Title Page
Contents
ABSTRACT 9
Ⅰ. INTRODUCTION 11
Ⅱ. MATERIALS AND METHODS 14
1. Study Design 14
2. Decellularized Bone Scaffold Preparation 16
3. Preparation of Bone Marrow-Derived Mesenchymal Stem Cells 16
4. Thawing and Preparation of Bone Marrow-Derived Mesenchymal Stem Cells 17
5. Animal Experiment Procedures 17
6. Histologic Analysis of Osteogenesis in Decellularized Bone Scaffolds 18
1) H&E, Masson Trichrome, and PAS Staining 18
2) Osteocalcin Immunofluorescent (IF) Staining 19
7. Histologic Analysis of Vascularization in Decellularized Bone Scaffolds by CD31 Immunohistochemical Staining 20
Ⅲ. RESULTS 21
1. Histologic Assessment of Osteogenesis in Decellularized Bone Scaffold 21
2. Vascularization of Decellularized Bone Scaffold 25
3. Degree of New Bone Formation in Osteocalcin IF Staining 27
4. Statistical Analysis 30
Ⅳ. CONCLUSION 31
Ⅴ. DISCUSSION 33
REFERENCES 37
ABSTRACT IN KOREAN 40
Table 1. Experimental groups. 14
Table 2. Blood vessels counting by anti-CD31 Immunohistochemical staining (mean ± standard deviation) 26
Table 3. Degree of change to osteon structure in the decellularized bone scaffold (%, mean±standard deviation) 29
FIGURE 1. Study design and different conditionings of decellularized bone scaffold. 15
FIGURE 2. Hematoxylin and Eosin staining comparison of experimental groups. 22
FIGURE 3. Masson trichrome staining comparison of experimental groups. 23
FIGURE 4. New bone formation and bone forming cells relating to osteogenesis. 24
FIGURE 5. Anti-CD31 immunohistochemistry staining comparison of experimental groups. 25
FIGURE 6. Number of new blood vessels comparison between the groups. 26
FIGURE 7. Osteocalcin immunofluorescence staining comparison of experimental groups. 28
FIGURE 8. Degree of change to osteon structure comparison between the groups. 29