Introduction: Although clinical studies have identified scarring/fibrosis as significant risk factors for lymphedema, the mechanisms by which lymphatic repair is impaired remain unknown. TGF-1 is a critical regulator of tissue fibrosis/scarring and may therefore also play a role in the regulation of lymphatic regeneration. The purpose of this study was therefore to assess the role of TGF-1 on scarring/fibrosis and lymphatic regeneration in a mouse tail model. Methods: Acute lymphedema was induced in mouse tails by full thickness skin excision and lymphatic ligation. TGF-1 expression and scarring were modulated by repairing wounds with or without topical collagen gel. Lymphatic function and histologic analyses were performed at various time points. Finally, the effects of TGF-1 on lymphatic endothelial cell (LEC) in vitro were evaluated. Results: Wound repair with collagen gel significantly reduced the expression of TGF-1, decreased scarring/fibrosis, and significantly accelerated lymphatic regeneration. The addition of recombinant TGF-1 to the collagen gel negated these effects. Improved lymphatic regeneration secondary to TGF-1 inhibition was associated with increased infiltration and proliferation of LECs and macrophages. TGF-1 caused a dose-dependent significant decrease in cellular proliferation and tubule formation of isolated LECs without changes in the expression of VEGF-C/D. Finally, increased expression of TGF-1 during wound repair resulted in lymphatic fibrosis and co-expression of -SMA and LYVE-1 in regenerated lymphatics. Conclusions: Inhibition of TGF-1 expression significantly accelerates lymphatic regeneration during wound healing. Increased TGF-1 expression inhibits LEC proliferation and function, and promotes lymphatic fibrosis. These findings imply that clinical interventions that diminish TGF-1 expression may be useful in promoting more rapid lymphatic regeneration.