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  • Methods for photocrosslinking alginate hydrogel scaffolds with high cell viability.

Methods for photocrosslinking alginate hydrogel scaffolds with high cell viability.

Tissue engineering. Part C, Methods (2010-08-14)
Andrew D Rouillard, Caroline M Berglund, Jae Youn Lee, William J Polacheck, Yvonne Tsui, Lawrence J Bonassar, Brian J Kirby
ABSTRACT

Methods for seeding high-viability (>85%) three-dimensional (3D) alginate-chondrocyte hydrogel scaffolds are presented that employ photocrosslinking of methacrylate-modified alginate with the photoinitiator VA-086. Comparison with results from several other photoinitiators, including Irgacure 2959, highlights the role of solvent, ultraviolet exposure, and photoinitiator cytotoxicity on process viability of bovine chondrocytes in two-dimensional culture. The radicals generated from VA-086 photodissociation are shown to be noncytotoxic at w/v concentrations up to 1.5%, enabling photocrosslinking without significant cell death. The applicability of these photoinitiators for generating 3D tissue-engineered constructs is evaluated by measuring cell viability in 3D constructs with aggregate moduli in the 10-20 kPa range. Hydrogels with encapsulated bovine chondrocytes were constructed with >85% viability using VA-086. While the commonly used Irgacure 2959 is noncytotoxic in its native state and crosslinks the alginate at weight fractions much lower than VA-086, the cytotoxicity of IRG2959's photogenerated radical leads to viabilities below 70% in the conditions tested.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Alginate methacrylate, medium viscosity, degree of methacrylation: 10-30%
Sigma-Aldrich
Alginate methacrylate, high viscosity, degree of methacrylation: 20-40%
Sigma-Aldrich
Methacrylated Alginate, Medium Viscosity, Low endotoxin