Abstract
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.
Highlights
Mature cartilage has a very limited ability for repair, which has led to intense research toward the development of cell-seeded scaffolds
Our results clearly showed that Bacterial nano-cellulose (BNC) secreted by mutant strain preserved natural biocompatibility of BNC and served as superior mammalian cells support both in terms of chondrogenic cells proliferation, morphology and extracellular matrix secretion (GAG production level estimated with Alcian blue staining) when compared to membranes produced by the wild-type strain
The cellulose membranes produced by this strain are more gel-like when compared to other effective BNC producers, e.g., K. hansenii ATCC 53582
Summary
Mature cartilage has a very limited ability for repair, which has led to intense research toward the development of cell-seeded scaffolds. Scaffolds provide a 3D surrounding for cells what prevents dedifferentiation of chondrocytes into a fibroblast-like cell type, what has been first shown for agarose gels [2]. Even slight changes in the structure of a material supporting differentiating chondrocytes can influence their fate, as recent research on electro-spun synthetic scaffolds indicated [3,4]. The material should provide mechanical support in order to accurately recreate the natural tissue endurance and to generate transportable products for clinical use [8]. A perfect scaffold material for cartilage regeneration should be moldable in order to fill the defect in natural tissue in vivo and, beforehand, to properly guide proliferating cells in vitro
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