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Title

QUANTITATIVE ANALYSIS OF THE EFFECTS OF BIOFUNCTIONAL AND PHYSICAL GRADIENTS ON CELL BEHAVIOR IN POLY (ETHYLENE GLYCOL) DIACRYLATE HYDROGELS

Creator

Turturro, Michael

Date

2012-10-29, 2012-12

Description

The continued enhancement of tissue engineered scaffolds relies on their ability to stimulate the formation of a stable microvascular network...
Show moreThe continued enhancement of tissue engineered scaffolds relies on their ability to stimulate the formation of a stable microvascular network within the biomaterial. In vivo, the spatial presentation of immobilized extracellular matrix cues and matrix mechanical properties play an important role in directed and guided cell behavior and neovascularization. The overall goals of this thesis are to develop a technique for the generation of gradients of physical properties and incorporated biofunctionality within poly(ethylene glycol) diacrylate (PEGDA) scaffolds and to investigate the effects of these gradients on 3D cell invasion and neovascularization. To this end, a novel photopolymerization technique for generating spatial variations in matrix properties and incorporated biofunctionality within synthetic PEGDA hydrogels, perfusion-based frontal polymerization (PBFP), was developed. This technique relies on the controlled perfusion of a photoinitiator to a reaction chamber containing a precursor solution and results in the propagation of a polymer reaction front that travels through the monomer solution creating a gradient in hydrogel crosslinking. Manipulation of the magnitude of the gradient can be achieved through alterations in the polymerization conditions. Scaffolds with embedded gradients were designed and optimized based on a range of properties shown to support 2D cell adhesion, proliferation, and 3D vascular cell invasion in bulk photopolymerized hydrogels with homogeneous properties. An in vitro model of neovascularization was used to evaluate the effect of these gradients on vascular sprout formation. Sprout invasion in gradient hydrogels occurred bi-directionally with sprout alignment observed in the direction parallel to the gradient while control hydrogels with homogeneous properties resulted in uniform invasion. In PBFP gradient hydrogels, sprout xvi length was found to be twice as long in the direction parallel to the gradient as compared to the perpendicular direction after three weeks in culture. This directionality was found to be more prominent in gradient regions of increased stiffness, crosslinked matrix metalloproteinase (MMP)-sensitive peptide presentation, and immobilized YRGDS concentration. In vivo tissue invasion was shown to be directly related to gradient properties and orientation. Alterations in the magnitude of the gradient in elastic modulus enhanced the directionality of invading vascular sprouts while restricting in vivo tissue invasion.
PH.D in Biomedical Engineering, December 2012
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