Temporal analysis of embryonic bone development using an innovative organotypic bone culture model - application of micro-computed tomography

Abstract

Event Abstract Back to Event Temporal analysis of embryonic bone development using an innovative organotypic bone culture model - application of micro-computed tomography JM Kanczler1*, EL Smith1, CA Roberts1 and ROC Oreffo1 1 University of Southampto, Bone & Joint Research Group, United Kingdom Significant challenges exist for constructing complex tissues, such as bone, that can only be informed by a thorough understanding of the developing tissue environment. Understanding skeletal development of embryonic bone will offer new paradigms for bone augmentation. The organ culture of bone allows for skeletal cells to remain within their extracellular matrix structure while the external environment can be manipulated. The present study has investigated the development of embryonic bone and the effects of different culture conditions on chick femora development in a novel three-dimensional organotypic culture system using micro-computed tomography (microCT).{BR}The length and structure of isolated embryonic chick femurs from E10-E17 were evaluated using microCT. Additionally, femurs were isolated at E10-E13 and placed in organotypic cultures for 10 days in foetal calf serum free culture medium (basal, chondrogenic or osteogenic conditions). Organotypic femurs were analyzed by microCT and assessed histologically for proteoglycans (alcian blue) and collagen (Sirius red) production.{BR}In the developing embryonic chick, femur length (mm) increased from E10=4.9±0.5 to E17=14.2±0.8. microCT analysis (10micron resolution) demonstrated a 152-fold increase in Bone Volume/Total Volume (BV/TV); a 14-fold increase in Trabecular Thickness (TbTh(mm3); a 296-fold increase in Trabecular Number (TbNo(mm)) and a 280-fold decrease in Trabecular Spacing (TbSp (mm)). The embryonic chick femurs-E11 group demonstrated the most significant changes in microCT bone structural elements over the 10 day organotypic culture either in basal, chondrogenic or osteogenic conditions. {BR}E11 isolated femurs, organotypically cultured for 10 days, increased their length compared to control femurs by 3.1mm (Basal); 2.0mm (Chondrogenic) and 2.4mm (Osteogenic) respectively. This was reflected in significant increases (***P<0.001) in microCT bone parameters; (BV/TV), TbTh(mm3), TbNo(mm) and decreased TbSp(mm) in the organotypic culture groups: (BV/TV): Control-E11=0.002±0.001; Basal (d10)=0.009±0.003; Chondrogenic (d10)=0.008±0; Osteogenic (d10)=0.014±0.002; Significant differences were observed in microCT bone parameters (***P<0.001) in the osteogenic organotypic culture groups compared to basal and chondrogenic cultures. However, TbTh(mm3) remained the same across the culture groups.{BR}The current studies demonstrate the efficacy of microCT to interrogate skeletal development. Furthermore, these studies demonstrate the ability to manipulate the developmental window. We believe understanding skeletal developmental biology will underpin and inform the skeletal regenerative process. Keywords: Bones, Bone Research Conference: 2011 joint meeting of the Bone Research Society & the British Orthopaedic Research Society, Cambridge, United Kingdom, 27 Jun - 29 Jun, 2011. Presentation Type: Oral Poster Topic: Abstracts Citation: Kanczler J, Smith E, Roberts C and Oreffo R (2011). Temporal analysis of embryonic bone development using an innovative organotypic bone culture model - application of micro-computed tomography. Front. Endocrinol. Conference Abstract: 2011 joint meeting of the Bone Research Society & the British Orthopaedic Research Society. doi: 10.3389/conf.fendo.2011.02.00032 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 30 Sep 2011; Published Online: 30 Sep 2011. * Correspondence: Dr. JM Kanczler, University of Southampto, Bone & Joint Research Group, Southampton, United Kingdom, jk9@soton.ac.uk Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers JM Kanczler EL Smith CA Roberts ROC Oreffo Google JM Kanczler EL Smith CA Roberts ROC Oreffo Google Scholar JM Kanczler EL Smith CA Roberts ROC Oreffo PubMed JM Kanczler EL Smith CA Roberts ROC Oreffo Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.