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利用微管陣列膜治療阿茲海默症作為新穎的封裝細胞治療法

為了解決Latency loaded vs un的問題，作者徐宗勤 這樣論述：

Table of contentsTable of contents 1Figures list 3Tables list 4致謝 5Acknowledgments 6摘要 7Abstract 8Graphical abstract 11Chapter II: Introduction 122.1 Neurodegenerative Disease 122.2 Alzheimer’s Disease 122.3 Familial Alzheimer’s Disease 132.4 Encapsulated cell therapy (ECT) 142.4.

1 Cell encapsulation categorizes into two scopes 142.4.2 Application of ECT in anti-cancer 172.5 Application of ECT in therapy for Alzheimer’s disease 182.6 Stem cell therapy for AD 192.7 Electrospinning 192.7.1 Microtube array membrane (MTAMs) 202.7.2 Application of MTAMs 202.7.3 MTA

Ms is a promising material for encapsulated cell therapy 21Chapter III: Materials and methods 263.1 Overview of materials and method 263.1.1 Preparation of MTAMs 263.1.2 Encapsulation cell therapy 263.1.3 Analysis 273.2 Preparation of PSF-PVP MTAMs 293.2.1 Characterization of MTAMs 30

3.2.2 SEM 303.3 In vitro study 303.3.1 Cell culture 303.3.2 Quantification of antibody 313.3.3 Cell loading within MTAMs 323.3.4 Cell viability(cytotoxicity) & proliferation 333.3.5 Live & Dead assay 353.4 In vivo study 353.4.1 Mice 363.4.2 Cell loading MTAMs implantation 363.4.3

Hybridoma cell characterization at the end of treatment 373.5 Determination of AD indicators in vivo 373.5.1 Immunohistochemistry 383.5.2 Western Blot 403.6 Animal behavior test 413.7 Statistical analysis 42Chapter IV: Results 434.1 Characterization of microtube array membrane (MTAMs

) by SEM 434.2 Hybridoma cell loaded MTAMs under a microscope 474.2.1 Hybridoma cell viability in a week was visualized by Live & Dead assay 484.2.2 Fluorescence mean intensity analyzed by ImageJ 504.2.3 Hybridoma cell viability in MTAMs by MTT 514.3 IgG2b released from hybridoma cell load

ed MTAMs, 75T Flask 524.3.1 IgG2b amount of empty MTAMs vs hybridoma cell loaded MTAMs (SC) in BALB/c mice serum 544.4 Morris water maze (MWM) 554.5 Passive avoidance 594.6 Immunohistochemistry of Hybridoma cell loaded MTAMs vs Empty MTAMs detected by Dako REAL EnVision/HRP secondary antibod

y 614.7 IgG2b detection in each group by Dako REAL EnVision/HRP secondary antibody (analyzed by TissueGnostics TissueFAXS SL) 624.7.1 IgG2b released from hybridoma cell loaded MTAMs to brain sections detected by Dako REAL EnVision/HRP secondary antibody (Analyzed by ImageJ) 674.8 P-Tau (Ser199

/Ser202) detection in each group (analyzed by TissueGnostics TissueFAXS SL) 704.8.1 The detection of AT8, P-Tau (Ser199/Ser202) on hippocampus & cortex (Analyzed by ImageJ) 764.9 AT8, P-Tau (Thr205) in hippocampus and cortex via western blot detection 774.10 3XTg AD mice weight after intracran

ial implantation of hybridoma cell loaded MTAMs & empty MTAMs 79Chapter V: Discussion 805.1 3D cell culture in MTAMs boosted the secretome 805.2 IgG2b released into blood circulations via ECT 805.3 The target site of phosphorylated tau for the efficacy of treatment 815.4 The relationship b

etween neuropathology and cognitive neuroscience in AD mice model 815.5 Hybridoma cell loaded MTAMs ameliorated AD in pathology and behavior change 825.6 The doubt of biosafety of hybridoma cell loaded MTAMs 84Chapter VI: Conclusion 85Chapter VII: Perspective 86Patent 87Reference 88Figure

s listFIGURE 1: THE GENERAL DEPICTION OF ENCAPSULATION CELL THERAPY FOR AD TREATMENT 11FIGURE 2: SCHEMATIC OF NANOGLAND AND INDEPENDENT SPACE FOR ISLETS IN DEVICE22 15FIGURE 3: THE CONCEPT OF MICROENCAPSULATED CELL THERAPY11 16FIGURE 4: MICROENCAPSULATION OF ISLETS 16FIGURE 5: HYBRIDOMA CELL SEC

