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Novel bilayer bacterial nanocellulose scaffold supports neocartilage formation in vitro and in vivo
- Martínez Ávila, Héctor, Feldmann, Eva-Maria, Pleumeekers, Mieke M., Nimeskern, Luc, Kuo, Willy, de Jong, Willem C., Schwarz, Silke, Müller, Ralph, Hendriks, Jeanine, Rotter, Nicole, van Osch, Gerjo J.V.M., Stok, Kathryn S., Gatenholm, Paul
- Biomaterials 2015 v.44 pp. 122-133
- alginates, biocompatibility, biomechanics, bone marrow, cartilage, cell viability, cellulose, chondrocytes, endotoxins, humans, immunohistochemistry, mice, morphometry, porosity, porous media, tissue engineering
- Tissue engineering provides a promising alternative therapy to the complex surgical reconstruction of auricular cartilage by using ear-shaped autologous costal cartilage. Bacterial nanocellulose (BNC) 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 BNC scaffold for auricular cartilage tissue engineering. Bilayer BNC scaffolds, composed of a dense nanocellulose layer joined with a macroporous composite layer of nanocellulose and alginate, were seeded with human nasoseptal chondrocytes (NC) and cultured in vitro for up to 6 weeks. To scale up for clinical translation, bilayer BNC scaffolds were seeded with a low number of freshly isolated (uncultured) human NCs combined with freshly isolated human mononuclear cells (MNC) from bone marrow in alginate and subcutaneously implanted in nude mice for 8 weeks. 3D morphometric analysis showed that bilayer BNC scaffolds have a porosity of 75% and mean pore size of 50 ± 25 μm. Furthermore, endotoxin analysis and in vitro cytotoxicity testing revealed that the produced bilayer BNC scaffolds were non-pyrogenic (0.15 ± 0.09 EU/ml) and non-cytotoxic (cell viability: 97.8 ± 4.7%). This study demonstrates that bilayer BNC scaffolds offer a good mechanical stability and maintain a structural integrity while providing a porous architecture that supports cell ingrowth. Moreover, bilayer BNC scaffolds provide a suitable environment for culture-expanded NCs as well as a combination of freshly isolated NCs and MNCs to form cartilage in vitro and in vivo as demonstrated by immunohistochemistry, biochemical and biomechanical analyses.