, the almost identical x-ray attenuation of CaP and bone obtrude the separation of these phases in µCT images. Commonly, segmentation of bone in µCT images is based on gray scale intensity, with manually determined global thresholds. However, image analysis methods, and methods for manual thresholding in particular, lack standardization and may consequently suffer from subjectivity. The aim of the present study was to provide a methodological framework for addressing these issues. Bone formation in two types of CaP scaffold architectures (foamed and robocast), obtained from a larger animal study (a 12 week canine animal model) was evaluated by µCT. In addition, cross-sectional scanning electron microscopy (SEM) images were acquired as references to determine thresholds and to validate the result. µCT datasets were registered to the corresponding SEM reference. Global thresholds were then determined by quantitatively correlating the different area fractions in the µCT image, towards the area fractions in the corresponding SEM image. For comparison, area fractions were also quantified using global thresholds determined manually by two different approaches. In the validation the manually determined thresholds resulted in large average errors in area fraction (up to 17%), whereas for the evaluation using SEM references, the errors were estimated to be less than 3%. Furthermore, it was found that basing the thresholds on one single SEM reference gave lower errors than determining them manually. This study provides an objective, robust and less error prone method to determine global thresholds for the evaluation of bone formation in CaP scaffolds.
Ambrosio, L.; Guarino, V.; Torricelli, P.; Fini, M.; Ginebra, M.P.; Planell, J. A.; Giardino, R. Biomedical materials Vol. 7, num. 2, p. 1-10 DOI: 10.1088/1748-6041/7/2/024113 Data de publicació: 2012 Article en revista
Vlad, M.; del Valle, LJ.; Poeata, I.; Lopez, J.; Torres, R.; Barraco, M.; Fernandez, E. Biomedical materials Vol. 5, num. 2, p. 1-13 DOI: 10.1088/1748-6041/5/2/025006 Data de publicació: 2010-03 Article en revista
Premi del "National University Research Council" del Ministeri d'Educació i Recerca de Romania, a autors romanesos per l'impacte de la seva recerca.
In this study, the cytocompatibility of new ‘iron-modified/alpha-tricalcium phosphate
(IM/α-TCP) and calcium sulfate dihydrate (CSD)’ bone cement (IM/α-TCP/CSD-BC)
intended for spinal applications has been approached. The objective was to investigate by direct-contact osteoblast-like cell cultures (from 1 to 14 days) the in vitro cell adhesion, proliferation, morphology and cytoskeleton organization of MG-63 cells seeded onto the new cements. The results were as follows: (a) quantitative MTT-assay and scanning electron microscopy (SEM) showed that cell adhesion, proliferation and viability were not affected with time by the presence of iron in the cements; (b) double immunofluorescent labeling of F-actin and α-tubulin showed a dynamic interaction between the cell and its porous substrates
sustaining the locomotion phenomenon on the cements’ surface, which favored the
colonization, and confirming the biocompatibility of the experimental cements; (c) SEM-cell morphology and cytoskeleton observations also evidenced that MG-63 cells were able to adhere, to spread and to attain normal morphology on the new IM/α-TCP/CSD-BC which offered favorable substratum properties for osteoblast-like cells proliferation and
differentiation in vitro. The results showed that these new iron-modified cement-like
biomaterials have cytocompatible features of interest not only as possible spinal cancellous bone replacement biomaterial but also as bone tissue engineering scaffolds.
316L Stainless steel is one of the most used metallic material in orthopedical prosthesis, osteosinthesis plates, and cardiovascular stents. One of the main problems this material presents is the nickel and chromium release, specially the Ni ion release that provokes allergy in a high number of patients. Recently, experimental applications in vitro and in vivo seem to indicate that the thickness of the nature oxide of the stainless steel results in very strong reinforcement of the biological response and reduce the ion release due to the thicker surface oxide. It is possible to grow the natural chromium oxide layer by electrolytic method such anodization. In this study, two main anodization methods to grow chromium oxide on the 316L stainless steel have been evaluated. Nickel and Chromium ions release in human blood at 37 degrees C were detected at times of 1, 6, 11, and 15 days by means of atomic absorption in a graphite furnace (GAAF). Moreover, cytocompatibility tests were carried out. Perfusion experiments were performed to evaluate morphometrically platelet interaction with the material and to explore the potential thrombogenicity. The results showed a good cytocompatibility between the material and the osteoblast-like cells. However, these anodization methods released between 2 and 10 times more nickel and chromium than the original stainless steel, depending on the method used. Besides, anodized samples shown an increase of the percentage of surface covered by platelets. Consequently, the anodization methods studied do not improve the long-term behavior of the stainless steel for its application as cardiovascular stents.