Red alga Gracilaria species have been reported various to exhibit significant pharmacological activities. In this study, sulfated galactans (SG) from G. fisheri was investigated for its potential activity in wound healing using in vitro approach. L929 fibroblast cells were treated with different concentrations of SG and wound healing activity and collagen content determined. Expression of the specific mRNA transcripts and translated proteins was assessed. In a scratch wound healing culture, SG enhanced the rate of migration of the fibroblast cells, accompanied by increased expression of integrin β1, p-FAK, p-NF-κB, Ecadherin proteins, and the active form of MMP-2. In addition, SG increased expression of COL1A1 mRNA and protein and stimulated collagen production. In conclusion, this study demonstrates that SG from G. fisheri promotes would healing of fibroblasts, in part, through integrin-FAK and NF-κB-MMP signaling pathways. This suggests that SG could be a promising compound for further investigation of its potential on wound treatment.