Abstract : strongBACKGROUND: /strongCurrent resorbable and non-resorbable membranes act as a physical barrier to avoid connective and epithelial tissue downgrowth into the defect, favoring the regeneration of periodontal tissues. These conventional membranes possess many structural and bio-functional limitations. We hypothesized that the next-generation of guided tissue regeneration (GTR) membranes for periodontal tissue engineering will be a biologically active, spatially designed nanofibrous biomaterial that closely mimics the native extra-cellular matrix (ECM).strongMETHODS: /strongGTR membranes made of poly(ε-Caprolactone) with a molecular weight of 80,000 reinforced with different weight concentrations of nano-Hydroxyapatite/Bioactive glass (2%, 5%, 10%, 15%) is fabricated by the method of electrospinning. After fabrication, in vitro properties are evaluated.strongRESULTS: /strongThe electrospun nanofibrous membranes possessed excellent mechanical properties initially and after one month of degradation in phosphate buffer solution (PBS). Moreover, none of the fabricated membranes were found to be cytotoxic at lower concentrations and higher concentrations. Comparing the overall properties, PCL (poly(e-caprolactone)) + BG (Bioactive glass) 2% exhibited superior cell attachment and percentage of viable cells, increased fiber and pore diameter which satisfies the ideal properties needed for GTR membranes.strongCONCLUSION: /strongComposite nanofibrous membranes prepared by electrospinning are suitable for use as a GTR membrane and are a useful prototype for further development of a final membrane for clinical use.