15502-m-pleumeekers

ABSTRACT Tissue engineering provides a promising alternative therapy to the complex surgical reconstruction of auricular cartilage by using ear-shaped autologous costal cartilage. Bacterial nanocellulose is proposed as a promising scaffold material for auricular cartilage reconstruction, as it exhibits excellent biocompatibility and secures tissue integration. Thus, this study evaluates a novel bilayer bacterial nanocellulose scaffold for auricular cartilage tissue engineering. Bilayer bacterial nanocellulose scaffolds, composed of a dense nanocellulose layer joined with a macroporous composite layer of nanocellulose and alginate, were seeded with human nasoseptal chondrocytes and cultured in vitro for up to 6 weeks. To scale up for clinical translation, bilayer bacterial nanocellulose scaffolds were seeded with a low number of freshly isolated (uncultured) human nasoseptal chondrocytes combined with freshly isolated human mononuclear cells from bone marrow in alginate and subcutaneously implanted in nude mice for 8 weeks. 3D morphometric analysis showed that bilayer bacterial nanocellulose scaffolds have a porosity of 75% and mean pore size of 50 ± 25 mm. Furthermore, endotoxin analysis and in vitro cytotoxicity testing revealed that the produced bilayer bacterial nanocellulose scaffolds were non-pyrogenic (0.15 ± 0.09 EU/ml) and non- cytotoxic (cell viability: 97.8 ± 4.7%). This study demonstrates that bilayer bacterial nanocellulose scaffolds offer a good mechanical stability and maintain a structural integrity while providing a porous architecture that supports cell ingrowth. Moreover, bilayer bacterial nanocellulose scaffolds provide a suitable environment for culture-expanded NCs as well as a combination of freshly isolated nasoseptal chondrocytes and mononuclear cells to form cartilage in vitro and in vivo as demonstrated by immunohistochemistry, biochemical and biomechanical analyses. 148 CHAPTER 8