Skin wounds may threaten quality of life and cause serious complications. This study aimed to investigate the effects of lyophilized exopolysaccharide (L-EPS) obtained from the probiotic strain Lactiplantibacillus plantarum GD2 on various stages of wound healing. The results revealed that L-EPS accelerated in vitro wound healing and increased COL1A1 in L929 cells. L-EPS affected the TGF-β1/Smad signaling pathway by increasing the expression of the TGF-β1, Smad2, Smad3, and Smad4 genes. L-EPS also exerted anti-inflammatory effects by reducing the gene expression of IL-1β, IL-6 and iNOS in TNF-α-induced fibroblasts. Additionally, L-EPS demonstrated fibroproliferative effect on both healthy and TNF-α-induced fibroblasts. Furthermore, L-EPS was found to have a proangiogenic effect in ovo chorioallantoic membrane (CAM) model. This study presents the first-ever characterization of the multifaceted effects of L-EPS derived from the probiotic strain L. plantarum GD2 on wound healing. Our findings highlight the potential of L-EPS as effective agent for wound healing and suggest possible application in the development of wound healing biomaterials. By elucidating the mechanism of action of L-EPS in wound healing, this research may provide new perspectives for advanced treatment strategies in the field of wound care.