The aim of this study was developed by a simple method including solution mixing of cellulose acetate/sodium alginate containing laponite (CA/SA-LAP) hydrogel composites with flexible mechanical, physical, and biological properties for wound healing application. The surface properties of the as-prepared CA/SA hydrogel and CA/SA-LAP hydrogel composites were characterized by Fourier transform infrared spectroscopy (FT-IR) and X-Ray Diffraction (XRD). The influence of LAP content (0.1, 0.2, 0.5 wt%), the swelling ratio (171.0 ± 7.0, 143.0 ± 4.0, and 96.6 ± 2.5), degradation rate (65.7 ± 4.0%, 49.0 ± 5.5%, and 19.4 ± 1.4%), mechanical properties elongation at break from 68.3 ± 6.0% to 148.3 ± 5.6%, and the morphology of the CA/SA-LAP was investigated. By increasing the LAP concentration with an average pore size diameter decreased from 300  μm to 225  μm, 150  μm, and 75 μm. The as-prepared CA/SA-LAP dressing has designated good antimicrobial activity against Staphylococcus aureus and Escherichia coli bacteria at 95.9 ± 4.0% and 98.4 ± 1.5% for 24 h and 92.3 ± 4.5% and 96.4 ± 3.5% for 48 h. Furthermore, CA/SA-LAP revealed admirable biocompatibility against human osteosarcoma cell line MG-63. Noticeably, the MTT assay demonstrated that fibroblast proliferation significantly enhanced on 0.5 wt.% LAP in CA/SA-LAP compared to CS/SA hydrogel at 92.6 ± 4.2% and 96.4 ± 3.5% for 24 and 48 h. Systematic in vivo research of the CA/SA-LAP was conducted in the rat bone defect model. The in vivo results proved that the CA/SA-loaded LAP significantly promoted bone healing in rat defects, compared to the CA/SA hydrogels. These results demonstrated the great potential of CA/SA-loaded LAP in wound healing material in nursing care application.