title: Nanoscale architecture and cellular adhesion of biomimetic collagen substrates.
abstract: The ability to engineer bioactive sites within the biopolymer collagen has significant potential to dictate cellular microenvironments and processes. We have developed a novel recombinant DNA platform that enables such molecular-level control over this important material. In this investigation, we demonstrated the production of synthetic human collagen using yeast strains that were engineered with human prolyl hydroxylase α and β genes integrated into the genome and a codon-optimized collagen gene carried on a plasmid. To understand the extent to which this synthetic collagen can mimic native human collagen, we examined the relationships between the structural topology and physical stability with the ability to support adhesion of HT-1080 cells. Characterization of these biopolymers included evaluation using circular dichroism spectroscopy, atomic force microscopy, and MTT metabolic activity assays. Although the apparent melting temperatures of the recombinant collagens were ∼3-5 less than native sources, the recombinant and native collagens exhibited comparable triple helical structure, polymeric dimensions, adsorption on polystyrene, and cellular adhesion properties below their respective melting temperature values. These results support the feasibility of producing molecularly-engineered collagens that can mimic native substrates for therapeutic and tissue engineering applications.
authors.affiliation: Department of Chemical Engineering and Materials Science, University of California, Irvine, CA, USA.
authors.forename: Abeer
authors.initials: A
authors.lastname: Jabaiah
authors.forename: Xi
authors.initials: X
authors.lastname: Wang
authors.forename: Senthil Kumar
authors.initials: SK
authors.lastname: Raman
authors.forename: Regina
authors.initials: R
authors.lastname: Ragan
authors.forename: Nancy A
authors.initials: NA
authors.lastname: Da Silva
authors.forename: Szu-Wen
authors.initials: SW
authors.lastname: Wang
mesh_terms: D000327:Adsorption; D032701:Biomimetics; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002942:Circular Dichroism; D003094:Collagen / Q000378:metabolism*; D006801:Humans; D018625:Microscopy, Atomic Force; D036103:Nanotechnology
chemical_list: D003094:Collagen
keywords: Collagen; atomic force microscopy; biomaterial; biomimetic; cell adhesion; recombinant protein; stability