title: Recombinant human type II collagen hydrogel provides a xeno-free 3D micro-environment for chondrogenesis of human bone marrow-derived mesenchymal stromal cells.
abstract: Recombinant human type II collagen (rhCII) hydrogel was tested as a xeno-free micro-environment for the chondrogenesis of human bone marrow-derived mesenchymal stromal cells (BM-MSCs). The rhCII hydrogels were seeded with BM-MSCs and cultured in a xeno-free chondro-inductive medium for 14, 28 and 84 days. High-density pellet cultures served as controls. The samples were subjected to biochemical, histological and gene expression analyses. Although the cells deposited glycosaminoglycans into the extracellular space significantly more slowly in the rhCII hydrogels compared to the high-density pellets, a similar potential of matrix deposition was reached by the end of the 84-day culture. At day 28 of culture, the gene expression level for cartilage marker genes (i.e. genes encoding for Sox9 transcription factor, Collagen type II and Aggrecan) were considerably lower in the rhCII hydrogels than in the high-density pellets, but at the end of the 84-day culture period, all the cartilage marker genes analysed were expressed at a similar level. Interestingly, the expression of the matrix metallopeptidases (MMP)-13, MMP-14 and MMP-8, i.e. extracellular collagen network-degrading enzymes, were transiently upregulated in the rhCII hydrogel, indicating active matrix reorganization. This study demonstrated that the rhCII hydrogel functions as a xeno-free platform for BM-MSC chondrogenesis, although the process is delayed. The reversible catabolic reaction evoked by the rhCII hydrogel might be beneficial in graft integration in vivo and pinpoints the need to further explore the use of hydrogels containing recombinant extracellular matrix (ECM) proteins to induce the chondrogenesis of MSCs. Copyright © 2015 John Wiley & Sons, Ltd.
authors: Department of Surgery, Institute of Clinical Medicine, University of Helsinki, Finland.|Department of Orthopaedics and Traumatology, Hospital District of Helsinki and Uusimaa, Helsinki, Finland.
authors: Virpi
authors: V
authors: Muhonen
authors: Department of Orthopaedics, Erasmus MC, University Medical Centre, Rotterdam, The Netherlands.
authors: Roberto
authors: R
authors: Narcisi
authors: Finnish Red Cross Blood Service, Research and Cell Therapy Services, Helsinki, Finland.
authors: Johanna
authors: J
authors: Nystedt
authors: Finnish Red Cross Blood Service, Research and Cell Therapy Services, Helsinki, Finland.
authors: Matti
authors: M
authors: Korhonen
authors: Department of Orthopaedics, Erasmus MC, University Medical Centre, Rotterdam, The Netherlands.|Department of Otorhinolaryngology, Erasmus MC, University Medical Centre, Rotterdam, The Netherlands.
authors: Gerjo J V M
authors: GJ
authors: van Osch
authors: Department of Surgery, Institute of Clinical Medicine, University of Helsinki, Finland.|Department of Orthopaedics and Traumatology, Hospital District of Helsinki and Uusimaa, Helsinki, Finland.
authors: Ilkka
authors: I
authors: Kiviranta
mesh_terms: D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D002356:Cartilage; D060833:Cellular Microenvironment / Q000187:drug effects*; D020219:Chondrogenesis / Q000187:drug effects*; D024043:Collagen Type II / Q000494:pharmacology*; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000494:pharmacology*; D007150:Immunohistochemistry; D020782:Matrix Metalloproteinases / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology*
chemical_list: D024043:Collagen Type II; D006025:Glycosaminoglycans; D011994:Recombinant Proteins; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D012313:RNA; D020782:Matrix Metalloproteinases
keywords: cartilage repair; chondrogenesis; matrix metallopeptidase; mesenchymal stem cells; recombinant collagen hydrogel; xeno-free