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研究生：	劉鳳起 Liou, Feng-Chi
論文名稱：	近紅外光有機發光二極體光照對於糖尿病大鼠傷口癒合之影響 Effect of Near-Infrared Light Irradiated by OLED on Wound Healing of Diabetic Rat
指導教授：	陳家進 Chen, Jia-Jin
學位類別：	碩士 Master
系所名稱：	工學院 - 生物醫學工程學系 Department of BioMedical Engineering
論文出版年：	2012
畢業學年度：	100
語文別：	英文
論文頁數：	42
中文關鍵詞：	糖尿病大鼠、傷口癒合、有機發光二極體、光治療、近紅外光譜、拉伸測試
外文關鍵詞：	Diabetic rat, Wound healing, OLED, Phototherapy, NIRS, Tensile test
相關次數：	點閱：101 下載：0
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目前已有足夠的證據顯示光治療可促進傷口癒合，且幾乎無副作用。而本研究之目的是利用有機發光二極體(OLED)治療糖尿病大鼠傷口模型，從中評估其光療的療效。三十隻糖尿病大鼠隨機分為治療組和控制組，每隻大鼠身上左右兩側各開一個方型傷口，僅治療組左側傷口經由632nm波長的OLED光照，其能量密度4 J/cm2，功率密度3 mW/cm2，每二天給予一次光治療，其餘的傷口則不接受治療。為了評估治療效果，本研究使用影像軟體ImageJ計算傷口面積的大小，近紅外光譜(NIRS)量測傷口或期邊緣區域的血紅素濃度，蘇木精與伊(H&E)染色評估傷口的組織結構，以及傷口癒合後使用拉伸測試評估皮膚組織之張力。本研究發現隨著傷口逐漸癒合，治療組(有照光)之傷口收縮百分比相較於控制組有顯著(p < 0.001)的差異。而在傷口癒合的增生期，NIRS量測的數據顯示控制組傷口區域的血紅素濃度高於治療組。本研究也發現在手術後的2個月(傷口皆已癒合，此時間點為傷口的重建期)，治療組傷口組織拉伸最大之張力約微控制組的2.6倍。H&E染色則顯示術後第10、22天的控制組傷口，相對於治療組有較嚴重的白血球浸潤。總結來說，OLED光療可加快傷口的癒合，降低發炎反應，以及增強癒合後的皮膚張力。

Sufficient evidence suggests that phototherapy can improve wound healing of diabetes with no side effects. The purpose of this study is to evaluate the effect of organic light-emitting diode (OLED) phototherapy in diabetic rat model. Thirty diabetic rats were used and divided into treatment and control groups. In treated rats, the left-flank wound was irradiated with 610 ~ 670 nm OLED wavelength peak at 632 nm, 4 J/cm2, 3 mW/cm2, 22.22 minutes every two days. The right flank of treatment group and both sides of controls were not irradiated. To assess the treatment effects, the wound area was evaluated by image processing software (ImageJ) as well as from the hemoglobin concentration measurement on the wound surrounding area using near infrared spectroscopy (NIRS). Furthermore, the tissue structure was evaluated using hematoxylin and eosin (H&E) staining and the skin tension of wound closure was measured using tensile tests. Our study indicated that significant differences (p < 0.01) between treatment-irradiated and control group for percentage of wound contraction during the wound healing. The NIRS results showed that the hemoglobin concentration on the wound of control was higher than that of treatment group at the proliferation phase. The high hemoglobin concentration might be related to the protracted inflammatory response of diabetic wound. Skin tensile strength of treatment group was 2.6 times higher (p < 0.05) than that of control at two months after surgery. The H&E staining showed that the leukocyte infiltration of control was more serious than treatment group on 10 and 22 days post surgery. In summary, the OLED phototherapy can accelerate the healing rate, reduce the inflammatory reaction, and improve the skin tensile strength.

