Immunohistochemical identification of vascular endothelial growth factor in pig latissimus dorsi musculocutaneous flaps following ischemia-reperfusion injury

Vascular Endothelial Growth Factor (VEGF), a potent angiogenic, mitogenic and vascular permeability enhancing protein, appears to improve survival of ischemic flaps independent of its route of administration. The purpose of this study was to examine VEGF protein expression in biopsies of surgical flaps with immunohistochemical techniques. In 6 male Yorkshire-type pigs, 10 cm x 15 cm Latissimus dorsi musculocutaneous flaps were elevated bilaterally. Flap zones I, II, and III were established according to their distance from the vascular pedicle. After isolation of the thoracodorsal artery and vein, one flap was randomly assigned to ischemia by temporary occlusion of the vascular pedicle. Ischemia (4 hours) was followed by 2 hours of reperfusion (ischemia group, n=6). The contralateral (nonischemic) flap served as a control (control group, n=6). Skin and muscle biopsies of flaps were taken at the end of the protocol for immunohistochemical staining using a VEGF antihuman monoclonal antibody. Epidermis of flap skin did not demonstrate VEGF-positive staining, but the dermis and subcutaneous tissue did. Muscle components of biopsies demonstrated staining of interfascicular septa and staining of myocytes. A semi-quantitative scoring system with a scale of 0 to 3 was used for grading of immunohistochemical staining. In skin, areas adjacent to the flap showed an overall mean VEGF staining score of 0.7. All zones of ischemic flaps showed increased mean immunohistochemical staining for VEGF (scores=1.2, 1.6, and 1.4 in zones I, II, and III, respectively). In muscle, however, only zone I showed increased VEGF immunohistochemical staining from 0.7 in adjacent areas to 1.7 in ischemic flaps. The results indicate only moderate endogenous up-regulation of VEGF in flaps, supporting the utilization of exogenous VEGF as an adjunct in microsurgical therapy.