Three-Dimensional Composite of Fish Collagen Peptides and Bacterial Cellulose Promotes HT-22 in vitro neurite outgrowth

Background: Bacterial cellulose (BC) is a highly biocompatible biopolymer valued for its unique nanofibrillar structure, excellent mechanical strength, high water retention, and intrinsic non-toxicity, making it particularly suitable for biomedical applications.Methods: A three-dimensional (3D) composite scaffold composed of BC and fish collagen peptides (FCP) was fabricated via a one-step in situ biosynthesis method by incorporating optimized concentrations of FCP (0.1% and 0.5%) into the BC culture medium during bacterial fermentation. The structural integrity and surface morphology of the scaffolds were examined using field emission scanning electron microscopy (FE-SEM). The biological functionality of the scaffolds was further evaluated by culturing HT-22 neuronal cells (mouse hippocampal origin) on both pristine BC and BC-FCP scaffolds. Cell adhesion, morphology, and neurite outgrowth were evaluated using fluorescence microscopy after 2 days of incubation.Results: The BC-FCP composite scaffolds demonstrated superior microstructural integrity and biological performance compared to pristine BC. Field-emission scanning electron microscopy revealed a denser and more uniform nanofibrous architecture in BC-FCP scaffolds, confirming the successful incorporation and uniform distribution of collagen peptides within the cellulose matrix. HT-22 neuronal cells cultured on these scaffolds showed markedly enhanced adhesion, spreading, and neurite outgrowth, particularly at the higher FCP concentrations (0.5%) demonstrating the neuro-supportive capability of the composite system.Conclusion: The BC-FCP composite scaffolds significantly enhance neuronal adhesion and growth, making them promising candidates for neural tissue engineering and regenerative applications.Keywords:Bacterial cellulose, Fish collagen peptides, 3D scaffolds, Biocompatibility, Neuronal outgrowth