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研究生: 廖子瓏
Liao, Tzu-Lung
論文名稱: 開發可攜帶多重抗生素與即時偵測菌落群生長的新型智慧水凝膠敷料
Development of a new smart hydrogel dressing that can load multiple antibiotics and detect colony growth in real time
指導教授: 吳炳慶
Wu, Ping-Ching
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: 2021
畢業學年度: 109
語文別: 英文
論文頁數: 61
中文關鍵詞: 燒燙傷抗生素抑菌測試超音波衰減係數超音波成像菌落群生物膜
外文關鍵詞: Burn wounds, Wound healing, Antibiotics, Antibacterial test, Ultrasonic attenuation coefficient, Ultrasonic imaging, Bacterial colony, Biofilm
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  • 世界衛生組織估計,每年發生了1100萬例各種類型的燒燙傷,全世界每年有180,000人死亡。燒傷後的感染仍然是所有年齡層燒傷患者中最常見的並發症,並且是死亡最常見原因,佔燒傷相關死亡率60%至75%。
    燒傷面積和患者的年齡決定了燒傷患者死亡率的重要決定因素。隨著年齡的增加,較大的燒傷傷口和傷口的真菌和細菌的存在顯著提高了死亡率。最常見的死因是多重系統器官衰竭,而傷口感染引起的死亡率為45.9%。科學研究發現,燒燙傷傷口存在許多細菌並形成菌落群,進而生成生物膜。生物膜通常附著在組織或傷口表面,它們對抗生素和宿主天然免疫細胞具有高度的抗性,必須透過標準微生物培養來批量檢測,無法立即檢測。
    隨著科學技術的進步,超過3000種產品在針對燒燙傷傷口的治療過程中的各個方面都已經得到了發展。當前材料包括抗生素、人工皮、軟膏、水凝膠、海藻酸鹽敷料、膠原蛋白和玻尿酸。水凝膠已被證明具有許多有用的功能,並且最有利於燒傷患者的應用。因此,本文的目標是將三個不同的抗生素結合海藻酸鹽水凝膠製成一個智慧敷料,將Neomycin sulfate、 Bacitracin、 和polymyxin B sulfate結合於海藻酸鹽水凝膠,以預防傷口感染及吸收組織液效果,同時透過超音波及時偵測菌落群是否生長,達到早期預防生物膜生成效果。在材料特性上測試了藥物包覆率、傅立葉紅外線光譜儀分析、拉伸測試、形態分析、吸水率量測、抑菌測試、細胞相容性測定。智慧敷料結合超音波,測試材料衰減係數、超音波標準假體量測及透過超音波影像即時偵測菌落群生長。
    實驗結果顯示,新型智慧敷料具有較高的藥物包覆率(> 90%),形態分析的研究表明多孔結構的實現決定了傷口敷料吸收組織液的能力,同時具有生物相容性(> 80%的細胞生存力),在抗菌測試達到世界衛生組織所標示的臨界值徑規範,抑菌效果顯然能夠保護傷口免受微生物侵害。在超音波衰減係數量測中,與人工皮相比顯示具有較低的衰減係數,並且可以透過智慧敷料直接成像,達到及時偵測菌落群生長效果。

