Background: The process of wound healing is intricate and dynamic, characterized by the interactions of various cell types, e.g. fibroblast cells. Re-epithelialization phase encompasses a sequence of events involving the migration, proliferation, and differentiation of fibroblasts present at the wound site. Exopolysaccharides (EPS) are soluble and insoluble biopolymers, produced by a range of microorganisms including fungi, bacteria, microalgae, and yeast. The production of microbial EPS frequently escalates in response to environmental stressors, serving as a mechanism to cope with such challenges. These biopolymers are characterized by their high molecular weight, consisting of smaller sugar units known as monosaccharides that are linked together via glycosidic bonds. Due to their distinct structures and properties, microbial EPS have various scientific and medical applications, including regenerative medicine and wound healing. In the current research, we studied the effect of an EPS produced by a GRAS (generally regarded as safe) lactic acid bacterium, designated Limosilactobacillus sp. UTMC ۳۸۲۳ on the migration of Human Foreskin Fibroblast cells (HFF-۱) in vitro. Methods: The strain Limosilactobacillus sp. UTMC ۳۸۲۳ was cultivated in MRS broth supplemented with ۱۰% sucrose at pH ۵.۷, ۳۷°C, ۴۸ h for EPS production. After removing the biomass through centrifugation at ۲۵۰۰ g, ۲۰ min, the supernatant was mixed with equal volume of ethanol at ۴°C. The precipitate was separated and considered as crude EPS. Further purification was done by trichloroacetic acid, and the purified EPS was freeze-dried and kept in - ۷۶ o C before use. MTT test was performed to evaluate the toxicity of ۰-۵ mg/ml concentrations of EPS against human foreskin fibroblasts (HFFs) cells. Also, the effect of EPS on cell migration was evaluated by scratch wound assay. The HFF fibroblast cells were seeded in ۲۴ wells plates and were allowed to proliferate and adhere to the plate overnight. Then, vertical wounds were created at the cell surface by a sterile ۱۰ μl pipette tip. The EPS was added to cells and incubated at ۳۷ °C for ۴۸ h.
