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Genetic manipulation of human mesenchymal progenitors to promote chondrogenesis using “bead-in-bead” polysaccharide capsules

Genetic manipulation of human mesenchymal progenitors to promote chondrogenesis using “bead-in-bead” polysaccharide capsules
Genetic manipulation of human mesenchymal progenitors to promote chondrogenesis using “bead-in-bead” polysaccharide capsules
Articular cartilage defects arising from trauma or degenerative diseases fail to repair spontaneously. We have adopted a non-viral gene delivery and tissue engineering strategy, in which Sox-9 transfected human mesenchymal progenitors have been encapsulated within alginate/chitosan polysaccharide capsules to promote chondrogenesis. Human bone marrow stromal cells and articular chondrocytes were transfected with flag-tagged Sox-9 plasmid and after 7 days in static culture, large regions of cell-generated matrix containing cartilage proteoglycans were observed as confirmed by positive Alcian blue staining and Sox-9 immunohistochemistry. Further, after 28 days, in vitro and in vivo, samples encapsulated with Sox-9 transfected cells demonstrated large regions of cartilaginous matrix as confirmed by positive Alcian blue staining, Sox-9 and type-II collagen immunohistochemistry, absent in samples encapsulated with untransfected cells. Extracted protein from in vivo constructs was further assessed by western blot analysis and positive expression of Sox-9 and type-II collagen was observed in Sox-9 transfected constructs which was absent in untransfected cells. Regions of cartilage-like matrix were significantly increased in Sox-9 constructs in comparison with untransfected constructs, confirming Sox-9 gene delivery enhances chondrogenesis in targeted cell populations, outlining the potential to promote cartilaginous construct formation with therapeutic implications for regeneration of human articular cartilage tissue defects.
chondrogenesis, human, cartilage, alginate, polysaccharide, gene transfer, gene expression
0142-9612
58-65
Babister, Jodie C.
3b35e463-b623-403c-91c3-88a590cea674
Tare, Rahul S.
587c9db4-e409-4e7c-a02a-677547ab724a
Green, David W.
9591e846-aeac-4549-b389-1744c5a43cff
Inglis, Stefanie
cb950bb9-8b89-41c0-b547-3206580d16b4
Mann, Stephen
1cbb2d4f-aee2-403c-9950-18bd388f02c0
Oreffo, RRichard O.C.
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Babister, Jodie C.
3b35e463-b623-403c-91c3-88a590cea674
Tare, Rahul S.
587c9db4-e409-4e7c-a02a-677547ab724a
Green, David W.
9591e846-aeac-4549-b389-1744c5a43cff
Inglis, Stefanie
cb950bb9-8b89-41c0-b547-3206580d16b4
Mann, Stephen
1cbb2d4f-aee2-403c-9950-18bd388f02c0
Oreffo, RRichard O.C.
ff9fff72-6855-4d0f-bfb2-311d0e8f3778

Babister, Jodie C., Tare, Rahul S., Green, David W., Inglis, Stefanie, Mann, Stephen and Oreffo, RRichard O.C. (2008) Genetic manipulation of human mesenchymal progenitors to promote chondrogenesis using “bead-in-bead” polysaccharide capsules. Biomaterials, 29 (1), 58-65. (doi:10.1016/j.biomaterials.2007.09.006). (PMID:17897711)

Record type: Article

Abstract

Articular cartilage defects arising from trauma or degenerative diseases fail to repair spontaneously. We have adopted a non-viral gene delivery and tissue engineering strategy, in which Sox-9 transfected human mesenchymal progenitors have been encapsulated within alginate/chitosan polysaccharide capsules to promote chondrogenesis. Human bone marrow stromal cells and articular chondrocytes were transfected with flag-tagged Sox-9 plasmid and after 7 days in static culture, large regions of cell-generated matrix containing cartilage proteoglycans were observed as confirmed by positive Alcian blue staining and Sox-9 immunohistochemistry. Further, after 28 days, in vitro and in vivo, samples encapsulated with Sox-9 transfected cells demonstrated large regions of cartilaginous matrix as confirmed by positive Alcian blue staining, Sox-9 and type-II collagen immunohistochemistry, absent in samples encapsulated with untransfected cells. Extracted protein from in vivo constructs was further assessed by western blot analysis and positive expression of Sox-9 and type-II collagen was observed in Sox-9 transfected constructs which was absent in untransfected cells. Regions of cartilage-like matrix were significantly increased in Sox-9 constructs in comparison with untransfected constructs, confirming Sox-9 gene delivery enhances chondrogenesis in targeted cell populations, outlining the potential to promote cartilaginous construct formation with therapeutic implications for regeneration of human articular cartilage tissue defects.

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More information

e-pub ahead of print date: 25 September 2007
Published date: January 2008
Keywords: chondrogenesis, human, cartilage, alginate, polysaccharide, gene transfer, gene expression

Identifiers

Local EPrints ID: 60880
URI: https://eprints.soton.ac.uk/id/eprint/60880
ISSN: 0142-9612
PURE UUID: b0889de8-5373-479f-834d-9225528cdb97
ORCID for Rahul S. Tare: ORCID iD orcid.org/0000-0001-8274-8837
ORCID for RRichard O.C. Oreffo: ORCID iD orcid.org/0000-0001-5995-6726

Catalogue record

Date deposited: 24 Sep 2008
Last modified: 19 Nov 2019 01:52

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