title: Photoinitiator-free synthesis of endothelial cell-adhesive and enzymatically degradable hydrogels.
abstract: We report on a photoinitiator-free synthetic method of incorporating bioactivity into poly(ethylene glycol) (PEG) hydrogels in order to control physical properties, enzymatic biodegradability and cell-specific adhesiveness of the polymer network, while eliminating the need for UV-mediated photopolymerization. To accomplish this, hydrogel networks were polymerized using Michael addition with four-arm PEG acrylate (10 kDa), using a collagenase-sensitive peptide (CSP) as a crosslinker, and introducing an endothelial cell-adhesive peptide either terminally (RGD) or attached to the crosslinking peptide sequence (CSP-RGD). The efficiency of the Michael addition reactions were determined by nuclear magnetic resonance and Ellman's assay. Successful decoupling of cell adhesivity and physical properties was demonstrated by quantifying and comparing the swelling ratios and Young's moduli of various hydrogel formulations. Degradation profiles were established by incubating functionalized hydrogels in collagenase solutions (0.0-1.0 μg ml(-1)), demonstrating that functionalized hydrogels degraded at a rate dependent upon collagenase concentration. Moreover, it was shown that the degradation rate was independent of CSP-RGD concentration. Cell attachment and proliferation on functionalized hydrogels were compared for various RGD concentrations, providing evidence that cell attachment and proliferation were directly related to relative amounts of the CSP-RGD combination peptide. An increase in cell viability was achieved using Michael addition techniques when compared to UV polymerization, and was assessed by a LIVE/DEAD fluorescence assay. This photoinitiator-free method shows promise in creating hydrogel-based tissue engineering scaffolds allow for decoupled cell adhesivity and physical properties and that render greater cell viability.
authors.affiliation: Department of Biomedical Engineering, Case Western Reserve University, Cleveland, OH 44106, USA.
authors.forename: Derek R
authors.initials: DR
authors.lastname: Jones
authors.affiliation: Department of Biomedical Engineering, Case Western Reserve University, Cleveland, OH 44106, USA.
authors.forename: Roger E
authors.initials: RE
authors.lastname: Marchant
authors.affiliation: Department of Biomedical Engineering, Case Western Reserve University, Cleveland, OH 44106, USA.
authors.forename: Horst
authors.initials: H
authors.lastname: von Recum
authors.affiliation: Department of Biomedical Engineering, Case Western Reserve University, Cleveland, OH 44106, USA.
authors.forename: Anirban
authors.initials: A
authors.lastname: Sen Gupta
authors.affiliation: Department of Biomedical Engineering, Case Western Reserve University, Cleveland, OH 44106, USA; Robert J. Tomsich Pathology and Laboratory Medicine Institute, Cleveland Clinic, Cleveland, OH 44195, USA. Electronic address: marchak@ccf.org.
authors.forename: Kandice
authors.initials: K
authors.lastname: Kottke-Marchant
mesh_terms: D000179:Acrylates / Q000737:chemistry*; D063566:Biodegradable Plastics / Q000737:chemistry*; D002448:Cell Adhesion; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D020100:Hydrogels / Q000138:chemical synthesis / Q000737:chemistry; D009842:Oligopeptides / Q000737:chemistry; D055668:Photochemical Processes; D011092:Polyethylene Glycols / Q000737:chemistry*; D014466:Ultraviolet Rays
chemical_list: D000179:Acrylates; D063566:Biodegradable Plastics; D020100:Hydrogels; D009842:Oligopeptides; D011092:Polyethylene Glycols; C047981:arginyl-glycyl-aspartic acid; C036658:acrylic acid
keywords: Cell adhesive; Enzyme degradable; Extracellular matrix; Michael addition; Photoinitiator free