| Background Tooth loss is a common disease in humans, have significant effects on the patients of mastication, pronunciation, appearance and psychology. At present, the repair method of missing teeth are many, but the most close to the real teeth, but less durable method. Although the dental implant and alveolar bone can be better combined, but the lack of the natural tooth structure of periodontal tissue. Therefore, tooth regeneration has become a hotspot of international research in stomatology. However, to regenerate a complete teeth, need to break many bottlenecks, solve the difficult problem of a lot of regenerative medicine in common. In recent years, some scholars put forward the concept of biological roots, namely the use of tissue engineering method, to natural or artificial synthesis with spatial structure of biological materials as the carrier, will have a dental cells "planted" to the carrier, and provide the cell proliferation and differentiation of micro environmental factors, through cell adhesion, proliferation and differentiation, in vitro or in vivo formation has a root in the periodontal ligament structure. Simple root construction does not involve the shape and size of the control of the crown, and in tooth tissue engineering research has a unique advantage, if can regenerate a hasbiological roots of periodontal ligament, and on the basis of post crown restoration, is expected to become a kind of ideal repair tooth loss of regenerative medicine treatment. The root tissue engineering with biological activity entirely possible to replace the implant denture repair is widely carried out. Root regeneration is a key scientific problems of tooth regeneration, root regeneration realization has important significance for clinical application, for the crown to construct or tooth construction provides a new research method. Seed cells, scaffold materials and microenvironment is the core part of construction bioroot. At present, scholars using tissue engineering technology, odontogenic mesenchymal stem cells and scaffold has biological roots of periodontal ligament, cementum and alveolar bone in vivo successfully constructed. However, the application of the seed cells for dental stem cell. Although odontogenic mesenchymal stem cells have good odontogenic differentiation ability, but the cells can only obtained by extracting the normal teeth can, and the quantity is limited, so in the clinical obtain autologous odontogenic mesenchymal stem cells subject to certain restrictions. For example, in edentulous patients in how to get to these odontogenic cells is a may hinder the problem for future clinical application. In dental stem cells derived by restriction, explore the use of human have been successfully isolated with transdifferentiation capacity of a variety of somatic stem cells for tooth regeneration become the inevitable choice. How to induce non dental stem cell odontogenic differentiation, to establish definite effect and stable induction system is key to the regeneration of cell source to solve the problem of teeth. Bone marrow-derived mesenchymal stem cells(BMMSC) is a strong proliferative capacity and multilineage differentiation potential of cells, has been widely used in bone tissue engineering. In addition, BMMSC can differentiate into other tissues and even other plasticity of endoderm cells, and BMMSC can be easily obtained, can be obtained from the patient's own, to avoid the problem of immune rejection. This research is based on the above results, the h TDM composite BMMSCs to explore its Odontogenetic differentiation ability. Provide the basis for tissue regeneration research project root.Aim:On the basis of the above research, this experiment using improved processing methods from tooth, the new tooth essence matrix scaffold materials can be, in vitro study of human dentin matrix materials on the effects of bone marrow mesenchymal stem cell biological characteristics and in vitro and in vivo research of human bone marrow mesenchymal stem cells in human dental essence matrix scaffold material, whether for differentiation of tooth. Study on root regeneration and lay the foundation for future clinical application.Materials and methods: 1 this experiment with bone marrow from clinical positive surgical repair jaw facial deformity patients, after the patient consent to collect bone tissue during operation and the remaining patients in mandible at. By using the method of tissue culture, bone marrow mesenchymal stem cells. 2. In this research, through the improved gradient de mining method was used for the treatment of human dentin matrix is obtained after the TDM, scanning electron microscope(SEM) dentinal tubules fully exposed, and tube and tube periodontal essential fibrous tissue became very loose. 3 by the method of flow cytometry for identification of the cell surface molecules. 4 using immunohistochemistry for identification of the source of this experiment cell. 5 by adipogenic and osteogenic experiment, this research obtained by the experiment of bone marrow mesenchymal stem cell differentiation 6 light microscope at TDM after bone marrow mesenchymal stem cells biological characteristics change. 7 using real time PCR detection by TDM extraction after induced by bone marrow mesenchymal stem tooth gene expression to cell. 8.h TDM composite BMMSCs transplanted into nude mice, in vivo experiments to explore the ability of TDM to explore BMMSCs' s teethResults:1 by using the method of tissue culture of jaw bone will cut inoculation broken bottles after 96 h was exchanged for the first time, after a week around the optical microscopic2 jaw tissues cells grow, mostly fusiform, polygon. Then the cells gradually increased. After passage, cells become more diversified. 2 we receive the fresh isolated tooth root production and the similar shape of TDM, about 1mm thick material, the surface morphology is smooth and regular. Ultrastructurally, the dentinal tubules were fully exposed, and between the tube and the peritubular dentin fibrous tissue became very loose. 3 cell culture production of BMMSCs was fixed by Vimentin and CK14 immunohistochemical staining to determine the cell source. Vimentin staining, CK14 staining positive negative 4 flow cytometry machine, detection of cell surface CD molecules. The indexes included: CD105, D31, CD34, CD90, CD73, CD25, CD26, CD146, CD8 b etc.. 5 human bone marrow mesenchymal stem cells into adipocytes and osteoblasts experiments showed that human bone marrow mesenchymal stem cells have strong osteogenic and adipogenic ability. 6 h TDM of human bone marrow mesenchymal stem cells, TDM facilitates cell growth, cell number, cell adhesion on the surface of the material. 7 extracts induced by h BMSCs 7 days after PCR quantitative detection of TDM, odontogenic related gene expression significantly increased. 8 h TDM composite BMMSCs transplanted subcutaneously into nude mice after 2 months, found that TDM surface structure of mineralization.Conclusion: 1 using tissue culture method to isolate and culture bone marrow mesenchymal stem cells. 2 bone marrow mesenchymal stem cells derived from the expression of related proteins in epithelial markers, expression of mesenchymal cell origin, and the expression of stem cell characteristics. 3.TDM can promote bone marrow mesenchymal stem cells Odontogeneticdifferentiation. |
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