Sulfated hyaluronan alters fibronectin matrix assembly and promotes osteogenic differentiation of human bone marrow stromal cells.
Extracellular matrix (ECM) composition and structural integrity is one of many factors that influence cellular differentiation. Fibronectin (FN) which is in many tissues the most abundant ECM protein forms a unique fibrillary network. FN homes several binding sites for sulfated glycosaminoglycans (sGAG), such as heparin (Hep), which was previously shown to influence FN conformation and protein binding. Synthetically sulfated hyaluronan derivatives (sHA) can serve as model molecules with a well characterized sulfation pattern to study sGAG-FN interaction. Here is shown that the low-sulfated sHA (sHA1) interacts with FN and influences fibril assembly. The interaction of FN fibrils with sHA1 and Hep, but not with non-sulfated HA was visualized by immunofluorescent co-staining. FRET analysis of FN confirmed the presence of more extended fibrils in human bone marrow stromal cells (hBMSC)-derived ECM in response to sHA1 and Hep. Although both sHA1 and Hep affected FN conformation, exclusively sHA1 increased FN protein level and led to thinner fibrils. Further, only sHA1 had a pro-osteogenic effect and enhanced the activity of tissue non-specific alkaline phosphatase. We hypothesize that the sHA1-triggered change in FN assembly influences the entire ECM network and could be the underlying mechanism for the pro-osteogenic effect of sHA1 on hBMSC.
Institute of Physiological Chemistry, Carl Gustav Carus Faculty of Medicine, Technische Universität Dresden, Fiedlerstrasse 42, 01307 Dresden, Germany.
Sarah
S
Vogel
Laboratory of Applied Mechanobiology, Department of Health Science and Technology, ETH Zurich, Vladimir-Prelog-Weg 4, 8093 Zurich, Switzerland.
Simon
S
Arnoldini
Biomaterials Department, INNOVENT e. V., Pruessingstrasse 27B, 07745 Jena, Germany.
Stephanie
S
Möller
Biomaterials Department, INNOVENT e. V., Pruessingstrasse 27B, 07745 Jena, Germany.
Matthias
M
Schnabelrauch
Institute of Physiological Chemistry, Carl Gustav Carus Faculty of Medicine, Technische Universität Dresden, Fiedlerstrasse 42, 01307 Dresden, Germany.
Ute
U
Hempel
D000469:Alkaline Phosphatase / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000378:metabolism; D005353:Fibronectins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D006025:Glycosaminoglycans / Q000737:chemistry / Q000378:metabolism; D006493:Heparin / Q000378:metabolism; D006801:Humans; D006820:Hyaluronic Acid / Q000378:metabolism / Q000494:pharmacology*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010012:Osteogenesis / Q000187:drug effects*; D011485:Protein Binding; D011487:Protein Conformation; D013431:Sulfates / Q000737:chemistry
D005353:Fibronectins; D006025:Glycosaminoglycans; D013431:Sulfates; D006820:Hyaluronic Acid; D006493:Heparin; D000469:Alkaline Phosphatase