Cell behavior on a CCN1 functionalized elastin-mimetic protein polymer.
We report the design of an elastin-mimetic triblock copolymer with the ability to guide endothelial cell adhesion, spreading, and migration while maintaining the elastomeric properties of the protein polymer. The V2 ligand sequence from matricellular protein CCN1 (cysteine-rich 61, CYR61) was multimerized and cloned into elastin polymer LysB10, creating LysB10.V2. Cell adhesion studies demonstrated that a LysB10.V2 surface density of at least 40 pmol/cm(2) was required to elicit cell attachment. Peptide blocking studies confirmed V2 specific engagement with integrin receptor α(v)β(3) (P < 0.05) and we observed the formation of actin stress fiber networks and vinculin clustering, characteristic of focal adhesion assembly. Haptotatic migration assays demonstrated the ability of LysB10.V2 surfaces to stimulate migration of endothelial cells (P < 0.05). Significantly, we illustrated the ability of LysB10.V2 to support a quiescent endothelium. The CCN1 molecule functions to support many key biological processes necessary for tissue repair and thus presents a promising target for bioengineering applications. Collectively, our results demonstrate the potential to harness CCN1 specific function in the design of new scaffold materials for applications in regenerative medicine.
Department of Biomedical Engineering, Emory University/Georgia Institute of Technology, Atlanta, GA 30332, United States.
Swathi
S
Ravi
Carolyn A
CA
Haller
Rory E
RE
Sallach
Elliot L
EL
Chaikof
D000595:Amino Acid Sequence; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D003432:Cross-Linking Reagents / Q000494:pharmacology; D055515:Cysteine-Rich Protein 61 / Q000494:pharmacology*; D004549:Elastin / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D022001:Focal Adhesions / Q000187:drug effects / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D020100:Hydrogels / Q000494:pharmacology; D016023:Integrins / Q000378:metabolism; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary
D003432:Cross-Linking Reagents; D055515:Cysteine-Rich Protein 61; D020100:Hydrogels; D016023:Integrins; D004549:Elastin