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研究生： Vinothini Arunagiri
Vinothini Arunagiri
論文名稱： Development of biopolymer composites absorbent in membrane and hemostatic application
Development of biopolymer composites absorbent in membrane and hemostatic application
指導教授： 蔡協致
Hsieh-Chih Tsai
口試委員: 賴君義
Juin-Yih Lai
鍾台生
Chung Tai-Shung
甄明阳
Jen-Ming Yang
李榮和
Rong-Ho Lee
學位類別： 博士
Doctor
系所名稱： 應用科技學院 - 應用科技研究所
Graduate Institute of Applied Science and Technology
論文出版年： 2021
畢業學年度： 109
語文別： 英文
論文頁數： 137
中文關鍵詞： 聚己內酯(PCL) 、聚-DL-乳酸(PDLLA) 、油吸附劑 、三聚氰胺 、分離 、明膠 、卡拉膠 、單寧酸 、微粒 、止血 、膨潤比
外文關鍵詞： PCL/PDLLA, Hydrophobic sorbent, K-Carrageenan, Microparticles,, Hemostasis,, swelling ratio
相關次數： 點閱：259  下載：1 
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    • 隨著世界的快速發展所衍生之汙染問題日趨嚴重，特別是水汙染方面更是對於自然生物影響巨大。 而多孔性合成薄膜由於生產成本低廉及所需體積小等優勢，在膜分離領域受到了廣泛的應用。其在設計較常見的分別為促進水運輸的親水性膜，以及抑制水運輸的疏水性膜，而後者又在氣體分離、氣體吸收、過濾、滲透蒸發及蒸餾等方面有更多元的應用。
    過去，研究人員對於水分離過程所應用之膜，更多關注在增加透水性以及汙垢控制方面，因此疏水性膜的開發也相對較少。而海中的原油洩漏，對於生物體的呼吸、進食及溫度調節等方面帶來了重大負面影響，是亟需解決的問題。因此，製備疏水且降解性多孔聚合物吸附劑，可進行對環境較無害之優勢，近期便受到了更多的重視及開發。
    在第一部分之研究中，使用冷凍乾燥造技術來對市售3D多孔三聚氰胺海綿的孔隙，進行生物可降解PCL/PDLLA (混摻比，90:10、85:15 & 80:20) 的填充，並評估原油洩漏處理的效果。在上述各比例的測試中，PCL/PDLLA (80:20) 改質三聚氰胺吸附劑在FE-SEM中觀察到了多孔互連的結構; 在接觸角分析中展現出較高的疏水性 (162° (WCA) & 180° (UOWCA)) ，並且具備超親油的濕潤性。此外，三聚氰胺海綿和PCL/PDLLA混合物之間的各項物理化學相互作用力也通過FT-IR、拉曼及XRD進行析，同時亦使用了拉伸試驗，並顯示出92%的伸長率。而PCL/PDLLA（80:20）改性三聚氰胺吸附劑對不同的黏度的油如正己烷、正辛烷、大豆油、原油和機油等，表現出約3.3-8.8 g/g的吸油能力。此外，PCL/PDLLA (80:20) 改性三聚氰胺吸附劑在重力作用下對油包水 (W/O) 乳液的分離也有99.5 %的去油效率，通量約為560.737 L m-2h-1。因此，可生物降解的PCL/PDLLA（80:20）改性三聚氰胺吸附劑可以作為一種較為環保的漏油處理吸附劑和油包水 (W/O) 乳液的去油劑，同時具備較佳的化學穩定性、機械穩定性和重複利用性。
    另一存在的嚴重性議題，為全球全因戰爭、事故或手術等狀況，所造成的不受控出血問題,其涵蓋約全球死亡率的40%。因此，具備生物相容性的止血材料的開發也在持續進行中，現階段雖已有許多研究,但能進行快速凝血的仍較少。而若將其製備為粉末型態，則能更好地適應不規則形的傷口，及大面積的醫療止血等優勢。
    在第二個研究中，是要通過製備粉末微粒來促進凝血之過程。是透過使用吐溫80(Tween 80) 的油包水(W/O) 乳化體系，來製備使用氫鍵及聚電解質作用力，所構成之生物相容性的明膠-單寧酸-κ-卡拉膠（GTC）微粒。製備的GTC微粒，其在46微米尺寸下，具備了約273%的體積膨脹率。 在NIH 3T3的細胞測試中表面出大於80%的生物相容性，溶血性試驗中表現出小於5%的血液相容性，同時伴隨較低的凝血指數。值得重視的是，GTC微粒能在50秒內誘導BALB/c雌性小鼠股動脈快速止血，失血量為46毫克，明顯優於對照組（凝血時間：250秒；出血量：259毫克），並在解決出血控制方面展現出了臨床應用之潛力。

    Global development of pollution in water becomes serious issue in the current world and has a great impact on living organisms, especially in ocean waters organisms. Due to the low production cost and small occupied space, the porous membrane has been widely used in the membrane separation process. Especially, porous polymeric membranes are developed with high hydrophilicity or hydrophobicity either to generate or prevent the passage of water, respectively. The vast applications oriented to hydrophobic membranes are filtration, gas separation, gas absorption, pervaporation, and distillation.
