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Publication : Thrombospondin-2 facilitates assembly of a type-I collagen-rich matrix in marrow stromal cells undergoing osteoblastic differentiation.

First Author  Alford AI Year  2013
Journal  Connect Tissue Res Volume  54
Issue  4-5 Pages  275-82
PubMed ID  23763373 Mgi Jnum  J:345745
Mgi Id  MGI:6852234 Doi  10.3109/03008207.2013.811236
Citation  Alford AI, et al. (2013) Thrombospondin-2 facilitates assembly of a type-I collagen-rich matrix in marrow stromal cells undergoing osteoblastic differentiation. Connect Tissue Res 54(4-5):275-82
abstractText  We examined the effects of Thrombospondin-2 (TSP2) deficiency on assembly of collagenous extracellular matrix (ECM) by primary marrow-derived mesenchymal stromal cells (MSC) undergoing osteoblast differentiation in culture. After 30 d, wild-type cells had accumulated and mineralized a collagen-rich insoluble matrix, whereas the TSP2-null cultures contained markedly lower amounts of matrix collagen and displayed reduced mineral. Differences in matrix collagen were seen as early as day 9, at which time wild-type cultures contained more total collagen per cell than did TSP2-null cells. Collagen was unevenly distributed amongst different extracellular compartments in the two cell-types. Collagen levels in conditioned medium of wild-type cells were higher than those of TSP2-null cells, but were roughly equivalent in the acid-soluble, newly cross-linked matrixes. Conversely, the mature, cross-linked acid-insoluble matrix layer of wild-type cells contained about twice as much collagen as TSP2-null cell-derived matrix. Western blot analysis of type-I collagen in detergent-soluble and insoluble matrix fractions supported the premise that matrix collagen levels were reduced in TSP2-null MSC undergoing osteoblastic differentiation in vitro. Western blot and immunofluorescent analysis suggested that assembly of fibronectin into matrix was not affected by TSP2 deficiency. Instead, western blots of conditioned medium demonstrated a marked reduction in mature, fully processed type-I collagen in the absence of TSP2. Our data suggest that in the context of osteoblast differentiation, TSP2 promotes the assembly of a type-I collagen-rich matrix by facilitating pro-collagen processing.
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