光刺激對纖維母細胞及角質細胞的影響

Effect of light irradiation on human foreskin fibroblast and keratinocyte

謝岳峰(Yue-Feng Hsieh)

指導教授 ： 阮若屈 ； 謝瑞香

中原大學/工學院/醫學工程研究所/碩士(2005年)

https://doi.org/10.6840/cycu200500692

摘要

低能量雷射用於傷口癒合的治療已經有三、四十年之久，但結果不盡相同，且低能量雷射的操作不方便，可用波長範圍狹小，照射面積太小，操作不方便及價格昂貴等問題，故以發光二極體(LED)做刺激光源，探討低能量雷射對於傷口癒合的影響。
實驗探討在傷口癒合中兩種有關係的細胞：纖維母細胞及角質細胞。為了完全了解光刺激的影響，利用發光二極體探討各種可控制參數，包括不同的細胞、波長(465、630 nm)、功率密度(1.5~16.3 mW/cm2)、光照時間(100~800 sec)及總能量(0.3~6.8 J/cm2)對纖維母細胞及角質細胞的影響。同時也會探討光刺激對細胞分泌細胞激素(TGF-β1、KGF)及膠原蛋白的影響。
研究結果發現，以紅光630 nm可促進纖維母細胞的增生，而藍光則否；而隨著功率密度、光照時間及總能量的增加，纖維母細胞增生速率也隨之增加；不過在相近的能量下，似乎是功率密度對纖維母細胞的增生有較大的影響。而較佳的刺激纖維母細胞增生參數為總能量6.52 J/cm2 (16.3 mW/cm2 x 400sec)。
以此參數做生長因子探討，發現光刺激後的半小時後，都有明顯促進纖維母細胞分泌更多的transforming growth factor β1(TGF-β1)及Keratinocyte growth factor (KGF)，但卻不會促進纖維母細胞分泌更多的膠原蛋白；另外在光刺激角質細胞實驗中發現，發光二極體似乎不會刺激角質細胞的增生。而光刺激後纖維母細胞增加分泌TGF-β1和KGF或許暗指可加速傷口的癒合。由以上結果顯示，在630 nm紅光下，以功率密度16.3 mW/cm2，光照時間為400秒，即在總能量為6.52 J/cm2情況下，可刺激纖維母細胞增生，因此紅光630 nm或可能加速傷口癒合速度。

關鍵字

纖維母細胞、角質細胞、發光二極體、轉形生長因子、角質細胞生長因子、膠原蛋白

並列摘要

Low-level laser applied on wound healing had been for 30 to 40 years, but the benefits of low-level laser in wound healing are still controversial. The limited selectable wavelength and the bulky size restricted the extensive studies of various operating variables.
In this study, we investigated the effect of light stimulation on two of the wound healing related cells, human foreskin fibroblasts and keratinocytes. In order to thoroughly understand the effects of light, we used different LED light panels to simultaneously investigate all the controllable parameters, which include wavelength and power density of stimulating light, treating time per exposure, as well as the total applied energy. We compared cell proliferation when stimulated at the similar total applied energy under different combinations of light intensity and stimulating time which one is more important to proliferate cell growth.
We also demonstrate the effect of light irradiation by the secretion of TGF-β1、KGF and collagen.
The results showed that single dose of light treatment was sufficient to accelerate fibroblast proliferation when cells were exposed under red light (630 nm) at a power density of 8.5 mW/cm2 but the blue LED light had no effect on cell growth. Compared to the non-irradiated control, the acceleration of cell proliferation increased with the increase in irradiation time、light intensity and total applied energy. We also found light intensity was found more effective in growth stimulation comparing to irradiation time under the similar total applied energy. Our experimental result was found the better of irradiation parameter was 6.52 J/cm2 (16.3 mW/cm2 x 400sec)
The secretion of TGF-β1, KGF and collagen of stimulated cells was also under investigation. We did not find any measurable difference in collagen secretion after light irradiation, but the secretion of TGF-β1 and KGF increased significantly at half an hour after each treatment. But there was no effect between irradiation and non-irradiation on keratinocytes. The increase in TGF-β1 and KGF secreted from fibroblasts indicated that it might be possible to enhance wound healing through light treatment.

並列關鍵字

Fibroblast、Keratinocyte、LED、TGF-β1、KGF、Collagen

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