Departments of Otolaryngology-Head and Neck Surgery, University of Minnesota Medical Center, Minneapolis, Minnesota 55455, USA.
Otolaryngol Head Neck Surg. 2013 Sep;149(3):384-9. doi: 10.1177/0194599813492644. Epub 2013 Jun 6.
To evaluate clinical and immunohistopathological effects of topical glycyl-histidyl-lysine-copper (GHK-Cu) on in vivo irradiated rat wounds.
Animal model.
Academic institution.
After dorsal irradiation and a 28-day recovery period, 2 × 8 cm cranially based dorsal flaps were created in Sprague-Dawley rats. Twice daily GHK-Cu gel (test) or aquaphilic ointment (control) was applied for 10 days. Animals were euthanized, digital images of flaps were taken, and harvested tissues were immunohistochemically stained for a vascular endothelium marker, caveolin-1, and vascular endothelial growth factor (VEGF). Digital image analysis was used for outcome measures. Unpaired t-tests were used for statistical analyses; significance of P < .01 accounted for multiple comparisons.
By digital analysis of clinical images, 13 test and 10 control animals showed mean ischemic areas of 5.0 cm(2) (SD = 0.9) for tests and 3.8 cm(2) (SD = 1.1; P = .011) for controls. Whole slide digitized images allowed quantification of caveolin-1-stained blood vessels and VEGF expression in fibroblasts at the interface of healing flaps. Caveolin-1 analyses showed a mean of 209.0 vessels (SD = 111.1) and a mean vessel luminal area of 525.7 um(2) (SD = 191.0) in tests and 207.4 vessels (SD = 109.4; P = .973) and 422.8 um(2) (SD = 109.7; P = .118) in controls. VEGF quantified as the percentage of pixels exceeding a colorimetric threshold, with higher fractions of positive pixels indicating more intense staining, showed a mean intensity score of 0.34 (SD = 0.19) in tests and 0.54 (SD = 0.41; P = .169) in controls.
Irradiated dorsal rat flaps treated with topical GHK-Cu gel demonstrated no difference in flap ischemia, blood vessel number or area, or VEGF expression compared to controls.
评估局部甘氨酰-组氨酰-赖氨酸-铜（GHK-Cu）对体内放射大鼠伤口的临床和免疫组织病理学影响。
动物模型。
学术机构。
背部照射后和 28 天恢复期后，在 Sprague-Dawley 大鼠背部创建 2×8cm 颅基背皮瓣。每天两次应用 GHK-Cu 凝胶（试验）或亲水软膏（对照）治疗 10 天。处死动物，拍摄皮瓣的数字图像，并对采集的组织进行血管内皮标志物、窖蛋白-1 和血管内皮生长因子（VEGF）的免疫组织化学染色。使用数字图像分析进行结果测量。使用未配对 t 检验进行统计分析；P<0.01 表示多重比较的显著性。
通过对临床图像的数字分析，13 只试验和 10 只对照动物的试验组平均缺血面积为 5.0cm²（SD=0.9），对照组为 3.8cm²（SD=1.1；P=0.011）。全幻灯片数字化图像允许定量分析愈合皮瓣界面处窖蛋白-1 染色的血管和成纤维细胞中的 VEGF 表达。窖蛋白-1 分析显示，试验组的平均血管数为 209.0 个（SD=111.1），平均血管腔面积为 525.7μm²（SD=191.0），对照组分别为 207.4 个（SD=109.4；P=0.973）和 422.8μm²（SD=109.7；P=0.118）。VEGF 作为超过比色阈值的像素百分比进行量化，较高的阳性像素分数表示更强的染色强度，试验组的平均强度评分为 0.34（SD=0.19），对照组为 0.54（SD=0.41；P=0.169）。
与对照组相比，局部应用 GHK-Cu 凝胶治疗的辐射性大鼠背部皮瓣在皮瓣缺血、血管数量或面积或 VEGF 表达方面没有差异。