RETED ANTI CEACAM6 MONOCLONAL ANTIBODY TO KILL MDA-MB-468 CANCER CELL LINE IN VIVO TEST.33 18FIGURE 6: THE WHOLE SCOPE OF FORMULATIONS FOR ENCAPSULATED CELL THERAPY 24FIGURE 7: ANALYSIS OF THE MORPHOLOGY OF MTAMS BY SCANNING ELECTRON MICROSCOPE. 30FIGURE 8: THE SCHEMATIC OF IGG2B QUANTIFICATION.

32FIGURE 9: A SCHEMATIC OF CELLS LOADED IN MTAMS. 33FIGURE 10: FLOW CHART OF CELL VIABILITY OF CELLS LOADED IN MTAMS BY MTT ASSAY. 34FIGURE 11: THE DIAGRAM OF THE EXPERIMENTAL DESIGN OF IN VIVO TEST. 36FIGURE 12: SCHEME OF HYBRIDOMA CELL LOADED MTAMS IMPLANTATION. 37FIGURE 13: CHARACTERIZATION

OF MICROTUBE ARRAY MEMBRANE (MTAMS) BY SEM. 44FIGURE 14: A FREQUENCY DISTRIBUTION OF MTAMS LENGTH AND WIDTH WITH THE FITTING OF A GAUSSIAN FUNCTIONAL CURVE. 44FIGURE 15: CHARACTERIZATION OF MTAMS LUMEN WALL. 45FIGURE 16: CHARACTERIZATION OF MTAMS POROUS DISTRIBUTION. 47FIGURE 17: HYBRIDOMA CELL

LOADED MTAMS WERE OBSERVED UNDER THE MICROSCOPE. 48FIGURE 18: LIVE & DEAD ASSAY OF HYBRIDOMA CELL LOADED MTAMS IN A WEEK. 49FIGURE 19: AVERAGE FLUORESCENCE INTENSITY (MEAN) = TOTAL FLUORESCENCE INTENSITY IN THE REGION (INTDEN) / AREA. 50FIGURE 20: MTT ASSAY ANALYZED HYBRIDOMA CELL VIABILITY IN MT

AMS. 51FIGURE 21: THE CUMULATIVE IGG2B CONCENTRATION OF EACH GROUP. 53FIGURE 22: THE IGG2B CONCENTRATION IN EACH GROUP AT THE ENDPOINT. 53FIGURE 23: IGG2B CONCENTRATION IN MOUSE SERUM. 54FIGURE 24: THE ESCAPE LATENCY AND TIME SPENT IN THE GOAL QUADRANT FOR EACH GROUP. 58FIGURE 25: THE TIME THAT

THE MICE ENTERED TO DARK COMPARTMENT BASELINE & AFTER 1.5 MONTHS. 61FIGURE 26: THE IMMUNOCHEMISTRY OF HYBRIDOMA CELL MTAMS. 62FIGURE 27: IGG2B DETECTION IN EACH GROUP. 65FIGURE 28: IGG2B POSITIVE SIGNAL IN A SINGLE CELL. 66FIGURE 29: BRIGHT-FIELD IMMUNOHISTOCHEMISTRY OF CORONAL MICE BRAIN (INTE

RAURAL: 5.7 MM, BREGMA: -3.3 MM). CHROMOGEN 3,3’-DIAMINOBENZIDINE (DAB) WAS OXIDIZED BY HORSERADISH PEROXIDASE (HRP) PRESENTING ON 2ND ANTIBODY AND BECAME BROWN COLOR PRECIPITATION. 68FIGURE 30: P-TAU (SER199/SER202) DETECTION IN EACH GROUP. 73FIGURE 31: PHOSPHO-TAU (SER199, SER202) IN A SINGLE CE

