In this study, a kind of antibacterial collagen (Coll)/cellulose nanocrystals (CNCs) scaffolds was prepared and characterized, which can be applied in skin trauma as a promising dressing. In order to achieve the goal of antibacterial function for a period of long time, the Gentamycin sulfate (GS) was absorbed in the prepared gelatin microsphere (GMs) then doped in the porous Coll-CNCs freeze-drying scaffold. The CNCs had a width of 20-40 nm and length of 150-350 nm from electron microscopy (TEM), retained the basic chemical structure of cellulose observed from Fourier transform infrared, and increased the machinery properties of Coll scaffold from thermal property analysis. From the SEM, the mean pore size of the compound scaffolds was about 120 mu m with or without GMs. The in vitro release profile of GS showed a sustained release of this GS/GMs/Coll-CNCs composite scaffold. The biological compatibility experiments showed the composite scaffold low cellular cytotoxicity of NIH-3T3 cells. The significant inhibition zones indicated the antibacterial efficacy of the composite scaffold against Escherichia coli and Staphylococcus aureus.