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NASA LED Irradiation and Wound Healing 309 FIG. 7. Change in wound size in rat ischemic wound model versus time (days). at 8 J/cm2. After 24 h of incubation in 5% CO2 at 37°C, two 200-mL aliquots of media were removed from each well. One aliquot was used to quantitate total protein by trichloroacetic acid (TCA) precipitation. The precipitated proteins were col- lected by suction onto a glass fiber filter and allowed to dry overnight at room temperature. The second aliquot was incu- bated for 90 min at 37°C with highly purified bacterial collage- nase that degraded the collagen in the sample. The remaining noncollagen protein also underwent TCA precipitation. The following day, scintillation fluid was added and the samples were counted in a scintillation counter. Collagen content was determined by subtracting the noncollagen protein from the to- tal protein. Figure 6 shows that the HaCAT epithelial cells that were NASA LED treated synthesized more than twice the amount of collagen than that of the control cells. LED WOUND HEALING IN RATS An ischemic wound is a wound in which there is a lack of oxygen to the wound bed due to an obstruction of arterial blood flow. Tissue ischemia is a significant cause of impaired wound healing, which renders the wound more susceptible to infection, leading to chronic, nonhealing wounds. Despite progress in wound healing research, there is still very little understanding of what constitutes a chronic wound, particularly at the molecu- lar level. Consequently, there is minimal scientific rationale for treatment. In order to study the effects of NASA LED technology and HBO therapy, we developed a model of an ischemic wound in normal Sprague-Dawley rats. Two parallel, 11-cm incisions were made 2.5 cm apart on the dorsum of the rats, leaving the cranial and caudal ends intact. The skin was elevated along the length of the flap, and two punch biopsies created the wounds in the center of the flap. A sheet of silicone was placed between the skin and the underlying muscle to act as a barrier to vascu- lar growth, thus increasing the ischemic insult to the wounds. The four groups, each consisting of 15 rats, in this study include the control (no LED or HBO), HBO only, LED (880 nm) only, and LED and HBO in combination. The HBO was supplied at 2.4 atm for 90 min, and the LED was delivered at a fluence of 4 J/cm2 and 50 mW/cm2 for 14 consecutive days. A future study FIG. 8. Change in vascular endothelial growth factor (VEGF) concentration (mg/mg protein) versus time (days).PDF Image | Effect of NASA Light-Emitting Diode Irradiation on Wound Healing
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