The 6 Gy of whole-body irradiation (WBI) with gamma rays results in an impairment of injured skin tissue recovery and renders a delay in the healing process. For an understanding of whether WBI has damaging effects on fibroblasts in wounds, fibroblasts in wounds combined with WBI and those of simple incision were isolated and cultivated, and abilities connected with tissue repair, including proliferation, attachment, adhesion, and apoptosis, were determined by direct cell count, immunohistochemical staining for proliferation cell nuclear antigen (PCNA), and TUNEL assay. The results showed that the abilities of proliferation and the attachment and adhesion of fibroblasts from wounds combined with WBI significantly decreased in comparison with those having simple incisions on the 3rd and 5th days of posttrauma, whereas the apoptotic ratio of fibroblasts from wounds combined with WBI significantly increased. These data suggest that WBI may exert damaging effects on fibroblasts in wounds, which might be one of the dominant reasons for the impaired healing of wounds combined with WBI.