臺灣博碩士論文加值系統

在台灣，從2002年開始，糖尿病為十大病因的第4位。在住院的糖尿病患者中，有百分之二十五是因為糖尿病足住院，而造成糖尿病傷口癒合不良的原因是多重因素所造成的，諸如生長因子的缺乏、細胞功能的受損、微小血管的病變、以及血糖控制的失調等。糖尿病的微小血管病變，主要原因為血管異常生長和微小血管網路的受損，進而導致傷口不易癒合。
在傷口癒合的過程當中，血液供應是非常重要的，在最近的研究發現，在正常的狀況下，血管內皮的先驅細胞(progenitor cell)在發生缺血性傷害時，會從骨髓經由血液循環，到達傷口處參與血管新生及組織修復的進行，但是在糖尿病病患的身上，骨髓幹細胞從骨髓進行到受傷組織的能力有問題，進一步造成組織的增生與細胞分化的功能不良。為了改善這個問題，我們使用局部增加幹細胞的模式，來達到改善傷口癒合的目的。
幹細胞是一群間質分化前的細胞，可以自我保持更新、增長及具有分化成多種不同細胞的能力，包括：脂肪、軟骨、骨細胞…等等，也可以分泌多種不同的激素.，在人體的許多組織中可以發現幹細胞。我們選用脂源性幹細胞來進行實驗，因為在臨床上容易取得，手術併發症機會低，病患接受度高，可以培養出的幹細胞數量多，而且增生的代數多。
我們運用自體脂源性幹細胞皮下注射於Streptozotocin(STZ)誘發的糖尿病鼠的傷口周圍來進行傷口癒合研究模型。我們於傷口周圍皮下注射1x106/ml以Quantum Dot(QD) 655螢光物質標定的老鼠自體脂源性幹細胞，以注射phosphate buffered saline和沒有注射的傷口來做對照組實驗，我們進行傷口外觀的觀察並且把傷口取下進行切片及染色，包括H&E stain 及螢光染色，以光學顯微鏡及共軛焦顯微鏡進行觀測。在STZ誘發糖尿病鼠組的傷口外觀，第5、7天的傷口，脂源性幹細胞組的傷口癒合百分比，傷口的成熟度及微血管的數量皆較另外兩組高。在正常鼠組中，在傷口癒合百分比在第七天，脂源性幹細胞組較另外兩組高，有統計上的意義，在其他項目，則沒有顯著的統計意義。脂源性幹細胞聚集於傷口周圍的毛囊與微血管在螢光顯微下可以見到在注射幹細胞的傷口中，糖尿病鼠傷口中移行的幹細胞較正常鼠少，我們也觀察到脂源性幹細胞分化為血管細胞。脂源性幹細胞聚集於傷口周圍正常皮膚的毛囊與微血管附近。
在此研究中，脂源性幹細胞經由增加血管新生，細胞分化及移行促進老鼠傷口癒合，我們需要更多研究來了解幹細胞在傷口癒合所扮演的角色。

Diabetes mellitus is the 4th leading cause of death in Taiwan. The prevalence rate of DM in those who are over 40 year-old persons was 11 to 13 percent. Diabetic foot ulcer is one of the most common complications among DM patients. It account for 25% hospitalization in Kaohsiung Medical University Hospital. Impaired wound healing in DM patients is related to DM neuropathy, macroangiopathy, microangiopathy, and dysfunctioned macrophages.
Recently, research in the field of vasculogenesis had revealed that bone marrow- derived endothelial progenitor cells are mobilized in response to injury, migrate to distant healing wounds. Although the number of progenitor cells contained within the bone marrow niche in diabetic animals was found to be normal, their mobilization into the circulation and homing to peripheral sites of injury were severely impaired. Thus, we implant the adipose- derived stem cells (ADSCs) locally to promote wound healing.
Mesenchymal stem cells (MSCs), are referred as stromal progenitor cells, are self-renewing and expandable stem cells. They can differentiate into adipocyte, chondrocyte, and osteoblast. They can also secrete many cytokines. Thus, MSCs play an important role in regenerative medicine, including the wound healing model. The reasons that we choice ADSCs are easily accessibility, low donor site morbidity, high yielding rate of the stem cells, and good quality.
We performed autologous ADSCs implantation intradermally to enhance wound healing in streptozotocin (STZ)- induced DM rats with a full-thickness excisional wound model. We injected 1x106/cc ADSCs which were labeled with Quantum Dot(QD) 655 intradermly around the wounds. We injected phosphate-buffered saline or took non- injection wounds as comparative controls. We recorded the wound condition and calculated wound closure percentage. Wounds were harvested at day five and day seven and then processed, sectioned, stained (H&E and CD31) and observed by microscope and confocal microscope.
ADSCs- treated wounds exhibited significantly accelerated wound closure percentage, with increased re-epithelialization, wound maturation, and angiogenesis. There are no statistically significant findings in a normal rat wound model except for an increased wound healing percentage in ADSCs-treated group on day seven. We also observed that ADSCs differentiated into endothelial cells. The ADSCs aggregated around hair follicles and capillary in adjacent normal skin.
In this study, ADSCs accelerated wound healing via angiogenesis, ADSCs differentiation and migration in a STZ-induced DM rat wound model. We need more studies to understand the role of stem cells in wound healing process.

一、 中文摘要……………………………………………01
二、 緒論…………………………………………………03
三、 研究方法與材料……………………………………07
四、 結果…………………………………………………13
第一部份……………………………………………13
第二部份……………………………………………15
第三部份……………………………………………19
五、 討論…………………………………………………28
六、 展望…………………………………………………32
七、 英文摘要……………………………………………38
八、 參考文獻……………………………………………40
九、 附錄 :英文縮寫……………………………………46

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