〔Abstract〕 To prepare silk protein (SF) membranes reinforced by aragonite sheets (AP) and to inves- tigate their potential as novel barrier membranes for bone tissue regeneration by testing their tensile properties , bio- compatibility , and their effect on osteogenic differentiation . Methods AP was extracted from natural abalone shell (AS) by oxidation method , and AP-SF composite and pure SF membranes with different component ratios were pre- pared by solution casting and evaporative self-assembly techniques , and their tensile properties were tested using a universal mechanical machine under wet state conditions . The tensile properties were tested using a universal me- chanical machine under wet conditions . The microstructure of the AP-SF membrane with the highest tensile strength was observed using scanning electron microscopy (SEM) and characterized using Fourier transform infrared spec- troscopy (FTIR) and X-ray diffraction (XRD) . Observation of the adhesion of rat bone marrow mesenchymal stem cells (rBMSCs) on the surface by scanning electron microscopy , and exploration of the biocompatibility of AP-SF membrane by CCK-8 and live cell staining. Alkaline phosphatase staining (ALP) and alizarin red staining (ARS) were used to detect the osteogenic differentiation of rBMSCs in each group . Results AP was successfully extracted from AS , and the membrane prepared from a mixed solution of SF and AP with a solid content of 10 ∶9 had the strongest wet tensile performance , reaching 8. 46 MPa. The CCK-8 and live dead cell staining test results of the AP-SF membrane showed no significant difference compared to the blank group and SF group , indicating that both membranes had good biocompatibility. The ALP test and ARS results indicated that AP-SF membrane had a more significant ability to promote osteogenic differentiation of rBMSCs compared to SFmembrane . Conclusion The prepared AP-SF membrane has both good mechanical and biological properties , and has certain clinical application potential .