Objective: To explore the feasibility of graphene oxide (GO)-chitosan (CS) guided bone regeneration composite membrane being used as a new guided bone regeneration (GBR) membrane by testing its tensile strength and its effect on the proliferation of human gingival fibroblasts (HGF). Methods: The CS solution and GO solution were mixed in following ratios, ultrasonically crushed and dispersed, and formed into membranes by self-evaporation. Composite membrane with 0.5%, 1.0%, 2.0%, 4.0%, 6.0%, 8.0% GO in CS was prepared. The tensile strength of the composite membranes was tested by mechanical universal testing machine (n=6). The microstructure of the composite membrane with the best tensile strength was observed by scanning electron microscopy and transmission electron microscopy. X-ray diffraction was used to characterize the ingredients of the material. A group of samples was prepared again with the proportion of the highest tensile strength composite membrane, and were immersed in a sodium hydroxide solution for acid removal. The tensile strength of the new group of samples was tested. The newly extracted impacted teeth were collected from the Department of Oral and Maxillofacial Surgery, Stomatologic Hospital and College, Anhui Medical University, and the gingival tissues remained on the teeth were taken for primary HGF and were cultured to P2 generation, which were identified by immunocytochemical staining methods. Cell counting kit-8 (CCK-8) was applied to detect cell counts in the blank control group, the pure CS membrane group, and the composite membrane group with the highest tensile strength (n=5) and all groups co-cultured with the HGF for 24 h and 48 h. Results: After adding GO, the cross-section of the composite membrane was in an ordered layer structure. X-ray diffraction analysis showed that GO could be found in the composite membrane. The tensile strength increased with the increase of GO ratio. When the mass fraction of GO in CS was 4.0%, the tensile strength reached (134.8±7.3) MPa and the tensile strength reached (144.6±8.1) MPa after deacidification of the composite membrane. The CCK-8 test of HGF showed that there was no significant difference in absorbance between the pure CS membrane group and the GO/CS composite membrane group when they were compared with the blank control group (P>0.05). Conclusions When the mass fraction of GO in CS is 4.0%, the composite membrane has the best tensile strength. The composite membrane showes good cytocompatibility, which lays a foundation for further in vivo experiments and the development of a new generation of GBR membrane.