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FIG. 5. LED response at 8 J/cm2; 50 mW/cm2 using individual wavelengths of 670, 728, and 880 nm (percentage change from control versus number of hours after LED treatment). seeded in 12-well plates with a well surface area of approxi- mately 4 cm2. DNA synthesis was determined on the second, third, and fourth days in culture for both fibroblasts (Fig. 1) and osteoblasts (Fig. 2). Exposure to LED irradiation accelerated the growth rate of fibroblasts and osteoblasts in culture for 2–3 days (growing phase), but showed no significant change in growth rate for cells in culture at 4 days (stationary phase). These data are important demonstrations of cell-to-cell contact inhibition, which occurs in vitro once cell cultures approach confluence. This is analogous, in vivo, to a healthy organism, which will regenerate healing tissue, but stop further growth when healing is complete. It is important to note that LED treat- ment accelerates normal healing and tissue regeneration with- out producing overgrowth or neoplastic transformation. A series of experiments has recently been completed using an L-6 musculoskeletal cell line (rat derived). These cells were exposed to the LED light at both combined wavelengths and in- dividual wavelengths (670, 728, and 880 nm), energy densities of 4 and 8 J/cm2, and an intensity of 50 mW/cm2. Results demonstrated a cell growth increase of about 140% over un- treated controls, particularly at 8 J/cm2 energy, as shown in Figure 3. In addition, experiments are now complete using a normal human epithelial cell line seeded in 12-well plates at a concen- tration of 500 cells/well in order to possibly explain the contin- ued, dramatic results of LED light therapy in preventing oral mucositis in cancer patients. Cell growth increased 155% over untreated controls at 670 nm and 4 J/cm2 energy density (50 mW/cm2 power density), as shown in Figure 4. An increase of 171% over untreated controls was obtained with a wavelength of 880 nm and 8 J/cm2 energy density (53 mW/cm2 power den- sity), as shown in Figure 5. Collagen synthesis of the HaCAT epithelial cells was deter- mined by measuring tritiated proline incorporation using a modified method described by Peterkofsky and Diegelmann.24 HaCAT epithelial cells were seeded in two 12-well tissue cul- ture plates with 600 mL DMEM containing 10% FBS, 1% peni- cillin/streptomycin, and 6 mL L-proline [2,3-3H]. The media was free of nonessential amino acids. One plate was used as the control, and the other was treated with the 670-nm NASA LED Whelan et al. 308 FIG. 6. HaCAT epithelial cell collagen synthesis at 8 J/cm2, 50 mW/cm2, 670 nm. Shown in 24-h 3H proline incorporation.PDF Image | Effect of NASA Light-Emitting Diode Irradiation on Wound Healing
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