Polydeoxyribonucleotide (PDRN) has multiple vascular actions such as angiogenesis and production of a vascular endothelial growth factor (VEGF) through the adenosine A2 receptor stimulation. We applied PDRN on the ischemic flap of rat back and investigated whether it enhances the skin flap survival. A total of 28 Sprague-Dawley rats were divided into 3 groups, namely, PDRN group, control group 1 (no treatment), and group 2 (phosphate-buffered saline injection). On the distally based flap of 3 × 9 cm in size, it was subdermally injected with PDRN or phosphate-buffered saline, which were administered 48 hours prior and immediately after flap elevation. The PDRN group was daily maintained by intraperitoneal administration of PDRN from the postoperative 1st day to 10th day. The mean survival rates of flap in PDRN group [79.5% (6.3%)] are significantly larger than control groups [1, 53.0% (6.9%); 2, 51.7% (6.7%)]. Serial measurements of blood perfusion also showed that the blood flux was significantly increased in almost part of the flap on the 10 days after PDRN injection. The number of CD31 positively stained vessels and expression of VEGF protein were significantly higher in the PDRN group. We propose that administration of PDRN into the ischemic skin flaps increased blood flux to the flap, VEGF expression, and number of capillaries, thereby improving the rat skin flap survival.