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      Heidelberg-mCT-Analyzer: a novel method for standardized microcomputed-tomography-guided evaluation of scaffold properties in bone and tissue research

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          Abstract

          Bone tissue engineering and bone scaffold development represent two challenging fields in orthopaedic research. Micro-computed tomography (mCT) allows non-invasive measurement of these scaffolds’ properties in vivo. However, the lack of standardized mCT analysis protocols and, therefore, the protocols’ user-dependency make interpretation of the reported results difficult. To overcome these issues in scaffold research, we introduce the Heidelberg-mCT-Analyzer. For evaluation of our technique, we built 10 bone-inducing scaffolds, which underwent mCT acquisition before ectopic implantation (T0) in mice, and at explantation eight weeks thereafter (T1). The scaffolds’ three-dimensional reconstructions were automatically segmented using fuzzy clustering with fully automatic level-setting. The scaffold itself and its pores were then evaluated for T0 and T1. Analysing the scaffolds’ characteristic parameter set with our quantification method showed bone formation over time. We were able to demonstrate that our algorithm obtained the same results for basic scaffold parameters (e.g. scaffold volume, pore number and pore volume) as other established analysis methods. Furthermore, our algorithm was able to analyse more complex parameters, such as pore size range, tissue mineral density and scaffold surface. Our imaging and post-processing strategy enables standardized and user-independent analysis of scaffold properties, and therefore is able to improve the quantitative evaluations of scaffold-associated bone tissue-engineering projects.

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          Most cited references22

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          Potential of ceramic materials as permanently implantable skeletal prostheses.

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            Donor variation and loss of multipotency during in vitro expansion of human mesenchymal stem cells for bone tissue engineering.

            The use of multipotent human mesenchymal stem cells (hMSCs) for tissue engineering has been a subject of extensive research. The donor variation in growth, differentiation and in vivo bone forming ability of hMSCs is a bottleneck for standardization of therapeutic protocols. In this study, we isolated and characterized hMSCs from 19 independent donors, aged between 27 and 85 years, and investigated the extent of heterogeneity of the cells and the extent to which hMSCs can be expanded without loosing multipotency. Dexamethasone-induced ALP expression varied between 1.2- and 3.7-fold, but no correlation was found with age, gender, or source of isolation. The cells from donors with a higher percentage of ALP-positive cells in control and dexamethasone-induced groups showed more calcium deposition than cells with lower percentage of ALP positive cells. Despite the variability in osteogenic gene expression among the donors tested, ALP, Collagen type 1, osteocalcin, and S100A4 showed similar trends during the course of osteogenic differentiation. In vitro expansion studies showed that hMSCs can be effectively expanded up to four passages (approximately 10-12 population doublings from a P0 culture) while retaining their multipotency. Our in vivo studies suggest a correlation between in vitro ALP expression and in vivo bone formation. In conclusion, irrespective of age, gender, and source of isolation, cells from all donors showed osteogenic potential. The variability in ALP expression appears to be a result of sampling method and cellular heterogeneity among the donor population.
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              Mechanisms of bone repair and regeneration.

              Bone problems can have a highly deleterious impact on life and society, therefore understanding the mechanisms of bone repair is important. In vivo studies show that bone repair processes in adults resemble normal development of the skeleton during embryogenesis, which can thus be used as a model. In addition, recent studies of skeletal stem cell biology have underlined several crucial molecular and cellular processes in bone formation. Hedgehog, parathyroid hormone-related protein, Wnt, bone morphogenetic proteins and mitogen-activated protein kinases are the main molecular players, and osteoclasts and mesenchymal stem cells are the main cells involved in these processes. However, questions remain regarding the precise mechanisms of bone formation, how the different molecular processes interact, and the real identity of regenerative cells. Here, we review recent studies of bone regeneration and repair. A better understanding of the underlying mechanisms is expected to facilitate the development of new strategies for improving bone repair.
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                Author and article information

                Journal
                R Soc Open Sci
                R Soc Open Sci
                RSOS
                royopensci
                Royal Society Open Science
                The Royal Society Publishing
                2054-5703
                November 2015
                11 November 2015
                11 November 2015
                : 2
                : 11
                : 150496
                Affiliations
                [1 ]Trauma and Reconstructive Surgery, Center of Orthopedics, Traumatology, and Spinal Cord Injury, Heidelberg University Hospital , Schlierbacher Landstraße 200a, Heidelberg 69118, Germany
                [2 ]HTRG - Heidelberg Trauma Research Group, Schlierbacher Landstraße 200a , Heidelberg 69118, Germany
                [3 ]Clinic of Diagnostic and Interventional Radiology (DIR), Heidelberg University Hospital , Im Neuenheimer Feld 110, Heidelberg 69120, Germany
                Author notes
                Author for correspondence: Arash Moghaddam e-mail: arash.moghaddam@ 123456med.uni-heidelberg.de
                [†]

                These authors contributed equally to this study.

                Author information
                http://orcid.org/0000-0001-9948-4209
                Article
                rsos150496
                10.1098/rsos.150496
                4680623
                7dde9356-b9bd-488c-a5fa-d5476214de9d

                © 2015 The Authors. Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited.

                History
                : 17 September 2015
                : 13 October 2015
                Categories
                1003
                48
                1004
                18
                1009
                124
                Cellular and Molecular Biology
                Research Article
                Custom metadata
                November, 2015

                bone scaffold,bone tissue engineering,micro-computed tomography,scaffold characterization,bmp-7 (bone morphogenetic protein-7),human mesenchymal stem cells

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