    The World Health Organization estimates that 11 million burns and scalds of various types occur every year, and 180,000 people die every year worldwide. Infection remains the most common complication and the most common cause of death among burn patients of all ages, accounting for 60 to 75 percent of burn-related mortality.
    The burn area and the patient's age are important determinants of the mortality for burn patients, in addition, there are many other factors. With the increase of age, larger burn wound, the presence of fungi and bacteria in wounds significantly increases the mortality rate. The most common cause of death seems to be multiple system organ failure, and 45.9% death caused by wound infection. The study found that there were many bacteria and colony formation in burn wound, which could produce biofilm. Biofilm is usually attached to the surface of tissues or wounds. They have high resistance to antibiotics and host natural immune cells. They must be detected in batch through standard microbial culture and cannot be detected immediately.
    With the advancement of science and technology, more than 3,000 products have been developed in all aspects of the treatment process for burn wounds. Current materials include antibiotics, artificial skins, ointments, hydrogels, alginate dressings, collagen, and hyaluronic acid. Hydrogels have been shown to have many useful functions and are most beneficial for burn patients. Therefore, the goal of this article is to combine three different antibiotics with alginate hydrogel to make a smart dressing, combining Neomycin sulfate, Bacitracin, and polymyxin B sulfate in the alginate hydrogel to prevent wound infection and absorption of tissue fluid effect. At the same time, ultrasound can detect the growth of bacterial colony in time, so as to achieve the effect of early prevention of biofilm formation.
    The material properties were tested for drug encapsulating efficiency, Fourier-transform infrared spectroscopy analysis, tensile test, morphological analysis, water absorption rate measurement, antibacterial activity, and cell compatibility. The smart dressing combined with ultrasound was used to measure the attenuation coefficient of the material, the standard prosthesis and the colony growth was detected by ultrasound image.
    The experimental results show that novel smart dressing has a high drug encapsulating rate (> 90%). The morphological analysis shows that the realization of the porous structure determines the ability of the wound dressing to absorb tissue fluid, and it has biocompatibility (> 80% of the cells viability). When the antibacterial test reaches the critical value of the World Health Organization, the antibacterial effect can obviously protect the wound from microbial invasion. In the ultrasonic attenuation coefficient measurement, it has a lower attenuation coefficient compared with artificial skin, and it can be directly imaged through the novel smart dressing to achieve the effect of detecting the growth of bacterial colonies in time.

    摘要 I Abstract III 致謝 V Contents VII List of Tables XI List of Figures XII List of Equations XIV Abbreviation List XV Chapter 1 Introduction 1 1.1 Burn wounds 1 1.1.1 Burn wound problems 1 1.1.2 Classification of burn wounds 1 1.1.3 Total body surface area injured 2 1.2 Mortality rate of burn wounds 2 1.3 Management of burn wounds 3 1.3.1 Inflammatory and infections of burn wounds 3 1.3.2 Bacterial growth in burn wounds 4 1.4 Prevent burn wound infection 4 1.5 Hydrogel is used in burn wound care 5 1.6 Ultrasound 6 1.6.1 Ultrasound wave 6 1.6.2 Attenuation coefficient 6 1.6.3 Ultrasound image 7 Chapter 2 Materials and Method 8 2.1 Materials 8 2.2 Experimental instruments 8 2.3 Fabrication of hydrogel loaded with multiple antibiotics 9 2.4 Determination of antibiotic content in hydrogels 9 2.5 The physicochemical properties of the hydrogel 10 2.5.1 Fourier transform infrared (FT-IR) spectrometry 10 2.5.2 Morphological analysis 11 2.6 Mechanical properties of hydrogels 11 2.6.1 Tensile testing of the hydrogel 11 2.7 Water absorption efficacy measurements 12 2.8 Antimicrobial analysis 12 2.9 Cell culture 13 2.9.1 NIH-3T3 cell culture 13 2.9.2 Cells counting 13 2.9.3 In vitro cell cytotoxicity assay 13 2.10 High frequency ultrasound imaging 14 2.10.1 Attenuation coefficients from each of the material 14 2.10.2 Wire phantom on material 15 2.10.3 HFUS B-mode image on human skin 15 2.10.4 Detection of bacteria colonies on HFUS B-mode image 16 Chapter 3 Results 17 3.1 Determination of antibiotic concentrations in Hydrogel 17 3.2 Characterization of the antibiotic hydrogel 17 3.2.1 Fourier‐transform infrared spectrum 17 3.2.2 Scanning electron microscope assay 18 3.3 Tensile testing of hydrogel 18 3.4 Hydrogel swelling ability test 19 3.5 Antibacterial activity 19 3.6 In vitro cytocompatibility of the antibiotic hydrogels 20 3.7 Attenuation coefficients of hydrogel and artificial skin 20 3.8 SNR and CNR on each material 20 3.9 An HFUS B-mode imaging of human skin 21 3.10 HFUS B-mode detects bacterial colonies with hydrogel 21 Chapter 4 Discussion 23 4.1 Fabrication of AH-F 23 4.2 AH-F to prevent burn wound infection 23 4.3 The Cell cytotoxicity assay 24 4.4 AH-F advantages compared with existing products 24 4.5 AH-F detects colony and prevents biofilm growth 25 4.6 Future application 26 Chapter 5 Conclusion 27 Tables and Figures 29 References 57

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