The biological effects of fibrin-binding synthetic oligopeptides derived from fibronectin on osteoblast-like cells
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Purpose: The aim of this study was to investigate the effects of synthetic fibronectin (FN) fragments, including fibrin binding sites from amino-terminal FN fragments containing type I repeats 1 to 5, on osteoblast-like cell activity. Methods: Oligopeptides ranging from 9 to 20 amino acids, designated FF1, FF3, and FF5, were synthesized by a solid-phase peptide synthesizing system, and we investigated the effects of these peptides on cell attachment and extent of mineralization using confocal microscopy, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays, and Alizarin red S staining. Results: FF3 and FF5 peptides increased the number of attached human osteoblastic cells, and FF3 administration led to prominent cell spreading. Mineralization was increased in FF3 and FF5 compared to FF1 and the untreated control. Conclusions: Taken together, it can be concluded that the fibrin-binding oligopeptides FF3 and FF5 enhanced cell attachment and mineralization on osteoblast-like cells. These results indicate that FF3 and FF5 have the potential to increase osteoblast-like cell activity. Performing an in vivo study may provide further possibilities for surface modification of biomimetic peptides to enhance osteogenesis, thus improving the regeneration of destroyed alveolar bone.
Abstract
Purpose: The aim of this study was to investigate the effects of synthetic fibronectin (FN) fragments, including fibrin binding sites from amino-terminal FN fragments containing type I repeats 1 to 5, on osteoblast-like cell activity. Methods: Oligopeptides ranging from 9 to 20 amino acids, designated FF1, FF3, and FF5, were synthesized by a solid-phase peptide synthesizing system, and we investigated the effects of these peptides on cell attachment and extent of mineralization using confocal microscopy, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays, and Alizarin red S staining. Results: FF3 and FF5 peptides increased the number of attached human osteoblastic cells, and FF3 administration led to prominent cell spreading. Mineralization was increased in FF3 and FF5 compared to FF1 and the untreated control. Conclusions: Taken together, it can be concluded that the fibrin-binding oligopeptides FF3 and FF5 enhanced cell attachment and mineralization on osteoblast-like cells. These results indicate that FF3 and FF5 have the potential to increase osteoblast-like cell activity. Performing an in vivo study may provide further possibilities for surface modification of biomimetic peptides to enhance osteogenesis, thus improving the regeneration of destroyed alveolar bone.
참고문헌 (34)
- Damsky CH, "Extracellular matrix-integrin interactions in osteoblast function and tissue remodeling", Bone, 25, pp.95~96 (1999)
- Embery G, "Connective tissue elements as diagnostic aids in periodontology", Periodontol 2000, 24, pp.193~214 (2000)
- Rosso F, "From cell-ECM interactions to tissue engineering", J Cell Physiol, 199, pp.174~180 (2004)
- Mosher DF, "Fibronectin" (1989), Academic Press
- Garcia AJ, "Effect of surface reaction stage on fibronectin-mediated adhesion of osteoblast-like cells to bioactive glass", J Biomed Mater Res, 40, pp.48~56 (1998)
- Ding HT, "Fibronectin enhances in vitro vascular calcification by promoting osteoblastic differentiation of vascular smooth muscle cells via ERK pathway", J Cell Biochem, 99, pp.1343~1352 (2006)
- Majmudar G, "Bone cell culture in a three-dimensional polymer bead stabilizes the differentiated phenotype and provides evidence that osteoblastic cells synthesize type III collagen and fibronectin", J Bone Miner Res, 6, pp.869~881 (1991)
- Pankov R, "Fibronectin at a glance", J Cell Sci, 115, pp.3861~3863 (2002)