    Since, the researcher major focal point was to enhance the water permeability and fouling control in aqueous separation. The existence of hydrophobic membrane becomes relatively low in the past. In other hand, consequence of heavy oil spillage (ex. Crude oil) in ocean creates major pollution and escorts physical smothering like respiration, feeding, and thermoregulation to the living organism. Hence, recently, the existence of hydrophobic polymeric sorbent occurs and provided to be a significant resource to clean heavy oil spills. Especially, the eco-friendly polymer acts as a requisite compostable oil sorbent, due to the beneficiary degradation of non-toxic elements in water resource.
    Thus, I have designed my first experiment to investigate oil spill cleaning application using three-dimensional commercial porous melamine sponge’s substrate, which was modified with pore-filling eco-friendly PCL/PDLLA polymer blended agent in various ratios such as 90:10, 85:15 & 80:20, through dip coating process followed by inexpensive freeze-drying technique. In the above-fabricated sorbent ratios, PCL/PDLLA (80:20) sorbent displayed reliable emergence of interconnected porous structure in the FE-SEM morphology analyzer. Further, the surface wettability of PCL/PDLLA (80:20) modified sorbent have executed high super-oleophilic wettability along with greater hydrophobicity with a water contact angle in air at 162°(WCA) and under oil at 180°(UOWCA) in water angle analyzer. Furthermore, PCL/PDLLA modified sorbent’s physicochemical characteristics was determined through spectroscopic techniques like Fourier Transform-Infra Red and Raman mapping spectroscopy. Despite the previous characteristics, the crystalline and mechanical behavior of PCL/PDLLA (80:20) modified sorbent was confirmed through X-ray diffraction study along with 92% elongation in tensile test. Additionally, the fabricated PCL/PDLLA (80:20) modified sorbent exhibited moderate oil absorption capacity of about 3.3-8.8g/g in various viscous oils such as n-hexane, n-octane, soybean oil, crude oil, and motor oil. In addition, PCL/PDLLA (80:20) modified melamine sorbent also escorted 99% oil rejection efficiency with the flux of about 560 L m-2h-1 under gravity for W/O emulsion separation. Therefore, I would like to emphasize that biodegradable PCL/PDLLA (80:20) modified melamine sorbent can act as an eco-friendly oil spill cleaning sorbent and oil rejector from W/O emulsion with suitable chemical, mechanical stability, and reusability.
    Here comes another global serious issue is excessive uncontrolled bleeding during battlefield, civilian accidents, and surgery which causes humans to account for 40% mortality worldwide. Hence, the development of biocompatible hemostatic materials to control hemorrhage has become a fundamental research problem in the biomedicine community. Though existence of various form of hemostatic material occurs, still lacks in executing rapid coagulation cascade. Consequently, the use of powder materials in hemostasis has been emerged with several benefits like adaptability to different irregular wounds, large surface area, improved efficacy, and safety in medical uses.
