Preparation of chitosan-gelatin hybrid scaffolds with well-organized microstructures for hepatic tissue engineering.

The structural organization of natural liver is instrumental in the multifunctionality of hepatocytes, and mimicking these specific architectures in tissue-engineered scaffold plays an important role in the engineering of an implantable liver equivalent in vitro. To achieve this goal, we have developed a novel fabrication process to create chitosan-gelatin hybrid scaffolds with well-organized architectures and highly porous structures by combining rapid prototyping, microreplication and freeze-drying techniques. The scaffolds obtained not only have analogous configurations of portal vein, central vein, flow-channel network and hepatic chambers, but also have high (>90%) porosity, with the mean pore size of 100microm. Swelling and degradation studies showed that the scaffold has excellent properties of hydrophilicity and biodegradability. A hepatocyte culture experiment was conducted to evaluate the efficiency of the well-defined chitosan-gelatin scaffold in facilitating hepatocyte growth in the inner layer of the scaffold in vitro. Scanning electron microscopy and histological analysis showed that hepatocytes could form large colonies in the predefined hepatic chambers, and these cavities could the completely filled with hepatocytes during 7 day culture. Albumin secretion and urea synthesis further indicated that the well-organized scaffolds were more suitable for hepatocyte culture.