Preparation, characterization and cell response studies on bioconjugated 3D protein hydrogels with wide-range stiffness: An approach on cell therapy and cell storage.

The present study emphasizes the preparation and characterization of bioconjugated keratin-gelatin (KG) 3D hydrogels with wide-range stiffness to study cell response for cell therapy and cell storage applications. In brief, human hair keratin and bovine gelatin at different ratios bioconjugated using EDC/NHS provide five hydrogels (KG-1, KG-2.5, KG -5, KG-7.5 and KG-9) with modulus ranging from 0.9 ± 0.1 to 10.9 ± 0.4 kPa. Based on swelling, stability, porosity, and degradation parameters KG-5 and KG-9 are employed to assess the human dermal fibroblast (HDF) cell response, cell delivery and cell storage respectively. Characterization studies revealed the concentration of keratin determines the modulus/stiffness of the hydrogels, whereas gelatin concentration plays a vital role in porosity, swelling percentage, and degradation properties. HDF cell behaviour in the chosen hydrogels assessed based on cell adhesion, cell proliferation, PCNA expression, MTT assay, and DNA quantification. We observed the best cell behaviour in KG-5 hydrogels than in the KG-9 matrix. In cell storage and cell delivery studies, the KG-9 matrix displayed promising results. Thus, the present study concludes bioconjugated keratin-gelatin 3D hydrogel with modulus below 3.0 kPa facilitates the proliferation of HDFs, whereas matrix above 10 kPa modulus supports cell storage and cell recovery. The observations of the present study suggest the suitability of bioconjugated fibrous protein 3D hydrogel for cell therapy and cell storage.