LL. 74FIGURE 32: BRIGHT-FIELD IMMUNOHISTOCHEMISTRY OF CORONAL MICE BRAIN (INTERAURAL: 5.7 MM, BREGMA: -3.3 MM). CHROMOGEN 3,3’-DIAMINOBENZIDINE (DAB) WAS OXIDIZED BY HORSERADISH PEROXIDASE (HRP) PRESENTING ON 2ND ANTIBODY AND BECAME BROWN COLOR PRECIPITATION. 76FIGURE 33: PHOSPHO-TAU (THR205) EXPR

ESSION IN THE WHOLE BRAIN WAS DIVIDED INTO CORTEX AND HIPPOCAMPUS. 78FIGURE 34: 3XTG AD MICE WEIGHT AFTER INTRACRANIAL IMPLANTATION OF HYBRIDOMA CELL LOADED MTAMS. 79Tables listTABLE 1:COMPARISON OF MTAMS WITH OTHER ENCAPSULATED CELL THERAPIES 23TABLE 2: THE SUMMARIZED REPORT OF IGG2B POSITIVE A

REA IN A SINGLE CELL FOR EACH GROUP. 65TABLE 3: THE SUMMARIZED REPORT OF P-TAU POSITIVE AREA IN A SINGLE CELL FOR EACH GROUP. 74

藉由點擊化學製備穀胱甘肽控制藥物釋放之胜肽微脂體

為了解決Latency loaded vs un的問題，作者鄭仁華 這樣論述：

中文摘要 iABSTRACT ii誌謝 iii目錄 iv圖目錄 vii表目錄 xi符號說明 xii一、 緒論（INTRODUCTION） 11-1 前言 11-2 微脂體 11-3 微脂體藥物的包覆與釋放 31-4 穀胱甘肽 61-5 以穀胱甘肽為引信響應的反應81-6 膜活性胜肽 121-7 胜肽微脂體的合成

方法 151-8 實驗動機與目的 16二、 實驗部分 (EXPERIMENTAL SECTION) 172-1 微脂體合成、定性與定量 17 2-1-1微脂體磷脂質濃度的定量 18 2-1-2藥物包覆率 19 2-1-3動態光散射分析 202-2 胜肽合成、定性與定量 20 2-2-1胜肽合成 20 2-2-2高效能液相層析法及質譜法 23 2-2-3胺基酸定量分析 24 2-2-4引信響應胜肽的還原反應

測試26 2-2-5圓二色光譜 262-3 胜肽微脂體合成、定性與定量 27 2-3-1後嵌入法 27 2-3-2胜肽與微脂體交聯反應-CuAAC28 2-3-3胜肽與微脂體交聯反應-SPAAC292-4 胜肽微脂體藥物釋放分析 30 2-4-1藥物引信釋放區間 30 2-4-2胜肽微脂體試管內藥物釋放 30三、 實驗結果與討論 (RESULT and DISCUSSION) 333-1 引信響應膜活性胜肽的設計及合成 333-2 胜肽的定性及定量

393-3 穀胱甘肽對引信響應胜肽的還原反應測試 413-4 胜肽微脂體製備方式的最佳化 45 3-4-1後嵌入法修飾藥物微脂體 46 3-4-2銅催化的疊氮-炔烴環加成反應 48 3-4-3張力促進的疊氮-炔烴環加成反應 503-5 胜肽微脂體引信釋放區間的評估 523-6 以穀胱甘肽為引信控制多肽微脂體試管內的藥物釋放 56 3-6-1不同比例的DSPE-PEG2000對引信釋放的行為影響 56 3-6

-2不同比例的錨定脂質對引信釋放的行為影響 59 3-6-3胜肽與微脂體交聯反應時間對引信釋放的行為影響 613-7 胜肽微脂體的定性 653-8 膜活性多肽之二級結構與多肽微脂體藥物釋放關係的探討 67四、 結論 (CONCLUSION) 70參考文獻 (REFERENCE) 71附錄 (Supporting Information)

80附錄一、 脂肽/疊氮化胜肽之合成轉換率分析 80附錄二、Imidazole-1-sulfonyl Azide Hydrochloride之合成 84附錄三、脂肽的還原反應測試實驗結果 84附錄四、膜活性脂肽對於DOPC微脂體的嵌入效率分析 89附錄五、胜肽/脂肽的HPLC圖譜與質譜圖 91