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The development of large-scale human liver scaffolds equipped with interconnected flow channels in three-dimensional space offers a promising strategy for the advancement of liver tissue engineering. Tissue-engineered scaffold must be blood-compatible to address the demand for clinical transplantable liver tissue. Here, we demonstrate the construction of 3-D macro scaffold with interconnected flow channels using the selective laser sintering (SLS) fabrication method. The accuracy of the printed flow channels was ensured by the incorporation of polyglycolic acid (PGA) microparticles as porogens over the conventional method of NaCl salt leaching. The fabricated scaffold was populated with Hep G2, followed by endothelization with endothelial cells (ECs) grown under perfusion of culture medium for up to 10 days. The EC covered scaffold was perfused with platelet-rich plasma for the assessment of hemocompatibility to examine its antiplatelet adhesion properties. Both Hep G2-covered scaffolds exhibited a markedly different albumin production, glucose metabolism and lactate production when compared to EC-Hep G2-covered scaffold. Most importantly, EC-Hep G2-covered scaffold retained the antiplatelet adhesion property associated with the perfusion of platelet-rich plasma through the construct. These results show the potential of fabricating a 3-D scaffold with interconnected flow channels, enabling the perfusion of whole blood and circumventing the limitation of blood compatibility for engineering transplantable liver tissue.

Tissue engineering approaches have been clinically tried to repair damaged articular cartilages. It is an essential step to uniformly seed chondrocytes into 3D scaffolds in order to reconstruct tissue-engineered cartilages in vitro, but the tissue engineering could not have been provided with efficient cell seeding methods. Type I collagen is clinically used and known as a cytocompatible material, having recognition sites for integrins. Collagen gel encapsulating chondrocytes has been tried for making regenerated cartilages, but it is found difficult to have the gel keep its original shape after long-term culture, because of shrinking. On the other hand, 3D scaffolds, either of a nonwoven structure or a sponge-like structure, involve difficulty in that chondrocytes could not be uniformly seeded, although they have adequate initial mechanical properties. In this study, by combining collagen gelation with a nonwoven PLLA scaffold, we achieved uniform cell seeding into the 3D scaffold. Bovine articular chondrocytes were mixed with type I collagen solution, and the solution was poured into the nonwoven PLLA scaffold (1.5 mm thick, f 15 mm). The collagen–chondrocyte mixture was made into gel at 37°C for 1 h. The 0.39% collagen mixture was viscous enough to prevent cells from precipitating during gelation. Almost all chondrocytes were able to be incorporated into the PLLA scaffolds by mixing with collagen solution and subsequently making into gel, while 30–40% of the chondrocytes seeded as a cell suspension were not trapped into the PLLA scaffolds. The method presented, where chondrocytes were mixed with collagen solution, and the mixture was incorporated into a 3D scaffold, then made into gel in the scaffold, could serve as an alternative for in vitro cartilage regeneration, also simultaneously having the advantages of both materials.

To enhance the liver-specific functions of rat hepatocyte aggregates without the addition of exogenous growth factors, polylactic acid-polyglycolic acid (PLGA)/gelatin microcapsules that release insulin, dexamethasone, epidermal growth factors, and glucagon were prepared and incorporated into the hepatocyte aggregates in suspension culture. Precoating the capsules with fibronectin enhanced the incorporation of the microcapsules into the hepatocyte aggregates. In a growth factor- and hormone-free culture medium, these microcapsule-containing aggregates showed a sustained cell number and an ammonium detoxification capacity compared with two types of control culture. One was the culture of microcapsule-free aggregates with albumin-containing control capsules and the other was the culture of capsule-free aggregates that were supplied with the same factors and hormones from the culture medium at concentration levels expected from the release kinetics of the microcapsules. Our new methodology demonstrates that the performance and duration of bioartificial liver systems can be enhanced due to a more efficient maintenance of cell number by using such growth factor- and hormone-releasing microcapsules.