Sliver-based dressings have excellent antibacterial properties, but also show strong cytotoxicity. To solve this problem, a novel multifunctional sliver nanowires/collagen I/bacterial cellulose (AgNW/CBC) dressing was constructed via compositing silver nanowires (AgNWs) into bacterial cellulose (BC) structure using step-by-step in situ biosynthesis method, followed by adsorbing collagen I (Col-I). The results of SEM indicated that Col-I aggregated on the surface of BC under the action of electrostatic attraction between AgNWs and Col-I. the obtained AgNW/CBC dressings exhibited acceptable thermostability, favorable water absorbency, improved mechanical properties and water retention capacity and slower release of silver ions, compared with sliver nanowires/bacterial cellulose (AgNW/BC). In vitro results indicated that AgNW/CBC has excellent antibacterial activity against bacteria ATCC 25923 and ATCC 25922, as well as AgNW/BC, but also exhibit improved cytocompatibility to promote NIH3T3 cells proliferation and spread compared with BC and AgNW/BC. The results of the in vivo assay also confirmed that AgNW/CBC dressings can eliminate S. aureus infection and promote wound healing within 8 days, accompanied by inhibiting the inflammation, promoting collagen deposition, hair follicle growth and neovascularization. The AgNW/CBC dressings with favorable multifunctional properties are of considerable value to meet clinical requirements.