Bone cell network viability: novel 3D mapping of cancellous osteocyte interconnection in the ageing human femoral head

Abstract

Event Abstract Back to Event Bone cell network viability: novel 3D mapping of cancellous osteocyte interconnection in the ageing human femoral head PE Garner1*, PE Garner2, RK Wilcox2 and JE Aaron1 1 University of Leeds, IMSB, FBS, United Kingdom 2 University of Leeds, IMBE, United Kingdom Osteocytes are the most abundant bone cell (90-95%) and in normal tissue each may remain viable for many years. Despite entombment in calcified matrix their extensive processes form a pervasive syncytium, similar to that of neurones in the central nervous system, and whereby no part of bone is more than a few microns from an osteocyte. This suggests that they are not simply passive, but that they have a role in mechanotransduction, influencing remodelling and self-repair and functioning as the skeletal mechanostat (Frost 2003). To gain insight into the potential morphological variability of the network with reference to skeletal location and to associated stress input, a novel method has been developed combining undecalcified histology, confocal microscopy and image analysis software that enables the quantitative characterisation of the major 3D network features. The method has been subsequently applied to quartered, ageing human femoral heads (a particularly vulnerable skeletal site) to identify any regional differences (e.g. relative to tendon insertion proximity versus compression sites) and also to compare the network in contrasting conditions of traditionally high stress (osteoarthritis) and low stress (osteoporosis). Bone segments underwent en-bloc staining in the fluorochrome 0.1% calcein for 3 days under vacuum before embedding in methylmethacrylate resin. Thick (300 microns) undecalcified slices were cut and scanned and image z-stacks (approx. 200-250 x-y images 0.3 microns apart) captured using a laser scanning confocal microscope with a krypton/argon laser, a 500nm excitation filter and 700nm emission filter. Individual x-y 2D Tiff images were imported into Simpleware (analytical software) to generate a complementary pair of 3D binary masks representative of either the osteocyte cell bodies alone or of the extensive processes alone. Corresponding in-house code (Matlab, Mathworks, USA) was written to quantitate the complementary paired masked aspects including the volume of the cell body component, the number of processes/cell, their length, interconnection and anisotropy. The objective outcome will augment previous subjective appraisal of the syncitium, leading to a more precise understanding of its capacity for musculoskeletal exchange and bone mass determination in ageing and disease. 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: Poster Topic: Abstracts Citation: Garner P, Garner P, Wilcox R and Aaron J (2011). Bone cell network viability: novel 3D mapping of cancellous osteocyte interconnection in the ageing human femoral head. Front. Endocrinol. Conference Abstract: 2011 joint meeting of the Bone Research Society & the British Orthopaedic Research Society. doi: 10.3389/conf.fendo.2011.02.00020 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: Prof. PE Garner, University of Leeds, IMSB, FBS, United Kingdom, bms2peg@leeds.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 PE Garner PE Garner RK Wilcox JE Aaron Google PE Garner PE Garner RK Wilcox JE Aaron Google Scholar PE Garner PE Garner RK Wilcox JE Aaron PubMed PE Garner PE Garner RK Wilcox JE Aaron 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.