Introduction: Positive effects of topical application of honey on open skin wounds have been observed, Mast cell are playing role in normal wound healing and its disturbances but the role of mast cells in wound healing in existence of honey is uncertain. Objective: The aim of this investigation was to study the effects of two times per day topical application of honey on the number of mast cells in skin wounds. Materials and Methods: 33 male rats were randomly divided into experimental and control groups. Under general anesthesia and sterile conditions, one round full thickness skin wound was made on the dorsum of neck of each rat. The diameter of wounds was about 3cm. Unboiled honey was applied topically on wounds twice per day from surgery day until certain days. On the fourth, seventh, and fourteenth days after surgery according to inflammation, proliferation and remodeling phases of wound healing process, samples were obtained from wound tissue and adjacent skin after rats were killed with ether. Samples were prepared for routine histological study and stained with 1% aqueous toluidine blue. Mast cells of wounds were counted and analyzed by Mann Whitney U test. Results: The mean of mast cells was lower in experimental group in three above-mentioned days. The differences were statistically significant on days 4 and 7 (P<0.05 for fourth day, P<0.01 for seventh day, and P>0.05 for fourteenth day). Conclusion: Twice per day application of honey on open full thickness skin wounds could reduce the number of mast cells significantly, in inflammatory and proliferative phases of wound healing.