    Hence, in our second study, we aimed to accelerate an improved coagulation cascade through powder microparticles. Therefore, we prepared biocompatible gelatin-tannic acid-κ-carrageenan (GTC) microparticles via polyelectrolyte interactions as well as hydrogen bonding from tannic acid using a facile Tween 80 stabilized water-in-oil (W/O) emulsion method. In addition, the formulated GTC microparticles were optimized and displayed a change in volume-swelling ratio of about 273% for a particle size of 46 µm. Subsequently, GTC microparticles have exhibited >80% biocompatibility in NIH 3T3 cells, <5% hemocompatibility in hemolysis ratio tests along with lower blood coagulation index. Notably, the GTC microparticles induced rapid hemostasis in 50 s with blood loss of ⸟46 mg in the femoral artery of BALB/c female mice, which was significantly better than the control group (blood clot time:250s; blood loss: 259 mg). Thus, our study findings propose that GTC microparticles may play a promising clinical role to tackle hemorrhage control

    摘要 ii ABSTRACT iv ACKNOWLEDGMENTS vii TABLE OF CONTENT viii LIST OF FIGURES xi LIST OF TABLES xiii LIST OF SCHEMES xiv LIST OF ACRONYMS xv CHAPTER 1 16 1.Introduction 16 1.1 Background of the study 17 1.1.1. First study 17 1.1.2. Second study: 19 CHAPTER 2 22 2. Literature survey 22 2.1: Impact of Global Issues on mortality of Living organisms 22 2.1.1: Water pollution 22 2.1.2: Hemorrhage: 23 2.2: Resolving water pollution and hemorrhage: 24 2.2.1. Membrane separation processes 24 2.2.2: Hemostasis and Steps of Hemostasis: 27 2.2.3: Hemostatic agent 29 2.3: significance of Porous and microparticles form of absorbent in water purification and hemorrhage control 33 2.3.1. Mechanism of porous membrane and microparticles in separation process and hemostasis: 36 2.4: Biopolymer as 38 CHAPTER 3 43 3. Objective of the Study 43 3.1. General objective 43 3.2. Specific objectives 43 3.3 Chemicals, Reagents and Instruments 44 3.3.1. Chemicals and Reagents 44 3.3.2. Instruments 45 3.4 Statistical Analysis 46 CHAPTER 4 47 4.Eco-friendly PCL/PDLLA absorbent in Oil-Spill Cleaning and Emulsified Oil separator 47 4.1. Introduction 47 4.2. Experimental Section 51 4.2.1. Fabrication of PCL/PDLLA melamine sorbent via dip-coating freeze dry method 52 4.2.2. Physicochemical characteristics 52 4.2.3. Stability of the PCL/PDLLA melamine sorbents 53 4.2.4. Oil absorption experiment in oil spill cleaning 53 4.2.5. Gravity driven surfactant stabilized W/O emulsion preparation and separation 54 4.2.6. Repeatability/Reusability 54 4.3. Results and Discussion 54 4.3.1. Optimized PCL/PDLLA Composition 54 4.3.2. Electron microscopy structural view of PCL/PDLLA Melamine sorbent: 57 4.3.3. Physicochemical characteristic of PCL/PDLLA Melamine sorbent: 59 4.3.4. PCL/PDLLA melamine act as sorbent in oil absorption experiment and chemical stability. 65 4.3.5. PCL/PDLLA Melamine act as membrane in gravity driven water in oil emulsion separation 69 4.3.6. Repeatability/reusability test in PCL/PDLLA (80:20) Melamine Sorbent 71 4.3.7. Wettability Performance in PCL/PDLLA (80:20) melamine sorbent 72 4.4. Conclusion 74 CHAPTER 5 80 5. Polyelectrolyte Gelatin-Tannic acid-κ-Carrageenan microparticle absorbent as hemostatic agents 80 5.1. Introduction 80 5.2. Experimental Section 82 5.2.1 Fabrication method of polysaccharide microparticles 82 5.2.2 Volume swelling properties of the microparticles 84 5.2.3 Rehydration rate 84 5.2.4 In vitro cytotoxicity assays 85 5.2.5 Hemocompatibility 86 5.2.6. Hemostatic Assay in vitro 87 5.2.7 Hemostasis in vivo 87 5.3. Results and Discussion 89 5.3.1 Optimization of Gelatin, Tannic acid, and Carrageenan components to obtain stable GTC microparticles 89 5.3.2 Morphology, poly electrolyte interaction, and surface charge examination in GTC microparticle: 92 5.3.3 Volume swelling ratio and rehydration rate of GTC microparticles: 96 5.3.4. Hemocompatibility of GTC microparticle: 100 5.3.5 Hemostasis in vitro 102 5.3.6 Biocompatibility and live cell/ dead cell imaging of GTC microparticles 103 5.3.7 In vivo femoral artery potential hemorrhage control of GTCm microparticle: 106 5.3.8: In vivo wound healing effects, healing rate and histopathology study: 109 5.4 Conclusions 115 CHAPTER 6 116 6.General Summary and Future Perspectives 116 6.1. Summary 116 6.2. Future perspectives and recommendations 117 REFERRENCES 118 Appendix 127 Appendix A: Supplementary Information: 127 Appendix B: Publications: 137 Appendix C: International Conference: 138

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