Mineralized collagen-glycosaminoglycan scaffolds were fabricated via lyophilization from a mineralized collagen precursor suspension as described before.47 (link),65 ,66 The precursor suspension was created by homogenizing type I collagen (1.9 weight per volume, Sigma Aldrich, St. Louis, Missouri USA), chondroitin-6-sulfate (0.84 weight per volume, Sigma Aldrich), and calcium salts (calcium hydroxide and calcium nitrate, Sigma Aldrich) in a mineral buffer solution (0.1456 M phosphoric acid/0.037 M calcium hydroxide). The precursor suspension was stored at 4 °C and degassed prior to lyophilization.
Mineralized collagen scaffolds were fabricated via lyophilization using a Genesis freeze-dryer (VirTis, Gardener, New York USA) as described before.66 Briefly, 100 μL of precursor suspension was pipetted into a custom 144-well polysulfone mold (6 mm diameter, 7 mm tall wells). The precursor solution was frozen by cooling from 20 °C to −10 °C at a constant rate of 1 °C per minute followed by a temperature hold at −10 °C for 2 hours. The frozen suspension was then sublimated at 0 °C and 0.2 Torr, resulting in a porous scaffold network.
All scaffolds were hydrated for 2 hours in ethanol, cross-linked for 2 hours in EDC-NHS, and washed in phosphate buffered saline (PBS) for 48 hours prior to use in experiments.
Mineralized collagen scaffolds were fabricated via lyophilization using a Genesis freeze-dryer (VirTis, Gardener, New York USA) as described before.66 Briefly, 100 μL of precursor suspension was pipetted into a custom 144-well polysulfone mold (6 mm diameter, 7 mm tall wells). The precursor solution was frozen by cooling from 20 °C to −10 °C at a constant rate of 1 °C per minute followed by a temperature hold at −10 °C for 2 hours. The frozen suspension was then sublimated at 0 °C and 0.2 Torr, resulting in a porous scaffold network.
All scaffolds were hydrated for 2 hours in ethanol, cross-linked for 2 hours in EDC-NHS, and washed in phosphate buffered saline (PBS) for 48 hours prior to use in experiments.
