Two-dimensional (2D) nanopore membranes play a pivotal role in the application of molecular separation, DNA sequencing, drug release and biosensing. Owing to their special properties including 2D symmetry and interfacial activity, amphiphilic nanosheets (ANs) have great potential for generating functional nanomaterials. In this work, the dissipative particle dynamics (DPD) simulations are employed to design 2D nanopore membranes with tunable size using AN molecules. We find that these two-dimensional membranes are much stable due to the double-layer assembled framework of the membrane with the thickness from 2.5 to 7.8 nm. Different sized nanopores ranging from 2.45 to 6.15 nm are also identified in the membrane architecture, which can be controlled by the concentration ratio, the length of grafting polymer and the affinity of ANs. Moreover, when at low concentration ratio (<10%), the ANs tend to form different core-shell structured micelles, including spherical, ellipsoidal and long rod-like self-assembled morphologies. From our study findings, the ANs could be extendedly used for drug delivery and release, multi-layer 2D nanomaterials by the advantage of their chemical tunability, including the affinity and stimulus-responsive properties of grafting polymer as well as the geometry of scaffold structure in the ANs. (C) 2018 Elsevier Ltd. All rights reserved.