Characterization of bacterial nanocellulose produced by isolates from Philippine nata starter and its biocompatibility

Abstract

Two nanocellulose-producing bacteria were isolated from separate local nata starter cultures and were characterized to determine their putative identity. The nanocellulose produced by the two bacterial isolates were compared to determine if differences between material properties and in vitro biocompatibility were species-dependent. Nanocellulose membranes were produced by inoculating the bacterial isolates to Hestrin-Schramm media using different carbon sources (glucose, mannitol and sucrose). The material properties of nanocellulose membranes were characterized using scanning electron microscopy, Fourier-transform infrared spectroscopy, X-ray dispersive spectroscopy, Calcofluor staining and water retention. The biocompatibility of the nanocellulose membranes was evaluated using MC3T3 pre-osteoblast cells and was analyzed using CCK-8 cell viability assay. Adhering cells were stained and viewed under confocal laser scanning microscope. A selected sample was implanted subcutaneously in Sprague Dawley rats for one week and four weeks to determine its in vivo biocompatibility. Hematoxylin and eosin staining of the subcutaneous tissue sections with implant was examined to visualize host cell behavior to the implant.