Development of three-dimensional scaffolds mimicking in vivo cells' environment is an ongoing challenge for tissue engineering. Bacterial nano-cellulose (BNC) is a well-known biocompatible material with enormous water-holding capacity. However, a tight spatial organization of cellulose fibers limits cell ingrowth and restricts practical use of BNC-based scaffolds. The aim of this study was to address this issue avoiding any chemical treatment of natural nanomaterial. Genetic modifications of Komagataeibacter hansenii ATCC 23769 strain along with structural and mechanical properties characterization of obtained BNC membranes were conducted. Furthermore, the membranes were evaluated as scaffolds in in vitro assays to verify cells viability and glycosaminoglycan synthesis by chondrogenic ATDC5 cells line as well as RBL-2H3 mast cells degranulation. K. hansenii mutants with increased cell lengths and motility were shown to produce BNC membranes with increased pore sizes. Novel, BNC membranes with relaxed fiber structure revealed superior properties as scaffolds when compared to membranes produced by a wild-type strain. Obtained results confirm that a genetic modification of productive bacterial strain is a plausible way of adjustment of bacterial cellulose properties for tissue engineering applications without the employment of any chemical modifications.
Keywords: 3D culture; Komagataeibacter; bacterial cellulose fiber structure; bacterial nano-cellulose; chondrogenic differentiation; scaffold.