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Bulk collagen incorporation rates into knitted stiff fibre polymer in tissue-engineered scaffolds: the rate-limiting step
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Materials Chemistry, Polymer Chemistry. (polymerkemi)
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Materials Chemistry, Polymer Chemistry. (polymerkemi)
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2008 (English)In: Journal of Tissue Engineering and Regenerative Medicine, ISSN 1932-6254, Vol. 2, no 8, 507-514 p.Article in journal (Refereed) Published
Abstract [en]

Fabrication of tissue-engineered constructs in vitro relies on sufficient synthesis of extracellular matrix (ECM) by cells to form a material suitable for normal function in vivo. Collagen synthesis by human dermal fibroblasts grown in vitro on two polymers, polyethylene terephthalate (PET) and polyglycolic acid (PGA), was measured by high-performance liquid chromatography (HPLC). Cells were either cultured in a dynamic environment, where meshes were loaded onto a pulsing tube in a bioreactor, or in a static environment without pulsing. Collagen synthesis by cells cultured on a static mesh increased by six-fold compared to monolayer culture, and increased by up to a further 5.4-fold in a pulsed bioreactor. However, little of the collagen synthesized was deposited onto the meshes, almost all being lost to the medium. The amount of collagen deposited onto meshes was highest when cells were cultured dynamically on PET meshes (17.6 µg), but deposition still represented only 1.4% of the total synthesized. Although total collagen synthesis was increased by the use of 3D culture and the introduction of pulsing, the results suggest that the limiting factor for fabrication of a tissue-engineered construct within practical timescales is not the amount of collagen synthesized but the quantity retained (i.e. deposited) within the construct during culture. This may be enhanced by systems which promote or assemble true 3D multi-layers of cells.

Place, publisher, year, edition, pages
John Wiley & Sons, Ltd. , 2008. Vol. 2, no 8, 507-514 p.
Keyword [en]
collagen synthesis, collagen deposition, tissue engineering, polyethylene terephthalate, 3D culture, bioreactors
National Category
Chemical Sciences
Research subject
Inorganic Chemistry
Identifiers
URN: urn:nbn:se:uu:diva-99347DOI: 10.1002/term.126ISI: 000262272900007OAI: oai:DiVA.org:uu-99347DiVA: diva2:207691
Available from: 2009-03-12 Created: 2009-03-12 Last updated: 2010-08-04Bibliographically approved
In thesis
1. Extracellular Matrix Based Materials for Tissue Engineering
Open this publication in new window or tab >>Extracellular Matrix Based Materials for Tissue Engineering
2010 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The extracellular matrix is (ECM) is a network of large, structural proteins and polysaccharides, important for cellular behavior, tissue development and maintenance. Present thesis describes work exploring ECM as scaffolds for tissue engineering by manipulating cells cultured in vitro or by influencing ECM expression in vivo. By culturing cells on polymer meshes under dynamic culture conditions, deposition of a complex ECM could be achieved, but with low yields. Since the major part of synthesized ECM diffused into the medium the rate limiting step of deposition was investigated. This quantitative analysis showed that the real rate limiting factor is the low proportion of new proteins which are deposited as functional ECM. It is suggested that cells are pre-embedded in for example collagen gels to increase the steric retention and hence functional deposition.

The possibility to induce endogenous ECM formation and tissue regeneration by implantation of growth factors in a carrier material was investigated. Bone morphogenetic protein-2 (BMP-2) is a growth factor known to be involved in growth and differentiation of bone and cartilage tissue. The BMP-2 processing and secretion was examined in two cell systems representing endochondral (chondrocytes) and intramembranous (mesenchymal stem cells) bone formation. It was discovered that chondrocytes are more efficient in producing BMP-2 compared to MSC. The role of the antagonist noggin was also investigated and was found to affect the stability of BMP-2 and modulate its effect. Finally, an injectable gel of the ECM component hyaluronan has been evaluated as delivery vehicle in cartilage regeneration. The hyaluronan hydrogel system showed promising results as a versatile biomaterial for cartilage regeneration, could easily be placed intraarticulary and can be used for both cell based and cell free therapies.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2010. 58 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 694
Keyword
Extracellular matrix, collagen synthesis, bioreactor, cell culture, bone morphogenetic protein-2, noggin, hyaluronan, cartilage
National Category
Biomedical Laboratory Science/Technology Cell and Molecular Biology Surgery Surgery Biomaterials Science Cell and Molecular Biology Biochemistry and Molecular Biology Polymer Chemistry Materials Chemistry
Research subject
Cell Research; Materials Science; Medical Cell Biology; Orthopaedics
Identifiers
urn:nbn:se:uu:diva-110631 (URN)978-91-554-7669-4 (ISBN)
Public defence
2010-01-15, Häggsalen, Ångströmlaboratoriet, Lägerhyddsvägen 1, Uppsala, 09:00 (English)
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Supervisors
Available from: 2009-12-18 Created: 2009-11-19 Last updated: 2009-12-18Bibliographically approved

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