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dc.contributor.authorMagnus Østgård, Olderøy
dc.contributor.authorLilledahl, Magnus Borstad
dc.contributor.authorBeckwith, Marianne
dc.contributor.authorMelvik, Jan Egil
dc.contributor.authorReinholt, Finn P.
dc.contributor.authorSikorski, Pawel
dc.contributor.authorBrinchmann, Jan E.
dc.date.accessioned2015-11-24T12:39:38Z
dc.date.accessioned2015-12-09T12:48:06Z
dc.date.available2015-11-24T12:39:38Z
dc.date.available2015-12-09T12:48:06Z
dc.date.issued2014
dc.identifier.citationPLoS ONE 2014, 9(3):e91662nb_NO
dc.identifier.issn1932-6203
dc.identifier.urihttp://hdl.handle.net/11250/2367367
dc.description.abstractA popular approach to make neocartilage in vitro is to immobilize cells with chondrogenic potential in hydrogels. However, functional cartilage cannot be obtained by control of cells only, as function of cartilage is largely dictated by architecture of extracellular matrix (ECM). Therefore, characterization of the cells, coupled with structural and biochemical characterization of ECM, is essential in understanding neocartilage assembly to create functional implants in vitro. We focused on mesenchymal stem cells (MSC) immobilized in alginate hydrogels, and used immunohistochemistry (IHC) and gene expression analysis combined with advanced microscopy techniques to describe properties of cells and distribution and organization of the forming ECM. In particular, we used second harmonic generation (SHG) microscopy and focused ion beam/scanning electron microscopy (FIB/SEM) to study distribution and assembly of collagen. Samples with low cell seeding density (1e7 MSC/ml) showed type II collagen molecules distributed evenly through the hydrogel. However, SHG microscopy clearly indicated only pericellular localization of assembled fibrils. Their distribution was improved in hydrogels seeded with 5e7 MSC/ml. In those samples, FIB/SEM with nm resolution was used to visualize distribution of collagen fibrils in a three dimensional network extending from the pericellular region into the ECM. In addition, distribution of enzymes involved in procollagen processing were investigated in the alginate hydrogel by IHC. It was discovered that, at high cell seeding density, procollagen processing and fibril assembly was also occurring far away from the cell surface, indicating sufficient transport of procollagen and enzymes in the intercellular space. At lower cell seeding density, the concentration of enzymes involved in procollagen processing was presumably too low. FIB/SEM and SHG microscopy combined with IHC localization of specific proteins were shown to provide meaningful insight into ECM assembly of neocartilage, which will lead to better understanding of cartilage formation and development of new tissue engineering strategies.nb_NO
dc.language.isoengnb_NO
dc.publisherPublic Library of Sciencenb_NO
dc.titleBiochemical and Structural Characterization of Neocartilage Formed by Mesenchymal Stem Cells in Alginate Hydrogelsnb_NO
dc.typeJournal articlenb_NO
dc.typePeer revieweden_GB
dc.date.updated2015-11-24T12:39:38Z
dc.source.volume9nb_NO
dc.source.journalPLoS ONEnb_NO
dc.source.issue3nb_NO
dc.identifier.doi10.1371/journal.pone.0091662
dc.identifier.cristin1152133
dc.description.localcode© 2014 Olderøy et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.nb_NO


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