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

Remodelling of human bone on the chorioallantoic membrane (CAM) of the chicken egg: De novo bone formation and resorption
Remodelling of human bone on the chorioallantoic membrane (CAM) 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 in vivo 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 GFP‐chick embryos for 9 days, followed by micro‐computed tomography (μCT) and histologically. 3D 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 in 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 BMP2, resulted in the deposition of extracellular matrix and tissue mineralisation. These studies indicate that the CAM can respond to osteogenic stimuli and support formation/resorption of implanted human bone; providing a humanised CAM model for regenerative medicine research and a novel short‐term in vivo model for tissue engineering and biomaterial testing.
1932-6254
Oreffo, Richard
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Moreno-Jimenez, Ines
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Lanham, Stuart
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Kanczler, Janos
eb8db9ff-a038-475f-9030-48eef2b0559c
Hulsart Billstrom, Signe Gry
41799212-48c4-4791-9a24-9090a77ba6cc
Evans, Nicholas
06a05c97-bfed-4abb-9244-34ec9f4b4b95
Oreffo, Richard
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Moreno-Jimenez, Ines
0b3c9f73-4124-461e-9cd5-2029ef3eb56c
Lanham, Stuart
28fdbbef-e3b6-4fdf-bd0f-4968eeb614d6
Kanczler, Janos
eb8db9ff-a038-475f-9030-48eef2b0559c
Hulsart Billstrom, Signe Gry
41799212-48c4-4791-9a24-9090a77ba6cc
Evans, Nicholas
06a05c97-bfed-4abb-9244-34ec9f4b4b95

Oreffo, Richard, Moreno-Jimenez, Ines, Lanham, Stuart, Kanczler, Janos, Hulsart Billstrom, Signe Gry and Evans, Nicholas (2018) Remodelling of human bone on the chorioallantoic membrane (CAM) of the chicken egg: De novo bone formation and resorption. Journal of Tissue Engineering and Regenerative Medicine. (doi:10.1002/term.2711).

Record type: Article

Abstract

Traditionally used as an angiogenic assay, the chorioallantoic membrane (CAM) assay of the chick embryo offers significant potential as an in vivo 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 GFP‐chick embryos for 9 days, followed by micro‐computed tomography (μCT) and histologically. 3D 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 in 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 BMP2, resulted in the deposition of extracellular matrix and tissue mineralisation. These studies indicate that the CAM can respond to osteogenic stimuli and support formation/resorption of implanted human bone; providing a humanised CAM model for regenerative medicine research and a novel short‐term in vivo model for tissue engineering and biomaterial testing.

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Moreno et al. - 2018 - Human bone on CAM-De novo bone formation resorption JTERM
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Accepted/In Press date: 12 June 2018
e-pub ahead of print date: 12 June 2018

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Local EPrints ID: 421553
URI: http://eprints.soton.ac.uk/id/eprint/421553
ISSN: 1932-6254
PURE UUID: c65e0542-94c7-4f87-8e77-ac230a017f12
ORCID for Richard Oreffo: ORCID iD orcid.org/0000-0001-5995-6726
ORCID for Stuart Lanham: ORCID iD orcid.org/0000-0002-4516-264X
ORCID for Janos Kanczler: ORCID iD orcid.org/0000-0001-7249-0414

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Date deposited: 15 Jun 2018 16:30
Last modified: 07 Oct 2020 05:35

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Contributors

Author: Richard Oreffo ORCID iD
Author: Ines Moreno-Jimenez
Author: Stuart Lanham ORCID iD
Author: Janos Kanczler ORCID iD
Author: Signe Gry Hulsart Billstrom
Author: Nicholas Evans

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