Title Page
Contents
ABBREVIATIONS 8
국문 초록 10
ABSTRACT 13
CHAPTER Ⅰ. Identification of exosomal biomarkers, and its optimal isolation and detection method for the diagnosis of Parkinson's disease: A systematic review and meta-analysis 15
국문 초록 16
ABSTRACT 19
Ⅰ. INTRODUCTION 23
Ⅱ. MATERIALS AND METHODS 27
A. Search strategy 27
B. Study selection/selection criteria 27
C. Inclusion and exclusion criteria 28
D. Data extraction 29
E. Risk of bias/quality assessment 30
F. Statistical analysis 31
Ⅲ. RESULTS 32
A. Study selection 32
B. Risk of bias and quality assessment using the Joanna Briggs Institute (JBI) critical appraisal checklist for case-control and cohort studies 34
C. Characteristics of the eligible studies 34
D. Meta-analysis 46
E. Subgroup analysis, publication bias, meta regression and sensitivity analysis 54
F. Exosomal miRNAs in Parkinson's Disease 62
Ⅳ. DISCUSSION 64
Ⅴ. LIMITATIONS 69
Ⅵ. CONCLUSION 70
REFERENCES 71
APPENDIX 86
CHAPTER Ⅱ. Efficacy of Fasudil, Synuclean-D and Direct Blue-1 loaded exosome for the better treatment of Parkinson's disease 89
국문 초록 90
ABSTRACT 92
Ⅰ. INTRODUCTION 95
Ⅱ. MATERIALS AND METHODS 98
A. Cell culture 98
B. Isolation of exosomes from SH-SY5Y 98
C. Characterization analysis of exosomes 99
D. Preparation of Fasudil, Synuclean-D and Direct Blue-1 loaded into exosome 100
E. Characterization of drug encapsulated exosomes with NTA and TEM analysis 101
F. Cell viability assay 101
G. Experimental animals 102
H. Experimental groups and doses of drugs 102
I. Open-field test 103
J. Statistical analysis 103
Ⅲ. RESULTS 105
A. Exosome characterization 105
B. Characterization of drug loaded exosomes 109
C. Determination of cell viability 115
D. Open-field test 119
Ⅳ. DISCUSSION 123
Ⅴ. LIMITATIONS 125
Ⅵ. CONCLUSION 126
REFERENCES 127
CURRICULUM VITAE 130
CHAPTER Ⅰ 22
Table 1. Quality assessment of the included studies using the JBI critical appraisal tool for case-control studies. 36
Table 2. Quality assessment of the included studies using the JBI critical appraisal tool for cohort studies. 38
Table 3. Characteristics of the eligible studies were included in the systematic review and meta-analysis. 40
Table 4. Leave-one-out sensitivity analyses on exosomal α-synuclein to investigate the source of heterogeneity. 56
CHAPTER Ⅰ 21
Figure 1. Flowchart (PRISMA 2020) of study selection. 33
Figure 2. Forest Plot showing the Ratio of Mean (RoM) of exosomal α-synuclein between patients with Parkinson's disease and healthy controls. 48
Figure 3. Forest Plot showing the Ratio of Mean (RoM) of exosomal (a) Tau, (b) PS-129, (c) DJ-1/PARK-7, (d) clusterin between patients with Parkinson's... 49
Figure 4. Forest Plot showing the Ratio of Mean (RoM) of (a) optimal exosome types, and (b) optimal exosome sources for α-synuclein between patients with... 52
Figure 5. Forest Plot showing the Ratio of Mean (RoM) of (a) optimal exosome isolation method, and (b) optimal measuring method for α-synuclein between... 53
Figure 6. Funnel plot of the ratio of mean for exosomal α-synuclein in patients with Parkinson's disease showing publication bias with pseudo 95% CI. 57
Figure 7. Forest Plot showing the Ratio of Mean (RoM) of exosomal α-synuclein for the age group 60-69 and 70-79 between patients with Parkinson's disease... 58
Figure 8. Bubble plot with a fitted meta-regression line of the correlation between Ratio of Mean (RoM) of exosomal α-synuclein and the age of patients... 59
Figure 9. Forest Plot showing the Ratio of Mean (RoM) of exosomal α-synuclein for different locations of patients with Parkinson's disease and healthy controls. 60
Figure 10. Forest Plot showing the Ratio of Mean (RoM) of exosomal α-synuclein for the disease stage of patients with Parkinson's disease. 61
Figure 11. The flow of the study conducted to identify the potential exosome-derived diagnosis biomarkers, and the most suitable methods for isolation and... 70
CHAPTER Ⅱ 94
Figure 1. Characterization of isolated exosomes. 108
Figure 2. Characterization of different drug loaded exosomes. 112
Figure 3. The number of nano-particles before drug loading and after drug loading into exosomes. 113
Figure 4. TEM images of loaded (A) Fasudil (B) Syn-D (C) Direct Blue-1. 114
Figure 5. Determination of cell viability (%) with different treatments on (A) MPTP induced and (B) 6-OHDA induced SH-SY5Y cell line. 116
Figure 6. Physical properties of the cells after different drug treatments in the (A) MPTP and (B) 6-OHDA induced SH-SY5Y cell lines. 118
Figure 7. Determination of motor function by analyzing distance travelled after different treatments in MPTP induced mice. 120
Figure 8. Changes of motor function following treatments in C57BL/6J mice. 121
Figure 9. Determination of anhedonia by analyzing time spent in center after different treatments in MPTP induced mice. 122