Biomimetic materials are of great significance in wide applications. As a bioactive polysaccharide, chitosan (CS) has attracted much attention in multiple fields. Inspired by the natural shark skin interface, herein, we proposed a methodology to obtain serial biomimetic CS/glycerol membranes via a facile mold casting and solvent evaporation process under mild aqueous conditions. The surface morphology, chemical structures, and physical characteristics of the developed membranes were determined by scanning electron microscopy, Fourier transformed infrared spectroscopy, tensile measurement, water vapor transmission rate, liquid uptake behavior, solubility, and dynamic water contact angle tests. Furthermore, in vitro and in vivo biological assays were performed, including antibacterial ability, hemostatic performance, hemolytic reaction, cytotoxicity evaluation, and contaminated wound healing assessment. Compared with the flat membranes, the micro-patterned CS/glycerol membranes show the higher water-absorbing ability (up to ca. 26-fold@30 min), moderate tensile strength (8.5–14.7 MPa), superior bacteriostasis (84.9% to 98.6% against S. aureus, 84.3% to 99.6% against E. coli), decent hemostat efficacy (within 75.7 s), low hemolysis rates (less than 0.5%), and benign cytotoxicity (84.2% to 98.4% of viability), which favor promoting contaminated wound healing. Thus, the biomimetic micro-patterned membranes improve bioactive performance and show the merits of simple operation, low cost, and easy scale-up. All these properties indicate that the hybrid micro-patterned membrane could be used as a suitable candidate for dermal wound dressings and open up the insights for developing other bionic interfaces in future potential applications.