Barrier membranes are used in periodontal tissue engineering for successful neo‐bone tissue formation and prevention of bacterial colonization. We aimed to prepare and characterize novel 7% boron‐modified bioactive glass (7B‐BG) containing bilayered membrane for this end. We hypothesized that presence of 7B‐BG could promote structural and biological properties of guided bone regeneration (GBR) membrane. Cellulose acetate (CA) layer was prepared by solvent casting, and functionally graded layer of CA/gelatin/BG nanoparticles was prepared by electrospinning. 0B‐BG, and 7B‐BG were synthesized by quick alkali‐mediated sol–gel method and were characterized by scanning electron microscopy (SEM) and Fourier‐transform Raman spectroscopy. Membranes were cross‐linked with glutaraldehyde to preserve their stability. SEM analysis showed the asymmetric nature of membranes consisting of a smooth membrane layer and a rough surface composed of 0B‐BG and 7B‐BG containing nanofibres. 7B‐BG addition increased surface wettability (from 110.5° ± 0.8 to 73.46° ± 7.6) and biodegradability of the membranes. Additionally, a significant increase in Ca–P layer formation was observed in 7B‐BG containing group after 1‐week incubation in stimulated body fluid. 7B‐BG incorporation resulted in a decrease in tensile strength and Young's modulus values. Human dental pulp stem cells showed better attachment, spreading, and proliferation on 7B‐BG containing bilayered membranes. Osteogenic differentiation analysis revealed higher alkaline phosphatase (ALP) enzyme activity of cells (~1.5‐fold), higher intracellular calcium deposition (approximately twofold), and higher calcium deposition revealed by Alizarin red staining on 7B‐BG containing bilayered membranes. Overall, results suggested that functionally graded bilayered membranes hold potential for GBR applications in regenerative dentistry.
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