Multilayered polyvinyl alcohol-based nanofibrous membranes functionalized for osteogenic induction of mesenchymal stem cells

Bone scaffolds require osteogenic differentiation and mechanical integrity. This study developed a multilayered nanofiber membrane (MNFM) composed of PVA–ZnO, PVA–COL I–BMP2, and PVA–HA via electrospinning to mimic bone matrix. Fourier Transform Infrared Spectroscopy (FTIR) and Energy-Dispersive X-ray Spectroscopy (EDS) analyses confirmed the successful incorporation of ZnO, COL I, and HA. The nanofibers showed random orientation and excellent hydrophilicity. Mechanical tests revealed a UTS of 19 MPa and elastic modulus of 24.7 MPa. MNFM exhibited rapid swelling (~ 250%) and controlled degradation (~ 60% mass loss at 48 h). In vitro studies with hBMMSCs showed enhanced adhesion, proliferation, and osteogenic differentiation. RT-qPCR analysis revealed increased expression of BSP, OCN, OPN, and ONN over time. Western blotting confirmed upregulated COL1A1, RUNX2, and OSX, indicating support for extracellular matrix formation and early osteogenesis. Elevated ALP and calcium levels suggested late-stage differentiation.