Orthotopic liver transplantation is presently the most effectual method for the treatment of end-stage liver diseases. Though, one major issue is the restricted number of donor organs that are accessible. Hence, liver tissue engineering is under investigation with the goal of restoring liver functions. In this study, we investigated 3D porous scaffolds made of PLLA coated with a nano thick collagen layer (matrices). Primary rat dermal fibroblasts were used in a first study phase to check matrices' cytocompatibility. More than 70% of seeded cells could adhere and remain viable 24 and 48 hours after the seeding. To test the suitability of the matrices for human primary hepatocytes, HepaRG cells were seeded and analyzed for viability, adhesion rate, and functionality such as albumin secretion. About 80% of seeded HepaRG adhered to the scaffolds remaining viable up to 72 hours. Cells were homogeneously distributed in the entire scaffold with albumin secretion increasing with time. Our results indicate that PLLA collagen-coated matrices allow hepatocytes attachment and distribution throughout the 3D structure, as well as support cell functionality. Such matrices have been applied in our clinical phase II trial. Functional hepatocytes were successfully implanted in patients suffering from liver-cirrhosis with higher cell numbers and adhesions rate compared to our previous trial with the first matrix type and a general improvement in clinical condition.
Keywords: 3D PLLA scaffolds; hepatocyte; nano-thick collagen coating; tissue engineering.
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