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The chorioallantoic membrane assay as an in vivo model for the study of human bone and tissue engineering

The chorioallantoic membrane assay as an in vivo model for the study of human bone and tissue engineering
The chorioallantoic membrane assay as an in vivo model for the study of human bone and tissue engineering
Poor vascularization at the fracture site is one of the main limitations of successful bone healing. To improve vascularisation, new tissue engineering approaches aim to design scaffolds that allow not only osteogenesis but also stable capillary invasion: angiogenesis. Importantly, the efficacy and safety of novel bone tissue engineering constructs needs to be tested in vivo before the constructs can be used clinically, which typically involves invasive surgical procedures in murine hosts.
The present thesis proposes the use the chorioallantoic membrane (CAM) from the developing chick as a less sentient in vivo model to explore human bone healing, and the application of this system as a novel screening platform for biomaterials in tissue engineering. To test this model, human bone cylinders were extracted from fresh human femoral heads, stimulated with an injury using a drill-hole defect, and implanted on the CAM or cultured in vitro for a period of 7-9 days. Micro-computed tomography (µCT) was used to quantify the magnitude of bone volume changes together with histological analysis to follow bone repair.
Bone cylinders were vascularised by the CAM blood vessels and remained viable following implantation throughout the culture period. Histological evaluation showed formation of cell condensations in proximity to avian capillaries (immunocytochemistry positive for Sox9 and Runx2). New extracellular matrix (Collagen type II, X) deposition and osteoclast activity (Cathepsin K) on the CAM-implanted cylinders, correlated with a significant increase in bone volume as determined by µCT analysis (p<0.05) and demonstrated the ability of the human-avian system asa test/screen for biomaterials.
In summary, the current studies demonstrate the potential of the CAM assay as a surrogate blood supply for viable human bone tissue. This human-avian system offers a simple refinement model for animal research and a step towards a humanized in vivo model not only for tissue engineering, but for many other disciplines.
University of Southampton
Moreno Jiménez, Inés
4dbe5fd4-8b2b-48c6-8c48-4833fd7d7d50
Moreno Jiménez, Inés
4dbe5fd4-8b2b-48c6-8c48-4833fd7d7d50
Oreffo, Richard
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Evans, Nicholas
06a05c97-bfed-4abb-9244-34ec9f4b4b95

Moreno Jiménez, Inés (2016) The chorioallantoic membrane assay as an in vivo model for the study of human bone and tissue engineering. University of Southampton, Doctoral Thesis, 278pp.

Record type: Thesis (Doctoral)

Abstract

Poor vascularization at the fracture site is one of the main limitations of successful bone healing. To improve vascularisation, new tissue engineering approaches aim to design scaffolds that allow not only osteogenesis but also stable capillary invasion: angiogenesis. Importantly, the efficacy and safety of novel bone tissue engineering constructs needs to be tested in vivo before the constructs can be used clinically, which typically involves invasive surgical procedures in murine hosts.
The present thesis proposes the use the chorioallantoic membrane (CAM) from the developing chick as a less sentient in vivo model to explore human bone healing, and the application of this system as a novel screening platform for biomaterials in tissue engineering. To test this model, human bone cylinders were extracted from fresh human femoral heads, stimulated with an injury using a drill-hole defect, and implanted on the CAM or cultured in vitro for a period of 7-9 days. Micro-computed tomography (µCT) was used to quantify the magnitude of bone volume changes together with histological analysis to follow bone repair.
Bone cylinders were vascularised by the CAM blood vessels and remained viable following implantation throughout the culture period. Histological evaluation showed formation of cell condensations in proximity to avian capillaries (immunocytochemistry positive for Sox9 and Runx2). New extracellular matrix (Collagen type II, X) deposition and osteoclast activity (Cathepsin K) on the CAM-implanted cylinders, correlated with a significant increase in bone volume as determined by µCT analysis (p<0.05) and demonstrated the ability of the human-avian system asa test/screen for biomaterials.
In summary, the current studies demonstrate the potential of the CAM assay as a surrogate blood supply for viable human bone tissue. This human-avian system offers a simple refinement model for animal research and a step towards a humanized in vivo model not only for tissue engineering, but for many other disciplines.

Text
full thesis v3 - Version of Record
Restricted to Repository staff only until 30 November 2021.
Available under License University of Southampton Thesis Licence.

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Published date: September 2016

Identifiers

Local EPrints ID: 436811
URI: http://eprints.soton.ac.uk/id/eprint/436811
PURE UUID: 582e63bf-82c6-4076-bf3f-412f98fba3a1
ORCID for Richard Oreffo: ORCID iD orcid.org/0000-0001-5995-6726

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Date deposited: 10 Jan 2020 17:31
Last modified: 18 Feb 2021 16:54

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