To　improve　bone　regeneration　in　oral　microenvironment,　we　generated　a　novel　biodegradable,　antibacterial,　and　osteoconductive　electrospun　PLGA/PCL　membrane　as　an　ideal　osteogenic　scaffold.　The　novel　three-layer　membranes　were　structured　with　serial　layers　of　electrospun　chlorhexidine-doped-PLGA/PCL　(PPC),　PLGA/PCL　(PP),　and　beta-tricalcium　phosphate-doped-PLGA/PCL　(PP　beta).　To　characterize　osteoconductive　properties　of　these　membranes,　MC3T3-E1　(MC)　cultures　were　seeded　onto　the　membranes　for　14　days　for　evaluation　of　cell　proliferation,　morphology　and　gene/protein　expression.　In　addition,　MC　cells　were　cultured　onto　different　surfaces　of　the　three-layer　membranes,　PPC　layer　facing　MC　cells　(PP　beta-PP-PPC)　and　PP　beta　layer　facing　MC　cells　(PPC-PP-PP　beta)　to　evaluate　surfacematerial　effects.　Membrane　properties　and　structures　were　evaluated.　Antibacterial　properties　against　Streptococcus　mutans　and　Staphylococcus　aureus　were　determined.　Scanning　electron　microscope　demonstrated　smaller　interfiber　spaces　of　PPC　and　PP　beta-PP-PPC　compared　to　PP　beta,　PPC-PP-PP　beta,　and　PP.　PPC　and　PP　beta-PP-PPC　exhibited　hydrophilic　property.　The　three-layer　membranes　(PPC-PP-PP　beta　and　PP　beta-PP-PPC)　demonstrated　significantly　higher　Youngs　modulus　(94.99　+/-　4.03　MPa　and　92.88　+/-　4.03　MPa)　compared　to　PP　(48.76　+/-　18.15　MPa)　or　PPC　(7.92　+/-　3.97　MPa)　(p　＜　0.05).　No　significant　difference　of　cell　proliferation　was　found　among　any　groups　at　any　time　point　(p　＞　0.05).　Higher　expression　of　integrins　were　detected　at　12　h　of　cultures　on　PPC-PP-PP　beta　compared　to　the　controls.　Promoted　osteoconductive　effects　of　PPC-PP-PP　beta　were　revealed　by　alkaline　phosphatase　assays　and　Western　blot　compared　with　the　controls　at　7　and　14　days.　PPC,　PPC-PP-PP　beta　and　PP　beta-PP-PPC　exhibited　a　significantly　wider　antibacterial　zone　against　the　tested　bacteria　compared　to　PP　and　PP　beta　(p　＜　0.05).　These　results　suggested　that　the　three-layer　electrospun　membranes　demonstrated　superior　properties:　higher　strength,　better　cell　adhesion,　and　promoted　osteoconductive　properties　compared　to　single-layer　membrane:　however,　antibacterial　properties　were　exhibited　in　three-layer　electrospun　membranes　and　chlorhexidine-doped　single-layer　membrane.　We　concluded　that　the　novel　three-layer　membranes　could　be　used　as　a　biocompatible　scaffold　for　intraoral　bone　regeneration　due　to　its　enhanced　osteoconductive　activity　and　antibacterial　effect.