- Mardon HJ, "The role of the ninth and tenth type III domains of human fibronectin in cell adhesion", FEBS Lett, 340, pp.197~201 (1994)
- Redick SD, "Defining fibronectin’s cell adhesion synergy site by site-directed mutagenesis", J Cell Biol, 149, pp.521~527 (2000)
- Magnusson MK, "Fibronectin: structure, assembly, and cardiovascular implications", Arterioscler Thromb Vasc Biol, 18, pp.1363~1370 (1998)
- Corbett SA, "Covalent cross-linking of fibronectin to fibrin is required for maximal cell adhesion to a fibronectin-fibrin matrix", J Biol Chem, 272, pp.24999~25005 (1997)
- Grinnell F, "Fibroblast adhesion to fibrinogen and fibrin substrata: requirement for cold-insoluble globulin (plasma fibronectin)", Cell, 19, pp.517~525 (1980)
- Corbett SA, "Changes in cell spreading and cytoskeletal organization are induced by adhesion to a fibronectin-fibrin matrix", Blood, 88, pp.158~166 (1996)
- Greiling D, "Fibronectin provides a conduit for fibroblast transmigration from collagenous stroma into fibrin clot provisional matrix", J Cell Sci, 110, pp.861~870 (1997)
- Clark RA, "Fibroblast invasive migration into fibronectin", fibrin gels requires a previously uncharacterized dermatan sulfate-CD44 proteoglycan, 2004, pp.122, 266~277
- Knox P, "Role of fibronectin in the migration of fibroblasts into plasma clots", J Cell Biol, 102, pp.2318~2323 (1986)
- Rostagno AA, "Comparison of the fibrin-binding activities in the N- and C-termini of fibronectin", Biochem J, 338, pp.375~386 (1999)
- Matsuka YV, "The NH2-terminal fibrin-binding site of fibronectin is formed by interacting fourth and fifth finger domains. Studies with recombinant finger fragments expressed in Escherichia coli", J Biol Chem, 269, pp.9539~9546 (1994)
- Garcia-Pardo A, "Primary structure of human plasma fibronectin. Characterization of a 31,000-dalton fragment from the COOH-terminal region containing a free sulfhydryl group and a fibrin-binding site", J Biol Chem, 260, pp.10320~10325 (1985)
- Cutler SM, "Engineering cell adhesive surfaces that direct integrin alpha5beta1 binding using a recombinant fragment of fibronectin", Biomaterials, 24, pp.1759~1770 (2003)
- Kim TI, "Fibronectin fragment promotes osteoblast-associated gene expression and biological activity of human osteoblast-like cell", Biotechnol Lett, 25, pp.2007~2011 (2003)
- Kim TI, "Biomimetic approach on human periodontal ligament cells using synthetic oligopeptides", J Periodontol, 75, pp.925~932 (2004)
- Ku Y, "The effect of the surface modification of titanium using a recombinant fragment of fibronectin and vitronectin on cell behavior", Biomaterials, 26, pp.5153~5157 (2005)
- Jaiswal N, "Osteogenic differentiation of purified, culture-expanded human mesenchymal stem cells in vitro", J Cell Biochem, 64, pp.295~312 (1997)
- Koch TG, "Isolation of mesenchymal stem cells from equine umbilical cord blood", BMC Biotechnol, 7, pp.26~ (2007)
- Clover J, "Are MG-63 and HOS TE85 human osteosarcoma cell lines representative models of the osteoblastic phenotype?", Bone, 15, pp.585~591 (1994)
- Clark RA, "Fibroblast migration on fibronectin requires three distinct functional domains", J Invest Dermatol, 121, pp.695~705 (2003)
- Purohit A, "Estrogen synthesis by osteoblast cell lines", Endocrinology, 131, pp.2027~2029 (1992)
- Kinpara K, "Osteoclast differentiation factor in human osteosarcoma cell line", J Immunoassay, 21, pp.327~340 (2000)
- Rostagno A, "Further characterization of the NH2-terminal fibrin-binding site on fibronectin", J Biol Chem, 269, pp.31938~31945 (1994)
- Williams MJ, "Solution structure of a pair of fibronectin type 1 modules with fibrin binding activity", J Mol Biol, 235, pp.1302~1311 (1994)
- Blystone SD, "Fibronectin dependent macrophage fibrin binding", Blood, 78, pp.2900~2907 (1991)
- Magnusson MK, "Fibronectin: structure, assembly, and cardiovascular implications", Arterioscler Thromb Vasc Biol, 18, pp.1363~1370 (1998)
