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Remodelling of human bone on the chorioallantoic membrane of the chicken egg: De novo bone formation and resorption

Traditionally used as an angiogenic assay, the chorioallantoic membrane (CAM) assay of the chick embryo offers significant potential as an model for xenograft organ culture. Viable human bone can be cultivated on the CAM and increases in bone volume are evident; however, it remains unclear by what m... Full description

Journal Title: Journal of Tissue Engineering and Regenerative Medicine August 2018, Vol.12(8), pp.1877-1890
Main Author: Moreno‐Jiménez, Inés
Other Authors: Lanham, Stuart A. , Kanczler, Janos M. , Hulsart‐Billstrom, Gry , Evans, Nicholas D. , Oreffo, Richard O. C.
Format: Electronic Article Electronic Article
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ID: ISSN: 1932-6254 ; E-ISSN: 1932-7005 ; DOI: 10.1002/term.2711
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recordid: wj10.1002/term.2711
title: Remodelling of human bone on the chorioallantoic membrane of the chicken egg: De novo bone formation and resorption
format: Article
creator:
  • Moreno‐Jiménez, Inés
  • Lanham, Stuart A.
  • Kanczler, Janos M.
  • Hulsart‐Billstrom, Gry
  • Evans, Nicholas D.
  • Oreffo, Richard O. C.
subjects:
  • 3d Registration
  • In Vivo
  • Bmp2
  • Preclinical Model
  • Chorioallantoic Membrane Cam Assay
  • Human Bone
  • Micro Computed Tomography Μct
  • Tissue Engineering
ispartof: Journal of Tissue Engineering and Regenerative Medicine, August 2018, Vol.12(8), pp.1877-1890
description: Traditionally used as an angiogenic assay, the chorioallantoic membrane (CAM) assay of the chick embryo offers significant potential as an model for xenograft organ culture. Viable human bone can be cultivated on the CAM and increases in bone volume are evident; however, it remains unclear by what mechanism this change occurs and whether this reflects the physiological process of bone remodelling. In this study we tested the hypothesis that CAM‐induced bone remodelling is a consequence of host and graft mediated processes. Bone cylinders harvested from femoral heads post surgery were placed on the CAM of green fluorescent protein (GFP)‐chick embryos for 9 days, followed by micro computed tomography (μCT) and histological analysis. Three‐dimensional registration of consecutive μCT‐scans showed newly mineralised tissue in CAM‐implanted bone cylinders, as well as new osteoid deposition histologically. Immunohistochemistry demonstrated the presence of bone resorption and formation markers (Cathepsin K, SOX9 and RUNX2) co‐localising with GFP staining, expressed by avian cells only. To investigate the role of the human cells in the process of bone formation, decellularised bone cylinders were implanted on the CAM and comparable increases in bone volume were observed, indicating that avian cells were responsible for the bone mineralisation process. Finally, CAM‐implantation of acellular collagen sponges, containing bone morphogenetic protein 2, resulted in the deposition of extracellular matrix and tissue mineralisation. These studies indicate that the CAM can respond to osteogenic stimuli and support formation or resorption of implanted human bone, providing a humanised CAM model for regenerative medicine research and a novel short‐term model for tissue engineering and biomaterial testing.
language:
source:
identifier: ISSN: 1932-6254 ; E-ISSN: 1932-7005 ; DOI: 10.1002/term.2711
fulltext: fulltext
issn:
  • 1932-6254
  • 19326254
  • 1932-7005
  • 19327005
url: Link


