Polyhydroxybutyrate (PHB) is a biopolymer produced by bacteria. This study aimed to implement the production process of experimental medical-grade PHB and to evaluate its morphology and biocompatibility compared to conventional resin-based composites (RBCs). PHB raw material was produced via biological process and then the membrane was generated via electrospinning specifically for this study and imaged with Micro-Computed Tomography (Micro-CT) and scanning electron microscopy (SEM). MTS assay was used to assess the cytotoxicity of PHB compared to other materials. Test groups included two packable resin composites (Point 4-Kerr, G-aenial anterior-GC), two flowable resin composites (Filtek Ultimate Flowable-3M ESPE, Nova Compo HF-Imicryl), a compomer (Nova Compomer-Imicryl), a fissure-sealant (Fissured Nova Plus-Imicryl), and the PHB membrane (Innovaplast Biotechnology Inc., Eskisehir, Turkey). A control group consisting of cells without any test material was also produced. To perform the MTS assay, disc-shaped specimens of the aforementioned materials were prepared and then incubated with mouse fibroblast cells (L929) for 24 and 48 h. Micro-CT and SEM images revealed a homogeneous and fibrillary structure of the PHB. MTS assay revealed the highest cell viability in the PHB, Nova Compomer, and Fissured Nova Plus groups after 24 h. PHB and Nova Compomer showed the highest viability rates at 48 h while other RBCs had rates below 25% (