title: Covalent binding of BMP-2 on surfaces using a self-assembled monolayer approach.
abstract: Bone morphogenetic protein 2 (BMP-2) is a growth factor embedded in the extracellular matrix of bone tissue. BMP-2 acts as trigger of mesenchymal cell differentiation into osteoblasts, thus stimulating healing and de novo bone formation. The clinical use of recombinant human BMP-2 (rhBMP-2) in conjunction with scaffolds has raised recent controversies, based on the mode of presentation and the amount to be delivered. The protocol presented here provides a simple and efficient way to deliver BMP-2 for in vitro studies on cells. We describe how to form a self-assembled monolayer consisting of a heterobifunctional linker, and show the subsequent binding step to obtain covalent immobilization of rhBMP-2. With this approach it is possible to achieve a sustained presentation of BMP-2 while maintaining the biological activity of the protein. In fact, the surface immobilization of BMP-2 allows targeted investigations by preventing unspecific adsorption, while reducing the amount of growth factor and, most notably, hindering uncontrolled release from the surface. Both short- and long-term signaling events triggered by BMP-2 are taking place when cells are exposed to surfaces presenting covalently immobilized rhBMP-2, making this approach suitable for in vitro studies on cell responses to BMP-2 stimulation.
authors.affiliation: Department of Biophysical Chemistry, Institute for Physical Chemistry, University of Heidelberg.
authors.forename: Theresa L M
authors.initials: TL
authors.lastname: Pohl
authors.forename: Elisabeth H
authors.initials: EH
authors.lastname: Schwab
authors.forename: Elisabetta A
authors.initials: EA
authors.lastname: Cavalcanti-Adam
mesh_terms: D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000737:chemistry* / Q000378:metabolism; D002460:Cell Line; D006046:Gold / Q000737:chemistry; D055767:Immobilized Proteins / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D032446:Myoblasts
chemical_list: D055396:Bone Morphogenetic Protein 2; D055767:Immobilized Proteins; D006046:Gold
references.citation: Biosens Bioelectron. 2006 Aug 15;22(2):233-40
references.pmid: 16460923
references.citation: Biomaterials. 2009 Feb;30(6):1166-75
references.pmid: 19022501
references.citation: Genes Dev. 1996 Jul 1;10(13):1580-94
references.pmid: 8682290
references.citation: J Biomed Mater Res A. 2004 Dec 1;71(3):528-37
references.pmid: 15478212
references.citation: Bioconjug Chem. 2010 Oct 20;21(10):1762-72
references.pmid: 20886828
references.citation: Chem Rev. 2005 Apr;105(4):1103-69
references.pmid: 15826011
references.citation: Acta Biomater. 2012 Feb;8(2):772-80
references.pmid: 22040684
references.citation: Biomaterials. 2002 May;23(9):2079-87
references.pmid: 11996050
references.citation: J Pharm Pharmacol. 2001 Nov;53(11):1427-37
references.pmid: 11732745
references.citation: J Cell Biol. 1994 Dec;127(6 Pt 1):1755-66
references.pmid: 7798324
references.citation: Eur J Pharm Biopharm. 2004 Sep;58(2):197-208
references.pmid: 15296949
references.citation: Nat Struct Mol Biol. 2004 May;11(5):481-8
references.pmid: 15064755
references.citation: Neurosurg Focus. 2002 Dec 15;13(6):e2
references.pmid: 15766228
references.citation: Biomaterials. 2007 Aug;28(24):3517-29
references.pmid: 17482256
references.citation: J Biomed Mater Res. 2000 May;50(2):227-38
references.pmid: 10679688
references.citation: Ann N Y Acad Sci. 2006 Apr;1068:19-25
references.pmid: 16831902
references.citation: Biomaterials. 1997 Sep;18(18):1201-25
references.pmid: 9300556
references.citation: Macromol Biosci. 2011 Sep 9;11(9):1149-63
references.pmid: 21509937
references.citation: Langmuir. 2005 Aug 2;21(16):7071-5
references.pmid: 16042424
references.citation: Eur J Biochem. 1996 Apr 1;237(1):295-302
references.pmid: 8620887
references.citation: J Mol Biol. 1999 Mar 19;287(1):103-15
references.pmid: 10074410
references.citation: Adv Mater. 2011 Mar 25;23(12):H111-8
references.pmid: 21433098
references.citation: Cytokine Growth Factor Rev. 2005 Jun;16(3):249-50
references.pmid: 15949967
references.citation: Cytokine Growth Factor Rev. 2009 Oct-Dec;20(5-6):343-55
references.pmid: 19897402
references.citation: Nature. 1992 Jul 30;358(6385):430-4
references.pmid: 1641027
references.citation: Cytokine Growth Factor Rev. 2005 Jun;16(3):251-63
references.pmid: 15871923
references.citation: Clin Orthop Relat Res. 1998 Jan;(346):26-37
references.pmid: 9577407
references.citation: J Pharm Pharmacol. 2004 Apr;56(4):415-27
references.pmid: 15099436
references.citation: Spine J. 2011 Jun;11(6):471-91
references.pmid: 21729796
references.citation: J Biol Chem. 2002 Feb 15;277(7):5330-8
references.pmid: 11714695
references.citation: J Biomed Mater Res A. 2010 Aug;94(2):576-85
references.pmid: 20198690
references.citation: Nat Struct Biol. 2000 Jun;7(6):492-6
references.pmid: 10881198
references.citation: Biotechnol Bioeng. 2007 Dec 15;98(6):1109-22
references.pmid: 17514756
references.citation: Biomaterials. 2003 Nov;24(24):4385-415
references.pmid: 12922151
references.citation: J Biomed Mater Res A. 2009 Jan;88(1):162-73
references.pmid: 18286636