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Ma H et al. Laser therapy for wound healing medium-fluence laser light (20–80 J/cm2) [11,13] tends to accelerate cell growth and wound-healing, whereas higher- fluence light (more than 80 J/cm2) [18] negates the beneficial effect of laser exposure. However, no significant difference or inhibited effect in fibroblast proliferation has also been found using lower-fluence laser light in several studies [8,10]. Therefore, lower-fluence laser light remaining controversial also would not be an option for our study. A similar conclu- sion has also been found by Luciana Almeida-Lopes [19]. As a result, the better choice of fluence for our study was to use one of medium-fluence laser light [4,12]. The fluence at 60 J/cm2 was used in our study. However, a dose-dependent study is needed in the future to definitely determine its effect on cell proliferation and collagen synthesis of fibroblasts. Our findings indicated that LLLT at wavelength of 830 nm or 635 nm+830 nm with fluence of 60 J/cm2 could modulate fibroblast proliferation and collagen synthesis, with cells irra- diated at 635 nm+830 nm possibly giving better results. It is difficult to compare our results with observations of other studies in the literature, because most studies have reported only one or two different single wavelength and treatment de- sign varies depending on individual studies [10,11]. However, some studies have been performed in a similar manner to our study [12,20,21]. Bogdan Crisan has compared near-in- frared laser light at wavelength of 830 nm, 980 nm, and 2,940 nm. Whereas, these three wavelength are actually is in the same spectrum. Among these three wavelength, irradia- tion at 830 nm showed the most notable bio-stimulation ef- fect on human skin fibroblasts [21]. For comparison between two kinds of spectrum (visible red light and infrared light), some studies have demonstrated that infrared low level lasers are preferred over visible red light lasers in stimulating cell proliferation or collagen synthesis [12,19]. These results are similar to results of the present study. It has been reported that 780 nm laser light can up-regulate the production of ba- sic fibroblastic growth factor (bFGF) more than 660 nm visi- ble red laser [12]. Diabetic fibroblasts irradiated at 830 nm have shown more bFGF release compared to those irradiated at 632.8 nm [20]. Previous studies have demonstrated action mechanisms of visible to near-infrared radiation on cells. It has been sug- gested that we should pay an attention to energy values rele- vant to the respiratory chain of cells. The effect of electro- magnetic wavelength on cellular energy transfer has become evident [16]. Tiina Karu reported that absorbing molecule can transfer energy to another molecule and that the activated molecule can then cause chemical reactions in surrounding tissue as the basis for LLLT effect [13]. Moreover, cyto- chromes c oxidase (red and near-infrared light region) is one of the primary photoacceptors reported in the literature. Co- factors for cytochromes c oxidase and cytochromes b, c1, and c are porphyrins while cofactors for complex I (an en- zyme of the respiratory chains) are flavins [22]. These changes in mitochondrial respiratory chain can mediate the activation or suppression of signal molecules in the cyto- plasm, causing subsequent changes of downstream cas- cades and leading to the synthesis of DNA, RNA, proteins, and enzymes in the nucleus, cytoplasm, or plasma mem- brane. Finally, these changes will result in photobiological ef- fects of cells such as cell proliferation and differentiation [23, 24]. Therefore, we speculated that dual-wavelength (635 nm+830 nm) could have better effects on fibroblast prolifera- tion and collagen synthesis through both red region and near- infrared region action spectrum by increasing the range of photoacceptor molecule. However, according the present study, we can’t demonstrate that there are significant differ- ence in cellular response of human fibroblasts between 830 nm and 635 nm+830 nm, may be due to the less amount of case. A further study is needed in the future to confirm it. The results of the present study may suggest that dual-wave- length light (635 nm+830 nm) could be an optimal choice for LLLT to improve the wound-healing. Our results might help clinicians in the choice of wavelength appropriate for LLLT. As a conclusion, our study demonstrated that dual-wave- length light (635 nm+830 nm) or infrared light (830 nm) has stimulating effect on proliferation and collagen synthesis of human fibroblasts in vitro. However, visible red light (635 nm) does not. Conflict of interest No potential conflict of interest relevant to this article was re- ported. Acknowledgment This article was presented as a oral presentation at the Inter- narional 75th Congress of the Korean Society of Plastic and Reconstructive Surgeons on Nov 10–12, 2017. https://doi.org/10.22467/jwmr.2018.00283 www.jwmr.org 5PDF Image | Low-Level Laser Therapy on Proliferation and Collagen Synthesis of Human Fibroblasts
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