Latanoprost is an analogue of prostaglandin F(2alpha) which is used to treat glaucoma. Increases in eyelash number, thickness, and pigmentation have been reported as latanoprost side effects. The aim of this study was to evaluate if topical use of this drug can be used as a treatment of alopecia areata of eyebrows and eyelashes or not. In an experimental study, 26 patients with symmetrical eyelash and eyebrow alopecia areata were treated over 4 months with topical latanoprost for one side and the other side was not treated with any drug. The results were compared. Only one of the latanoprost-treated cases showed partial hair regrowth on the treated side. The relationship between hair regrowth and latanoprost application was not statistically significant (P = 1) by Fisher test. Based on our results, topical latanoprost has no efficacy in the treatment of alopecia areata. More studies with a larger sample size, longer study duration, and higher concentration of medication are suggested.