Title Page
Contents
ABSTRACT 9
1. Introduction 11
2. Materials and methods 14
2.1. Preparation of scaffolds 14
2.2. Fourier transform infrared spectroscopy (FTIR) 17
2.3. Scanning electron microscopy (SEM) 17
2.4. Water absorption test 17
2.5. Compressive stress test 18
2.6. Isolation of adipose derived stem cells 18
2.7. Cell culture method 19
2.8. Indirect cytotoxicity test 19
2.9. Lactate dehydrogenase (LDH) activity assay 20
2.10. Cell viability assay 20
2.11. Live/Dead-staining 21
2.12. Scaffold cell seeding process and media cell counting 21
2.13. Cell proliferation test 22
2.14. Field emission scanning electron microscope (FE-SEM) 22
2.15. Staining of scaffold 23
2.16. Statistical analysis 23
3. Results 24
3.1. Protein-polysaccharide interactions in the formation of protein-polysaccharide hydrogels and scaffolds 24
3.2. The effect of protein concentration on the properties of protein-polysaccharide hydrogel and scaffolds 26
3.3. Structural properties of protein-polysaccharide hydrogels 28
3.4. Physical characterization of plant-based hydrogel scaffolds 30
3.5. Impact of protein-polysaccharide scaffold leachates on cytotoxicity and cell viability 32
3.6. Characterization of cell attachment and proliferation on protein-polysaccharide scaffolds 35
3.7. Appearance of scaffold after staining 37
4. Discussion 39
5. Conclusion 47
References 48
Abstract (in Korean) 57
Table 1. The formulation of protein-polysaccharide hydrogels and scaffolds 16
Figure 1. Gel formation of the mixture of protein and polysaccharide solution and its scaffold. (A) Hydrogel formation of gels mixed with 2.5% proteins (soy and pea)... 25
Figure 2. Hydrogel and scaffold formation with different concentration of protein solutions. (A) Hydrogel formation of protein-polysaccharide solution mixed with... 27
Figure 3. Fourier-transform infrared (FT-IR) spectra of the protein-polysaccharide hydrogels. Protein-polysaccharide hydrogels were prepared by mixing with 2.5%... 29
Figure 4. Physical properties of protein-polysaccharide scaffolds. (A) Water absorption properties of the protein-polysaccharide scaffolds. (B) Compressive... 31
Figure 5. Cytotoxicity of protein-polysaccharide scaffolds. Adipose tissue-derived stem cells (ADSCs) from Berkshire piglet were treated with day 1 and day 3... 33
Figure 6. Cell adhesion properties of protein-polysaccharide scaffolds. (A) ADSCs on scaffolds captured by field emission scanning electron microscope (FE-SEM). 1... 36
Figure 7. Photographic comparison of unstained and stained scaffold. (A) Scaffold soaked in PBS for 15 min at room temperature. (B) Scaffold stained with beet... 38