COMP-angiopoietin 1 Gene Transfer Enhances Cutaneous Wound Healing: MATERIALS AND METHODS
Adenoviral vectors
Recombinant adenoviruses that expressed COMP- Ang 1 or LacZ were constructed using the pAdEasy vector system (Ade-COMP-Ang 1 and Ade-LacZ) (Ade-COMP-Ang 1 and Ade-LacZ were generous gifts from Prof. Gou Young Koh at Korea Advanced Institute of Science and Technology, Daejeon, Korea).
Animals and adeno viral treatment
Eighteen male Sprague-Dawley rats (200 ~ 250 g in weight) were housed two per cage, maintained under controlled environmental conditions (12 h light to dark cycle, temperature approximately 23oC) and provided with standard laboratory food and water ad libitum. All the animal procedures were in accordance with the declaration of Helsinki and the guidelines for the care and use of laboratory animals. The animal care and experimental procedures were performed under approval from the Animal Care Committees of the Catholic University of Korea. For the adeno-viral treatment, 109 p.f.u. of Ade-COMP-Ang 1 and Ade-LacZ diluted in 50 p L of sterile 0.9% NaCl was injected intravenously into nine rats, through the tail vein two days before surgery. buy kamagra
Animal wound model
General anesthesia with ketamine (80 mg/kg body weight)/xylazine (10 mg/kg body weight) was achieved by intraperitoneal injection. The hair on the back of each rat was shaved and the back skin was subsequently wiped with 70% ethanol. Three full-thickness wounds (1 cm in diameter, 2~3 cm apart to minimize the tension to adjacent wounds) were made on the back of each rat by excising the skin. The wounds were allowed to form scabs.
Determination of wound healing rate
To compare the rate of wound healing between the Ade-COMP-Ang 1- and Ade-LacZ-treated groups, the animals were photographed on days 0, 3, 7 and 14 after surgery. In addition, the wounds were copied exactly onto transparent film with an indicated standard unit. The photographs and transparent films were digitally processed and the areas of the wounds were calculated using the KS300 system (Zeiss, Jena, Germany). For each sample, the rate of the healing process was measured as a percentage of the area of the wound at each time-point compared to the area of the wound at day 0. Viagra Professional
