Purpose This study is aiming to fabricate a small diameter vascular engineering scaffold thatcombined pNSR32 (recombinant spider silk protein), PCL (polycaprolactone) and Gt (gelatin),to investigate the physicochemical properties and cytocompatibility of the tubular scaffold.Method The tubular scaffolds were fabricated by electrospinning combination of pNSR32, PCLand Gt with weight ratio of 5:85:10. The morphologies and properties of the scaffolds werecharacterized by scanning electronic microscope(SEM), contact angle and water uptake test.Cytocompatibility of the scaffolds, followed by co-culture with SDRAECs (Sprague Dawley RatAortic Endothelial Cells) were evaluated through SEM, MTT, Flow cytometer analysis, frozensection and immunofluoresence staining.Result pNSR32/PCL/Gt tubular scaffold (inner diameter =3 mm) showed high porosity of 86.2±2.9％, pore size of 2423±979 nm and average fibre diameter of 166±85 nm. Water uptake andcontact angle of the scaffolds reached 112.0±4.4％ and 45.7±13.7° respectively. SDRAECsgrew and proliferated best on the pNSR32/PCL/Gt scaffolds followed by pNSR32/PCL,and finallyPCL. Particularly, not only the typical cobblestone morphology of confluent endothelial cellmonolayer was visualized but also factor VIII was normally expressed on pNSR32/PCL/Gtscaffolds after SDRAECs were cultured on the pNSR32/PCL/Gt scaffolds for 7 days.Conclusion The pNSR32/PCL/Gt composite tubular scaffolds presented appropriatecharacteristics and cytocompatibility would be considered as candidate scaffolds for vasculartissue engineering.