Title Page
Contents
ABSTRACT 11
Ⅰ. Total Introduction 13
Ⅱ. In Vitro Determination of the Skin Anti-Aging Potential of 3D Human Adipose-Derived Mesenchymal Stem Cell Conditioned Medium 16
2.1. Introduction 16
2.2. Materials and Methods 19
2.2.1. Cell lines and cell culture 19
2.2.2. 3D ADMSC-CM cream preparation 22
2.2.3. Cell growth 24
2.2.4. Colony forming unit (CFU) measurement 24
2.2.5. Cytotoxicity test 25
2.2.6. Immune response analysis 25
2.2.7. Analysis of growth factors 26
2.2.8. RT-qPCR for EGF and VEGF 26
2.2.9. TYR activity inhibition test 27
2.2.10. Melanin inhibition test 27
2.2.11. Confirmation of HA expression 27
2.2.12. Assessment of expression of wrinkle-related factors 28
2.2.13. Statistical analysis 29
2.3. Results 30
2.3.1. Cell growth of HDFs 30
2.3.2. Cell proliferation of 3D ADMSC-CM 34
2.3.3. Immune response of 3D ADMSC-CM 36
2.3.4. Secretion of growth factors in 3D ADMSC-CM 38
2.3.5. mRNA expression of EGF and VEGF 40
2.3.6. Confirming the inhibitory effect of TYR 42
2.3.7. Melanin contents 44
2.3.8. HA expression effect 46
2.3.9. Evaluation of the antioxidant effect 48
2.3.10. Measurement of collagen expression level 51
2.4. Conclusion 53
Ⅲ. Clinical Utility of Cream containing 3D Human Adipose-Derived Mesenchymal Stem Cell Conditioned Medium 55
3.1. Introduction 55
3.2. Materials and Methods 62
3.2.1. Test participants 62
3.2.2. Skin measurement 63
3.3. Results 70
3.3.1. Reporting skin adverse reactions by participant survey 70
3.3.2. Evaluating alleviation of skin irritation by external stimuli (chemical stimuli) and the soothing effect 72
3.3.3. Evaluating the soothing effect on the irritated skin by physical stimuli (friction) 75
3.3.4. Evaluating refinement of rough skin texture damaged by external stimuli 78
3.3.5. Evaluating improved exfoliation by external stimuli (damage) and the soothing effect 81
3.3.6. Assessment of the efficacy on skin lifting 84
3.3.7. Assessment of skin elasticity and resilience 88
3.3.8. Skin tone and radiance evaluation 92
3.3.9. Skin freckle evaluation 95
3.3.10. Evaluation of the moisturizing effect of 3D ADMSC-CM under extreme cold and dry weather conditions 97
3.4. Conclusion 100
Ⅳ. Discussion 102
References 109
Appendix 126
Abstract (in Korean) 131
〈Table 1-1〉 Composition of cream-type cosmetics containing 3D ADMSC-CM 23
〈Table 2-1〉 Big data analysis on keywords for stem cell condition media cosmetics 57
〈Table 3-1〉 Skin adverse reactions reported by the testes (N=22) 71
〈Table 4-1〉 Results of skin lifting measurement 85
〈Table 5-1〉 Mean of improvement rate (%) of each person by variables of skin elasticity and skin resilience 89
〈Table 6-1〉 The rate of subjects with improved skin elasticity and skin resilience by variables 91
〈Figure 1-1〉 Acquisition of 3D stem cell conditioned medium. 21
〈Figure 2-1〉 Effects of 3D adipose-derived mesenchymal stem cell conditioned medium on cell growth. 33
〈Figure 3-1〉 Effects of 3D ADMSC-CM on cell proliferation. 35
〈Figure 4-1〉 Effects of 3D adipose-derived mesenchymal stem cell conditioned medium on IL-1α in LPS-stimulated human epidermal keratinocyte. 37
〈Figure 5-1〉 Secretion of growth factors affecting skin elasticity in stem cell conditioned medium. 39
〈Figure 6-1〉 mRNA expression of EGF and VEGF in by 3D ADMSC-CM. 41
〈Figure 7-1〉 Inhibitory effect of 3D ADMSC-CM on tyrosinase activity. 43
〈Figure 8-1〉 Effects of 3D ADMSC-CM on the melanin contents. 45
〈Figure 9-1〉 Effect of hyaluronan (HA) production by HDF after treatment with 3D ADMSC-CM. 47
〈Figure 10-1〉 Effects of antioxidation of 3D ADMSC-CM cream. 50
〈Figure 11-1〉 Effects of 3D ADMSC-CM cream on the secretion of Collagen 1. 52
〈Figure 12-1〉 Comparison of soothing and improvements in skin irritation by an external stimulus (chemical stimulation). 73
〈Figure 13-1〉 Soothing effect on physical skin irritation (friction). 76
〈Figure 14-1〉 Analysis of rough skin damage induced by external stimuli. 79
〈Figure 15-1〉 Improved exfoliation and soothing of skin damaged by external stimuli. 82
〈Figure 16-1〉 Image analysis of the lifting effect of the 3D ADMSC-CM cream. 86
〈Figure 17-1〉 Analysis of skin tone and skin gloss by 3D ADMSC-CM cream. 93
〈Figure 18-1〉 Improvement in skin freckles after the application of the cream containing three-‐dimensional adipocyte-derived mesenchymal... 96
〈Figure 19-1〉 Effect of moisturizing by 3D ADMSC-CM cream. 98