Construction of nacre-like structure-enhanced silk protein GBR membrane and its performance in vitro experiments

Acta Universitatis Medicinalis Anhui 2025, 05, v.60 906-912     font:big middle small

Found programs: Health Research Project of Anhui Province(No.AHWJ2022b001);Research Project of Anhui Provincial Institute of Translational Medicine(No.2022zhyx-B03)

Authors:Yang Feiyu; Xu Wenhua

Keywords:aragonite platelets;silk fibroin;nacre;guided bone regeneration;bone defect repair;osteogenic differentiation

DOI:10.19405/j.cnki.issn1000-1492.2025.05.018

〔Abstract〕 Objective To prepare silk protein(SF) guided bone regeneration(GBR) membranes reinforced by aragonite sheets(AP) and to investigate their potential as novel barrier membranes for bone tissue regeneration by testing their tensile properties,biocompatibility,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 prepared 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 mechanical 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 spectroscopy(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 SF membrane.Conclusion The prepared AP-SF membrane has both good mechanical and biological properties,and has certain clinical application potential.