The Ca(2+)-induced cold gelation technique was found suitable to prepare highly porous biodegradable scaffolds based on bovine serum albumin (BSA) and alpha-casein from bovine milk for tissue engineering. A 2(3) full factorial design was used to study the influence and impact of each factor on the several responses of the scaffolds. In vitro degradation (ID), swelling ratio (SR), porosity (PO), and pore size (PS) as well cytotoxicity (CT) were evaluated and shown to be dependent on the pH of sample preparation and on the amount of BSA and casein present, making these scaffolds tunable structures. Under optimized working conditions (4.19% of BSA, 0.69% of Casein, pH 7.07), the ID attained was 37.97%, the SR observed was 11.87, the PO was 82.11%, the PS measured was 180.63 μm at surface, and 175.91 μm at fracture, whereas maximum cell viability was 84% in comparison to controls. Moreover, the scaffold supported cell adhesion and proliferation. These results, consistent with the prediction by the experimental design approach, support the use of this methodology to develop tunable scaffolds for tissue engineering using the Ca(2+)-induced cold gelation.
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