Negative control (no hyase) Effect of on Degradation 5 u/ml hyase n>3 for each point Incubate in hyase Measure weight loss ![]() Hubbell’s laboratoriesĬonjugation Efficiency and Yield Hexaglycine input Conjugation efficiency Hexaglycine input Conjugation yield Targeted yield * * Based on work with PEG hydrogels in J. (submitted) J Biomed Mater Res * Peptides of interest: Cell adhesive fibronectin peptide (GRGDSG) Model peptide, hexaglycine (GGGGGG) * Based on work with PEG hydrogels in J. Peptide-Conjugated GMHA Hydrogels Baier Leach J, Bivens KA, Collins CN, & Schmidt CE. In Vivo Analysis of EC Infiltration Fibrin (+) GMHA hydrogel 6.63 ± 1.10, n = 9 (% area CD31-positive cells)7.06 ± 0.14, n = 4 2 Week Implant Scale, 200 mm Agarose (-) EC immunostain (CD31) In Vivo Analysis of Endothelial Cell (EC) Infiltration Fill TEC with hydrogel 4 hydrogels per rat + Control: Fibrin -Control: Agarose 2 HA gels 2. Harvest tissue at 2 weeks (side view without TEC) 1. Media Hydrogel Crosslinked GMHA 1% GMHA 0.1% Irgacure 2959 0.03% N-vinyl pyrrolidinone 1 minute UV Crosslinked GMHA GMHA in solution Media HA n>6 for each bar Effect of Methacrylation on Cytocompatibility HAEC monolayer Pore size Effect of Methacrylation on Crosslinking n=3 for each barĥ% 7% 11% Increasing % methacrylation n>4 for each point Effect of Methacrylation on Degradation Rate GMHA Solution +UV Incubate in hyase Measure weight loss of gel over time Remove mold.GMHA Solution +UV Remove mold Equilibrate in buffer overnight, weigh Dry completely, weigh Determining the Degree of Crosslinking (Swelling Ratio) Goal: To obtain a relative measure of crosslinking for the GMHA gelsįlory polymer-solvent interaction theory: Estimated pore size: 644 nm 619 nm 539 nm (2003) Biotech Bioeng 82:578-89 Crosslinking Variables: GMHA concentration (0.5-2.0%) UV exposure (1-4 min, ~22 mW/cm2) Photoinitiator conc. Glycidyl methacrylate (GM) + Photoinitiator + UV light Crosslinked GMHA GMHA HA GM modification HA Methacrylation and Crosslinking Baier Leach J, Bivens KA, Patrick CW, Jr. ![]() ![]() Overall Goal: To develop and characterize hyaluronic acid hydrogel scaffolds for soft tissue engineering applicationsĪim 1: To create and characterize HA hydrogels Aim 2: To develop methods for controlling cell-HA hydrogel interactions Peptide-conjugated HA hydrogels Protein-releasing HA hydrogelsĪim 1: To create and characterize HA hydrogels Aim 2: To develop methods for controlling cell-GMHA hydrogel interactions Peptide-conjugated GMHA hydrogels Protein-releasing GMHA hydrogels HA’s role in wound healing Chen WY & Abatangelo G. Multiple sites available for modification.Hyaluronic Acid (HA) glucuronic acid acetylglucosamine Tissue Engineering Scaffolds: State of the Art Proteins Fibrin, Collagen “Sugars” Agarose, Alginate Chitosan, Dextran, Hyaluronic acid Synthetic polymers Polyethylene glycol (PEG) Polylactic-co-glycolic acid (PLGA) Polyhydroxyethyl methacrylate (pHEMA) Inherent biological activity Nonimmunogenic Multiple modification sites Tunable material properties 78:211-8 Aim: To facilitate natural wound healing biology Biological design: Biomimetic molecules like those present in a wound Limited nonspecific protein adsorption Nonimmunogenic Materials design: Enzymatic degradation Versatile modification strategies Mechanical properties match tissue Hubbell JA. Hyaluronic Acid Hydrogel Biomaterials for Soft Tissue Engineering Applications Jennie Baier Leach Supervisor: Christine E.
0 Comments
Leave a Reply. |