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Development of in vivo muCT evaluation of neovascularisation in tissue engineered bone constructs

Development of in vivo muCT evaluation of neovascularisation in tissue engineered bone constructs
Development of in vivo muCT evaluation of neovascularisation in tissue engineered bone constructs
Due to an increasing aging population the need for innovative approaches to aid skeletal repair and reconstruction is a significant socio-economic increasing problem. The emerging discipline of tissue engineering has sort to augment the growth and repair of bone loss particularly in areas of trauma, degeneration and revision surgery. However, the initiation and development of a fully functional vascular network are critical for bioengineered bone to repair large osseous defects, whether the material is osteosynthetic (poly (d,l)-lactic acid, PLA) or natural bone allograft. Quantification and three-dimensional visualization of new vessel networks remain a problem in bone tissue engineering constructs. A novel technique utilising a radio-opaque dye and micro-computed tomography (muCT) has been developed and applied to study angiogenesis in an impaction bone graft model. Tissue-engineered constructs combining human bone marrow stromal cells (HBMSC) with natural allograft and synthetic grafts (PLA) were impacted and implanted into the subcutis of MF-1 nu/nu mice for a period of 28 days. Microfil consisting of radio-opaque polymer was perfused through the mice and scanned using a Bench Top CT system for micro-computed tomography. Analysis of three-dimensional muCT reconstructions demonstrated an increase in vessel volume and vessel number in the impacted scaffolds/HBMC compared to scaffolds alone. Vessel volume: allograft/HBMSC=0.57 mm(3)+/-0.19; allograft=0.04 mm(3)+/-0.04; PLA/HBMSC=1.19 mm(3)+/-0.31; and PLA=0.12 mm(3)+/-0.01. Penetrating vessel number: allograft/HBMSC=22.33+/-3.21; allograft=3.67+/-1.153; PLA/HBMSC=32.67+/-8.33; and PLA=7.67+/-3.06. Type 1 collagen and von Willebrand factor immunohistochemistry in scaffold/HBMSC constructs indicated the osteogenic cell phenotype, and new blood vessel formation respectively. Contrast-enhanced 3D reconstructions facilitated the visualization and quantification of neovascularisation. This novel technique has been used to demonstrate neovascularisation in impacted tissue engineered constructs providing a facile approach with wide experimental application.
development, phenotype, growth, human, von willebrand factor, human development, bone marrow, blood, tissue engineering, surgery, research, mice, regeneration, aging, stem cells, immunohistochemistry, analysis, acid, bone, stromal cells, vascular
8756-3282
195-202
Bolland, B.J.
c8fd42ef-2524-4eaf-b1d4-10281cdedebb
Kanczler, J.M.
eb8db9ff-a038-475f-9030-48eef2b0559c
Dunlop, D.G.
5f8d8b5c-e516-48b8-831f-c6e5529a52cc
Oreffo, R.O.
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Bolland, B.J.
c8fd42ef-2524-4eaf-b1d4-10281cdedebb
Kanczler, J.M.
eb8db9ff-a038-475f-9030-48eef2b0559c
Dunlop, D.G.
5f8d8b5c-e516-48b8-831f-c6e5529a52cc
Oreffo, R.O.
ff9fff72-6855-4d0f-bfb2-311d0e8f3778

Bolland, B.J., Kanczler, J.M., Dunlop, D.G. and Oreffo, R.O. (2008) Development of in vivo muCT evaluation of neovascularisation in tissue engineered bone constructs. Bone, 43 (1), 195-202. (doi:10.1016/j.bone.2008.02.013).

Record type: Article

Abstract

Due to an increasing aging population the need for innovative approaches to aid skeletal repair and reconstruction is a significant socio-economic increasing problem. The emerging discipline of tissue engineering has sort to augment the growth and repair of bone loss particularly in areas of trauma, degeneration and revision surgery. However, the initiation and development of a fully functional vascular network are critical for bioengineered bone to repair large osseous defects, whether the material is osteosynthetic (poly (d,l)-lactic acid, PLA) or natural bone allograft. Quantification and three-dimensional visualization of new vessel networks remain a problem in bone tissue engineering constructs. A novel technique utilising a radio-opaque dye and micro-computed tomography (muCT) has been developed and applied to study angiogenesis in an impaction bone graft model. Tissue-engineered constructs combining human bone marrow stromal cells (HBMSC) with natural allograft and synthetic grafts (PLA) were impacted and implanted into the subcutis of MF-1 nu/nu mice for a period of 28 days. Microfil consisting of radio-opaque polymer was perfused through the mice and scanned using a Bench Top CT system for micro-computed tomography. Analysis of three-dimensional muCT reconstructions demonstrated an increase in vessel volume and vessel number in the impacted scaffolds/HBMC compared to scaffolds alone. Vessel volume: allograft/HBMSC=0.57 mm(3)+/-0.19; allograft=0.04 mm(3)+/-0.04; PLA/HBMSC=1.19 mm(3)+/-0.31; and PLA=0.12 mm(3)+/-0.01. Penetrating vessel number: allograft/HBMSC=22.33+/-3.21; allograft=3.67+/-1.153; PLA/HBMSC=32.67+/-8.33; and PLA=7.67+/-3.06. Type 1 collagen and von Willebrand factor immunohistochemistry in scaffold/HBMSC constructs indicated the osteogenic cell phenotype, and new blood vessel formation respectively. Contrast-enhanced 3D reconstructions facilitated the visualization and quantification of neovascularisation. This novel technique has been used to demonstrate neovascularisation in impacted tissue engineered constructs providing a facile approach with wide experimental application.

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More information

Published date: July 2008
Keywords: development, phenotype, growth, human, von willebrand factor, human development, bone marrow, blood, tissue engineering, surgery, research, mice, regeneration, aging, stem cells, immunohistochemistry, analysis, acid, bone, stromal cells, vascular

Identifiers

Local EPrints ID: 60924
URI: https://eprints.soton.ac.uk/id/eprint/60924
ISSN: 8756-3282
PURE UUID: a4b48d29-9cd0-4210-826f-d7356f688367
ORCID for J.M. Kanczler: ORCID iD orcid.org/0000-0001-7249-0414
ORCID for R.O. Oreffo: ORCID iD orcid.org/0000-0001-5995-6726

Catalogue record

Date deposited: 03 Sep 2008
Last modified: 14 Mar 2019 01:48

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Contributors

Author: B.J. Bolland
Author: J.M. Kanczler ORCID iD
Author: D.G. Dunlop
Author: R.O. Oreffo ORCID iD

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