〔Abstract〕 Objective To explore the feasibility of silk fibron(SF)-phlorotannins(PT) double layer membranes as barrier membrane for guided bone regeneration(GBR) by exploring the mechanical property, biocompatibility, and the effect on osteogenic differentiation of SF-PT double layer membranes. Methods The SF solution treated with different degumming time was prepared into SF bilayer by solution cast technique, ice template method and lyophilization. The tensile strength of the membranes under hydration were measured via mechanical universal machine(n=5). The microstructures of the composite membranes with the highest tensile strength under hydration condition were observed by scanning electron microscopy. Then the rough surfaces of SF double layer membranes was loaded with PT to prepare SF-PT double layer membranes, Fourier transformed infrared spectroscopy was carried out to characterize the interaction between PT and SF double layer membranes,and the bone marrow mesenchymal stem cells of human alveolar origins( h ABMSCs) adhesion was studied by SEM. CCK-8 assay and live dead staining were used to evaluated the biocompatibility of SF-PT double layer membranes. qRT-PCR,ALP staining and alizarin red stain were carried out to measure the osteogenic differentiation of h ABMSCs in different groups. Results The SF membranes prepared from SF solution with a degumming time of 30 min has the highest tensile strength,reaching( 3. 293 ± 0. 122 8) MPa. The smooth surface of SF double layer membrane was dense and smooth,while the rough surface was longitudinally ordered loose porous structure. PT was loaded onto the rough surface of the SF double layer membranes by hydrogen bond,hydrophobic interaction and other forces. There was no significant difference in the results of CCK-8 and live dead staining between blank group,SF group and PT-SF group. Compared with blank group and SF group,PT-SF double layer membranes promoted osteogenic differentiation of h ABMSCs. Conclusion Thus,the PT-SF double layer membranes expressed desired mechanical,biological properties and can promote the osteogenic differentiation of h ABMSCs to a certain extent. These results indicate that PT-SF double layer membranes is successfully fabricated,providing a solid foundation for future in vivo experiments.