Contents
摘要	I
Abstract	II
謝誌	III
Content	IV
List of Figures	V
Chapter 1 Introduction	1
1.1 The process of wound healing	1
1.2 The wound under influences of diabetes	3
1.3 Phototherapy	5
1.4 Principle of near infrared spectroscopy	7
1.5 Motivations and the aims of this study	8
Chapter 2 Materials and Methods 	10
2.1 Diabetic animal model	10
2.2 Phototherapy protocols using OLED	11
2.3 Wound assessment of area and hemoglobin	14
2.4 Histology	15
2.5 Skin tensile test	16
2.6 Statistical analyses	17
Chapter 3 Results 	18
3.1 Blood glucose and body weight monitoring in diabetics rat	19
3.2 Time-course change of wound area	20
3.3 The changes of hemoglobin during wound healing	25
3.4 Histological analysis	28
3.5 Mechanical tensile strength	31
Chapter 4 Discussion and Conclusion	34
References	37
List of Figures
Table 2.1 The changes of temperature during OLED irradiation measured by resistance temperature detector (room temperature is 24.3 ℃).	13
List of Figures
Figure 1.1 The time course of the different cells appearing in the wound during the healing process, and three overlapping phases of wound healing: inflammation, proliferation, and maturation/remodeling (Witte MB et al., 1997).	1
Figure 1.2 The processes of wound healing in detail (Beanes et al., 2003).	3
Figure 1.3 The mechanism of LLLT, ATP: Adenosine triphosphate. cAMP: Cyclic adenosine monophosphate. NO: Nitric oxide. ROS:Reactive-oxygen species (Huang et al., 2009).	6
Figure 1.4 Absorption factors of chromophores in “Optical Window”. Deoxyhemoglobin (blue line), oxyhemoglobin (red line), and water (black line) (Strangman et al., 2002).	8
Figure 2.1 Experimental design of animal model. Wound on the dorsum after one- month diabetic induction.	10
Figure 2.2 Wound modal design for four conditions (treatment-irradiated, treatment- unirradiated, control-left and control-right).	11
Figure 2.3 The spectrum of OLED light-emission.	12
Figure 2.4 (a) OLED device, (b) the treatment procedure, (c) the constant current driver circuit.	13
Figure 2.5 NIRS probe placement locations in animal model, the center of wound, the wound edge, and the control site (2 cm away from the wound edge).	15
Figure 2.6 Specification of specimens and mechanical testing system.	17
Figure 3.1 The physiological monitoring in diabetic rats: (a) body weight, (b) blood glucose level. Diabetic rats were induced on 35 days before wound surgery (day 0).	20
Figure 3.2 The percentage of wound contraction for each time point: (a) contro-left vs. control-right, (b) treatment-irradiated vs. treatment-unirradiated, (c) treatment- irradiated vs. control-left, (d) treatment-unirradiated vs. control-right. Asterisk represents the significant difference (p<0.05) between two groups by using independent t test.	23
Figure 3.3 The percentage of wound contraction for each time point. Asterisk represents the significant difference between three groups by using one-way ANOVA; * p<0.05, ** p<0.01.	24
Figure 3.4 The numbers of wound closure for four groups on each time point.	24
Figure 3.5 NIRS measurement of HbO concentration at wound edge, wound center and control site in four conditions: (a) treatment-irradiated, (b) treatment-unirradiated, (c) control-left and (d) control-right.	25
Figure 3.6 The Hb concentration measured by NIRS in four conditions: (a) treatment- irradiated, (b) treatment-unirradiated, (c) control-left and (d) control-right.	26
Figure 3.7 NIRS measurement of (a) HbO and (b) Hb concentration at wound edge. Asterisk represents the statistical difference (p<0.05) between two groups by using one-way ANOVA. Dashed line represents the mean of hemoglobin concentration at control site.	27
Figure 3.8 H&E staining of wound biopsies on 6 days after surgery. (a) Treatment-irradiated wound at 40X magnification. (b) Control-left wound at 40X magnification. (c) Treatment- irradiated wound at 100X magnification. (d) Control-left wound at 100X magnification. White arrows indicate a boundary between the normal dermis and wound gap.	28
Figure 3.9 H&E staining of wound biopsies on 10 days after surgery. (a) Treatment-irradiated wound at 40X magnification. (b) Control-left wound at 40X magnification. (c) Treatment- irradiated wound at 100X magnification. (d) Control-left wound at 100X magnification. White arrows indicate a boundary between the normal dermis and wound gap.	29
Figure 3.10 H&E staining of wound biopsies on 22 days after surgery. (a) Treatment-irradiated wound at 40X magnification. (b) Control-left wound at 40X magnification. (c) Treatment-irradiated wound at 100X magnification. (d) Control-left wound at 100X magnification. White arrows indicate a boundary between the normal dermis and granulation tissue. 	30
Figure 3.11 The maximum value of breaking load on 2 months after surgery: (a) four conditions analyzed by by using Kruskal-Wallis H test, (b) two groups compared by using Mann-Whitney U test. Asterisk represents the statistical difference (p<0.05) between two groups.	32
Figure 3.12 The maximum value of breaking load on 2 months after surgery: (a) four conditions analyzed by by using Kruskal-Wallis H test, (b) two groups compared by using Mann-Whitney U test. Asterisk represents the statistical difference (p<0.05) between two groups.	33 

                                    

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