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titleRemodelling of human bone on the chorioallantoic membrane of the chicken egg: De novo bone formation and resorption
creatorMoreno‐Jiménez, Inés ; Lanham, Stuart A. ; Kanczler, Janos M. ; Hulsart‐Billstrom, Gry ; Evans, Nicholas D. ; Oreffo, Richard O. C.
ispartofJournal of Tissue Engineering and Regenerative Medicine, August 2018, Vol.12(8), pp.1877-1890
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subject3d Registration ; In Vivo ; Bmp2 ; Preclinical Model ; Chorioallantoic Membrane Cam Assay ; Human Bone ; Micro Computed Tomography Μct ; Tissue Engineering
descriptionTraditionally used as an angiogenic assay, the chorioallantoic membrane (CAM) assay of the chick embryo offers significant potential as an model for xenograft organ culture. Viable human bone can be cultivated on the CAM and increases in bone volume are evident; however, it remains unclear by what mechanism this change occurs and whether this reflects the physiological process of bone remodelling. In this study we tested the hypothesis that CAM‐induced bone remodelling is a consequence of host and graft mediated processes. Bone cylinders harvested from femoral heads post surgery were placed on the CAM of green fluorescent protein (GFP)‐chick embryos for 9 days, followed by micro computed tomography (μCT) and histological analysis. Three‐dimensional registration of consecutive μCT‐scans showed newly mineralised tissue in CAM‐implanted bone cylinders, as well as new osteoid deposition histologically. Immunohistochemistry demonstrated the presence of bone resorption and formation markers (Cathepsin K, SOX9 and RUNX2) co‐localising with GFP staining, expressed by avian cells only. To investigate the role of the human cells in the process of bone formation, decellularised bone cylinders were implanted on the CAM and comparable increases in bone volume were observed, indicating that avian cells were responsible for the bone mineralisation process. Finally, CAM‐implantation of acellular collagen sponges, containing bone morphogenetic protein 2, resulted in the deposition of extracellular matrix and tissue mineralisation. These studies indicate that the CAM can respond to osteogenic stimuli and support formation or resorption of implanted human bone, providing a humanised CAM model for regenerative medicine research and a novel short‐term model for tissue engineering and biomaterial testing.
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titleRemodelling of human bone on the chorioallantoic membrane of the chicken egg: De novo bone formation and resorption
descriptionTraditionally used as an angiogenic assay, the chorioallantoic membrane (CAM) assay of the chick embryo offers significant potential as an model for xenograft organ culture. Viable human bone can be cultivated on the CAM and increases in bone volume are evident; however, it remains unclear by what mechanism this change occurs and whether this reflects the physiological process of bone remodelling. In this study we tested the hypothesis that CAM‐induced bone remodelling is a consequence of host and graft mediated processes. Bone cylinders harvested from femoral heads post surgery were placed on the CAM of green fluorescent protein (GFP)‐chick embryos for 9 days, followed by micro computed tomography (μCT) and histological analysis. Three‐dimensional registration of consecutive μCT‐scans showed newly mineralised tissue in CAM‐implanted bone cylinders, as well as new osteoid deposition histologically. Immunohistochemistry demonstrated the presence of bone resorption and formation markers (Cathepsin K, SOX9 and RUNX2) co‐localising with GFP staining, expressed by avian cells only. To investigate the role of the human cells in the process of bone formation, decellularised bone cylinders were implanted on the CAM and comparable increases in bone volume were observed, indicating that avian cells were responsible for the bone mineralisation process. Finally, CAM‐implantation of acellular collagen sponges, containing bone morphogenetic protein 2, resulted in the deposition of extracellular matrix and tissue mineralisation. These studies indicate that the CAM can respond to osteogenic stimuli and support formation or resorption of implanted human bone, providing a humanised CAM model for regenerative medicine research and a novel short‐term model for tissue engineering and biomaterial testing.
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titleRemodelling of human bone on the chorioallantoic membrane of the chicken egg: De novo bone formation and resorption
authorMoreno‐Jiménez, Inés ; Lanham, Stuart A. ; Kanczler, Janos M. ; Hulsart‐Billstrom, Gry ; Evans, Nicholas D. ; Oreffo, Richard O. C.
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abstractTraditionally used as an angiogenic assay, the chorioallantoic membrane (CAM) assay of the chick embryo offers significant potential as an model for xenograft organ culture. Viable human bone can be cultivated on the CAM and increases in bone volume are evident; however, it remains unclear by what mechanism this change occurs and whether this reflects the physiological process of bone remodelling. In this study we tested the hypothesis that CAM‐induced bone remodelling is a consequence of host and graft mediated processes. Bone cylinders harvested from femoral heads post surgery were placed on the CAM of green fluorescent protein (GFP)‐chick embryos for 9 days, followed by micro computed tomography (μCT) and histological analysis. Three‐dimensional registration of consecutive μCT‐scans showed newly mineralised tissue in CAM‐implanted bone cylinders, as well as new osteoid deposition histologically. Immunohistochemistry demonstrated the presence of bone resorption and formation markers (Cathepsin K, SOX9 and RUNX2) co‐localising with GFP staining, expressed by avian cells only. To investigate the role of the human cells in the process of bone formation, decellularised bone cylinders were implanted on the CAM and comparable increases in bone volume were observed, indicating that avian cells were responsible for the bone mineralisation process. Finally, CAM‐implantation of acellular collagen sponges, containing bone morphogenetic protein 2, resulted in the deposition of extracellular matrix and tissue mineralisation. These studies indicate that the CAM can respond to osteogenic stimuli and support formation or resorption of implanted human bone, providing a humanised CAM model for regenerative medicine research and a novel short‐term model for tissue engineering and biomaterial testing.
doi10.1002/term.2711
pages1877-1890
orcidid0000-0002-0717-3671
date2018-08