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Immunostaining of cartilage matrix proteins within each zone of dual-gradient hydrogel. Increasing CS and stiffness in dual-gradient hydrogels led to larger nodules of neocartilage and more ECM matrix deposition with minimal hypertrophic phenotype and fibrocartilage observed. The encapsulated cells retained a rounded morphology in stiffer/high-CS zones (i.e., zone 4/5) while the cells present in softer/low-CS zones (i.e., zone 1/2) became elongated and spread. Scale bar = 50 μm.
