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BNC well integrated with the host and neo-tissue. The non-degradable BNC will provide long- term structural integrity after implantation, and has previously shown remarkable integration with the host tissue in different animal models [313, 314, 316, 361]. A novel BNC scaffold designed with a bilayer architecture that integrates mechanical stability and high porosity was successfully fabricated and evaluated for auricular cartilage TE, in vitro and in vivo . In conclusion, this study demonstrates that non-pyrogenic and non-cytotoxic bilayer BNC scaffolds can be successfully produced. Furthermore, such scaffolds, together with alginate, provide a suitable environment for culture-expanded human nasoseptal chondrocytes and freshly isolated human nasoseptal chondrocytes combined with freshly isolated human mononuclear cells to form cartilage in vitro and in vivo . Most studies that have used biodegradable materials to engineer auricular cartilage have resulted in poor structural integrity of the scaffold after implantation due to the short-lived chemical stability of the scaffold material. This study found that bilayer BNC scaffolds offer a good mechanical stability and maintain a structural integrity while providing a porous architecture that supports cell ingrowth and neocartilage formation, as demonstrated by immunohistochemistry, biochemical and biomechanical analyses. Ongoing work focuses on developing bilayer BNC scaffolds in the shape of a human auricle, aiming to provide an effective treatment to serious auricular defects. Acknowledgements This study was performed within the framework of EuroNanoMed (EAREG-406340-131009/1) and was supported by the Swedish Research Council (2009-7838), Federal Ministry of Education and Research (13N11076), SenterNovem (ENM09001) and the Swiss National Science Foundation (NRP63). The authors acknowledge dr. Annette Jork and CellMed AG (Alzenau, Germany) for providing medical grade alginate; Athanasios Mantas and Anders Mårtensson at Chalmers University of Technology (Gothenburg, Sweden) for assistance with scaffold production and FTIR analysis, respectively; and Priscila Martínez Ávila at The University of Texas at Arlington for helping with statistical analysis. The antibody used for immunohistochemical type II collagen detection (II-II6B3) was obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at the University of Iowa, Department of Biology, Iowa City, IA 52242, USA. 170 CHAPTER 8