Polysaccharides could be used as biodegradable and biocompatible polymers for scaffolds and carriers matrix. Numerous algal, fungi and herbal polysaccharides can attenuate degradation of skin matrix by the inhibition of elastase, collagenase and matrix metalloproteinases (MMPs). In this study, we investigate anti-elastase and anti-collagenase potential of Lactobacilli exopolysaccharides (LEPS) on normal human fibroblast. Among 60 Lactobacilli isolated from herbal plants and dairy products, selected LEPS showed high anti-collagenase (up to 100%), anti-elastase (up to 87%) and antioxidant activity (up to 60%). Most of them had no cytotoxicity effect on fibroblast, and some of them promote cell proliferation (up to 10%). In scratch assay, all the investigated EPS stimulated wound healing process in fibroblast (up to 99%). MMP1, MMP2, MMP3, MMP9 and MMP10 were down-regulated significantly and TIMP1 and TIMP2 were up-regulated slightly in LEPS of B9-1 from L. casei with high anti-collagenase and anti-elastase activity; however, no meaningful alteration was observed in MMP expression level for LEPS of P35 from L. plantarum with low anti-collagenase and anti-elastase activity. By consideration of high anti-collagenase, anti-elastase, antioxidant activity and wound healing of LEPS, they could be considered as good candidate of skin anti-aging agents for tissue engineering and skin regeneration scaffolds.