Bone Regeneration by Mesenchymal Stem Cell Sheets Overexpressing BMP-7 in Canine Bone Defects

학위논문 (박사)-- 서울대학교 대학원 : 수의학과, 2016. 8. 권오경.Successful repair of bone defect injuries is a major issue in reconstructive surgery. The necessary elements for bone healing include osteogenic cells, osteoinductive growth factors, and osteoconductive matrix. Multipotent mesenchymal stem cells (MSCs) and MSC sheets have potential for bone regeneration. Bone morphogenetic protein 7 (BMP-7) has been shown to possess strong osteoinductive properties. In addition, composite polymer/ceramic scaffolds such as poly ε-caprolactone (PCL)/β-tricalcium phosphate (β-TCP) are widely used to repair large bone defects. In the first chapter, the in vitro osteogenic potential of canine adipose-derived MSCs (Ad-MSCs) and Ad-MSC sheets are compared. Composite PCL/β-TCP scaffolds seeded with Ad-MSCs or wrapped with osteogenic Ad-MSC sheets (OCS) were also fabricated and their in vivo osteogenic potential was assessed after transplantation into critical-sized bone defects in dogs. The alkaline phosphatase (ALP) activity in osteogenic Ad-MSCs (O-MSCs) and OCS was significantly higher than that of undifferentiated Ad-MSCs (U-MSCs). ALP, runt related transcription factor 2 (RUNX2), osteopontin, and BMP-7 mRNA levels were up-regulated in O-MSCs and OCS compared to those in U-MSCs. The amount of newly formed bone was greater in PCL/β-TCP/OCS and PCL/β-TCP/O-MSCs/OCS than in the PCL/β-TCP/U-MSCs and PCL/β-TCP/O-MSCs groups. Although there was no difference between in vitro osteogenic genes expression of O-MSCs and OCS, the new bone formation was greater in the scaffold wrapped with OCS than the scaffold seeded U-MSCs and O-MSCs. Consequently, it was suggested that OCS could be used as an osteogenic matrix in canine critical-sized bone defects. OCS is difficult to handle and culture for more than two weekshowever, undifferentiated Ad-MSC sheets (UCS) are easy to culture and handle. The osteogenic capacity of USCs could be enhanced by canine BMP-7 gene transduction using a lentiviral system. Demineralized bone matrix (DBM), as defect filling and osteoinductive materials in large bone defects, was combined in a subsequent study. Combination of UCS overexpressing BMP-7 and DBM is supposed to be potential vehicles for bone healing. In the second chapter, canine Ad-MSCs overexpressing BMP-7 (BMP-7-MSCs) were produced and sheets formed from these cells (BMP-7-CS) were compared with Ad-MSC sheets for in vitro osteogenic potential. BMP-7-CS with and without DBM, were transplanted into critical-sized bone defects in vivo and osteogenesis was assessed. BMP-7 mRNA and protein levels were up-regulated in BMP-7-MSCs compared to those in Ad-MSCs. The ALP activity in Ad-MSC sheets and BMP-7-CS were significantly higher than that in Ad-MSCs. ALP, RUNX2, osteopontin, BMP-7, transforming growth factor-β and platelet-derived growth factor-β mRNA levels were up-regulated in BMP-7-CS compared to levels in Ad-MSCs and Ad-MSC cell sheets. BMP-7-CS showed the highest osteogenic abilities in vitro. The amount of newly formed bone and neovascularization were greater in BMP-7-CS and BMP-7-CS/DBM groups than in control groups. However, the BMP-7-CS/DBM group had more mineralized particles inside the defect sites than the BMP-7-CS group. As a result, it was suggested that a combination of BMP-7-CS and DBM could be used as osteogenic materials in canine critical-sized bone defects. BMP-7-CS not only provides BMP-7 producing MSCs but also produce osteogenic and vascular trophic factors. The combination of these cells with the osteoinduction and osteoconduction properties of DBM could result in synergy during bone regeneration. Thus, transplantation of BMP-7-CS and DBM could be used as an alternative treatment modality in bone tissue engineering.General introduction 1 CHAPTER I. Comparison of Osteogenesis between Adipose-Derived Mesenchymal Stem Cells and Their Sheets on Poly ε-Caprolactone/β-Tricalcium Phosphate Composite Scaffolds in Canine Bone Defects 4 Abstract 4 Introduction 6 Materials and methods 8 Results 19 Discussion 29 CHAPTER II. Osteogenic Potential of Adipose-Derived Mesenchymal Stem Cell Sheets Overexpressing Bone Morphogenetic Protein-7 in Canine Bone Defects 34 Abstract 34 Introduction 37 Materials and methods 40 Results 51 Discussion 64 Conclusion 68 References 70 Abstract in Korean 77Docto

The Status of Women in Islam

Since pre-Islanic times Arabian woman has been subject to her husband or nearest relatives. But the status of women greatly changed under Islam society although discrimination between man and woman made in pre-Islamic times still remains. In Islam society woman is allowed to inherit from her husband or relatives and control her property at her will. As far as other aspects of women"s life are concerned, the practices and traditions of pre-Islamic times still have an influence on the position of woman. Clearly the Koran adheres to the traditional view on woman as an inferior being(Koran 4,34). Islam permits man polygamy: e.g., a man can have as many as four wives. But in fact Koran promotes that one man should have a wife. Being considered a social contract, marriage is only able to be performed with man and womans mutual agreement. If a man wants to divorce, his wish is to be fulfilled by repeating his intention of divorce in public several times. But in case that woman wants to divorce, she can attain her purpose only after some conditions are satisfied. A divorced woman or a widow is allowed to remarry after some time passes. Womans wearing veil originated from the Koran is another testimony by which womans status can be estimated. Veil was a symbol of distinctions between man and woman. Closely connected with the exclusion of woman from public life is the veil. The traditionalists argue that woman should devote her entire life to keeping home by refering to the koran that man should take charge of his wife. They say that it is mans responsibility to earn his familys living because woman is unable to do outdoor works. The modernists claim that wearing of veil and womans living in seclusion are not genuine Islamic idea and that the purpose of the Koran is to establish its own reasonable ethical norms. The Koran says a great deal about the position of woman but most of them have been interpreted in such a way as means of socially restraining of woman or justifying mans right and his superior position. But womans status in Islam does not merely depend on the interpretation of the Koran. The traditional way of thoughts influence on their view on woman not less greatly than the Koran. In brief, the status of woman in Islamic society should be considered in terms of both the Koran and traditional views on woman

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