Title Page
Contents
Abstract 12
Chapter 1. Facile Fabrication of Microfiber-templated Microfluidic Chips with Microfibrous Channels for High Throughput and Continuous Production of Nanoscale Droplets and liposome 15
Abstract 16
Introduction 18
Materials and methods 22
Results and discussion 27
Conclusion 56
Reference 58
Chapter 2. Fabrication of Multiple-splitting Microfluidic Device with Microfibrous Channels for Continuous Production of Lipid Nanoparticles 63
Abstract 64
Introduction 66
Materials and methods 70
Results and discussion 78
Conclusion 103
Reference 105
Chapter 3. Fabrication of Microfluidic Bioreactor with Fibrous Micromixers for Continuous mRNA Synthesis 110
Abstract 111
Introduction 112
Materials and methods 117
Results and discussion 127
Conclusion 153
Reference 155
Abstract 164
Table 1-1. MTMC fabrication conditions. 50
Table 2-1. Fabrication variables for the microfibrous matrices with different diameters. 102
Table 3-1. Preparation variables for the microfibrous matrices with different diameters. The distance between the syringe nozzle (16 G) and... 150
Table 3-2. Composition of the solution used for one IVT reaction. 151
Table 3-3. mRNAs percent identity table synthesized in bulk and PFPE-MBR. 152
Fig. 1-1. MTMC fabrication procedure. 39
Fig. 1-2. SEM images of microfiber and MTMC. 40
Fig. 1-3. Size of MTMC junction. 41
Fig. 1-4. Time-lapse fluorescent microscopy images. 43
Fig. 1-5. Confocal images. 44
Fig. 1-6. Image and size of the water droplets in the outlet channel of MTMC. 45
Fig. 1-7. CV values. 46
Fig. 1-8. Color change of Nile red and UV absorbances. 48
Fig. 1-9. Size and encapsulation efficiency of the PEG-DA. 49
Fig. 2-1. MDMC fabrication procedure. 91
Fig. 2-2. SEM analysis of 3D printed channel and microfiber. 92
Fig. 2-3. Characterization of LNP fabricated using MDMC. 93
Fig. 2-4. Stability of LNPs. 94
Fig. 2-5. Schematic illustration and Fluorescence microscopy images of MDMCs. 95
Fig. 2-6. Cellular uptake efficiency of pyrene-loaded LNPs. 97
Fig. 2-7. Characterization of DOX-LNPs. 98
Fig. 2-8. Cytotoxicity of DOX-LNPs by concentration. 99
Fig. 2-9. Cytotoxicity and Live/Dead staining of DOX-LNPs. 101
Fig. 3-1. Fabrication procedure and SEM images of PDMS-based micromixers 139
Fig. 3-2. Characterization of micromixers. 140
Fig. 3-3. Mixing efficiency of MBR. 142
Fig. 3-4. Mixing behavior in micromixer. 143
Fig. 3-5. Continuous in vitro transcription in PFPE-MBR. 145
Fig. 3-6. Mixing profile of PFPE-MBR. 147
Fig. 3-7. Integrity test and performance evaluation of mRNA. 148
Fig. 3-8. mRNA sequencing. 149
Video 1-1. Microfibrous channel during DCM evaporation. 51
Video 1-2. Introduction of the water into the thin MTMC12. 52
Video 1-3. Generation of the water droplets in the thin MTMC12. 53
Video 1-4. Produced water droplets flowing in the outlet channel of MTMC12. 54
Video 1-5. Produced water droplets flowing in the outlet channel of the various MTMCs. 55