Biofabrication of bacterial cellulose and hydroxylated hexagonal boron nitride nanocomposites based on in situ production strategies: Characterization and cytotoxicity evaluation

Bacterial cellulose is considered a highly promising biomaterial due to its three-dimensional network structure, especially its nanometer-scale fibers, which closely resemble the natural extracellular matrix. Hexagonal boron nitride is a novel material that has attracted increasing interest in the field of biomedicine. Hydroxylated hexagonal boron nitride modified bacterial cellulose (BC/BN) nanocomposite was produced by an in situ fermentation method. The physicochemical properties of the composite hydrogels were investigated by Fourier transform infrared spectroscopy, x-ray diffraction, thermogravimetric analysis, and scanning electron microscopy (SEM). For tissue application, the response of human osteoblast cells on the BC/BN nanocomposites has been evaluated in this study. The chemical structure, morphology, and hydrophilicity of the nanocomposites were characterized as well as, and the viability, adhesion and morphology of cells were also investigated. Cell proliferation, morphology, and adhesion were evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) proliferation assays, scanning electron microscopy, and confocal microscopy, respectively. The results show that BC/BN nanocomposites promote higher levels of cell viability, adhesion, and proliferation compared to BC. The incorporation of BN was increased cell viability and enhanced osteoblast cells adhesion. These results demonstrate the potential application of the BC/BN nanocomposite for cellular response studies. This study aims to create a new generation of bio-nanocomposite scaffolds. To create mechanically reinforced, biocompatible, and renewable matrices that solve important problems in tissue engineering and regenerative medicine, this study aims to combine sustainable biological production with functional nanomaterials. In this regard, hydroxylated hexagonal boron nitride (BN) has been incorporated into bacterial cellulose (BC) through an in situ biofabrication process. The in situ approach developed in this study offers an environmentally friendly and scalable method for directly embedding BN nanolayers into the cellulose network during fermentation. This method eliminates the need for harsh post-processing chemicals used during the production of hydroxylated hexagonal boron nitride, thereby reducing production costs and increasing structural homogeneity, offering a suitable pathway for the industrial-scale biofabrication of multifunctional biomaterials. Furthermore, this method not only produces hydroxylated hexagonal boron nitride but also enables its integration with membranes in a single step. Compared to pure bacterial cellulose material, the resulting BC/BN nanocomposites exhibit enhanced hydrophilicity, thermal stability, and cell viability. This enables the production of mechanically reinforced, biocompatible, and renewable functional nanomaterials that solve significant problems in tissue engineering and regenerative medicine through sustainable biological production. The in situ approach developed in this study offers an environmentally friendly and scalable method for directly embedding BN nanolayers formed during fermentation into the cellulose network. This method eliminates the need for harsh post-treatment chemicals, reduces production costs, and increases structural homogeneity, offering a viable pathway for the industrial-scale biological production of multifunctional biomaterials.