Flightless I (Flii) is a member of the gelsolin family of actin remodelling cytoskeletal proteins. Our previous studies have shown that Flii is a crucial mediator of wound healing, playing important roles in cell adhesion, proliferation, migration and contraction with possible important functions in scar formation. Whisker follicles (vibrissae) of rats and mice have remarkable regenerative abilities. Following microsurgical removal of up to one third of the active lower follicle, these animals are capable of regrowing all of the amputated components that are essential for normal hair production. This phenomenon has also been observed experimentally in human hairs. Using the hair follicle regeneration model, we have attempted to identify the involvement of Flii in the scar-less regeneration of hair follicles. The bulbar region of vibrissae from rats as well as 3 strains of mice expressing various levels of Flii (low Flii+/-, normal Flii+/+ and high Flii+/+;FliiTg/Tg) were surgically amputated, then allowed to regenerate in vivo. Macroscopic and histological assessment of the regeneration process was compared between mice expressing different levels of Flii. Immunohistochemistry was also performed to determine the patterns of expression of Flii and other wound healing associated factors during hair follicle regeneration. Understanding the mechanism of scar-free regeneration in hair follicles has wider implications for reducing scar formation in other types of wounds, and this study investigates the role Flii plays in this process.