Colorimetric assessment of biomolecule incorporation in hydrogels

Colorimetric assays were purchased from Thermo Fisher Scientific (Waltham, MA) and used according to the manufacturer’s instructions. To quantify the degree of incorporation of biomolecule incorporation in hydrogels, the hydrogels were leached in 1.5 ml of PBS at pH 7.4 for 24 hours at 37°C, and then, the surrounding solution was collected. The liquid solutions containing the hydrogel sol fractions were then quantified for CS, BMPm, or NC concentration using a DMMB kit for sulfated GAGs or a Coomassie blue kit for peptides (32, 33). Following this, the percentage of biomolecule retention in gels was computed by dividing the eluted concentration by the original biomolecule concentration and subtracting from 100%. Standard curves for both assays can be found in the Supplementary Materials.

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