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KMID : 1377020150120010037
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Tissue Engineering and Regenerative Medicine
2015 Volume.12 No. 1 p.37 ~ p.42
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Effect of light-emitting-diode irradiation on the proliferation and migration in human gingival fibroblasts
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Park Jong-Tae
Hong Ki-Seok
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Abstract
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Purpose: The aim of this study is to evaluate proliferation, migration, and various gene expressions including insulin-like growth factor 1 (IGF-1), SMAD3, and collagen type I (COL1A1) during light-emitting diode (LED) irradiation of human gingival fibroblasts (HGF).
Materials and methods: HGF was seeded on the culture dish and stimulated using white (1.26 J/cm2) and red (0.3 J/cm2) wavelength LED for 9 min during 3 days. The distance between LED light and HGF was about 30 mm. To evaluate cell proliferation, MTT assay was carried out for 3 days after LED irradiation. After culturing for 1 and 3 days, cells were harvested and total RNA was isolated. To assess the expression of IGF-1, IGF-2, IGF-1R, SMAD3, COL1A1, and GAPDH, cDNA was synthesized and RT-PCR was performed. Whether mixed LED irradiation affect to intracellular adhesion molecule-1 (ICAM-1) mRNA expression of HGF, real time PCR was performed. To measure wound healing after LED irradiation, straight scratch was made and observed migrated cells to the scratched region using microscope. Transwell migration assay was carried out to measure migrated cells after LED irradiation. Migrated cells were stained using hematoxilin.
Results: Cell proliferation was significantly increased both 1 and 3 days after LED irradiation. IGF-1 and COL1A1 mRNA expression was increased in LED irradiated cells compared to control cells. However, IGF-1R, IGF-2, and SMAD3 expression was decreased in LED irradiated cells compare to control cells. In LED irradiated cells, ICAM-1 expression was decreased than control. Interestingly, LED irradiation dramatically promoted wound healing and cell migration compared to control cells. Especially after 20 h, most of the substrate was covered by migrated HGF after LED irradiation.
Conclusion: These results suggest that simultaneous use of red and white wavelength LED promotes proliferation and migration of human gingival fibroblasts in vitro.
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KEYWORD
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LED(light-emitting diode ), Cell proliferation , human gingival fibroblasts, cell migration, wound healing
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