Anisotropic temperature sensitive chitosan-based injectable hydrogels mimicking cartilage matrix

In this study, chitosan-based hydrogels were formulated with material similarities to three of the four zones of articular cartilage. Gelatin, hyaluronic acid (HA), and β-tricalcium phosphate for the superficial, radial, and calcified zones, were blended in different amounts and tested for formation of uniform solution, gelability, and rheological characteristics. Confined compression in two configurations (series and parallel to anisotropy), and cyclical tests were performed at the physiological conditions. In vivo gelation and systemic effects were evaluated in male BALB/c mice subcutaneous model. At day 5, hydrogels were harvested along with the adjoining skin and analyzed by histology. Formulations that produced solutions after pH adjustments were selected for each zone. Anisotropic hydrogels were formed by mixing solutions from each zone, which showed uniform gradation. Addition of HA improved structural integrity relative to other formulations. When hydrogels were in series, combined hydrogel modulus was the average of all zones while that in parallel orientation was half of that series orientation. Cyclical tests demonstrated repeatable strength and durability. All formulations were injectable into the subcutaneous region. H/E stained tissues showed minimal invasion of inflammatory cells in radial and calcified zones. Structural integrity of the hydrogel is suggested to be the resultant of the presence of HA.