| Jaw defect is highly common, which is produced by trauma, infection, tumor and congenital disease. At present the main approach to the repair of the jaw defect is autoalleigic bone graft,and the bone graft is frequently obtained from ilium. In embryology, different from the trunk appendicular skeleton derived from mesoderm, most of the craniofacial bones are derived from the dorsal margins of the neural folds.The way of jaw bone development is intramembranous ossification and ilium is endochondral ossificatiom. Data from clinical test and animal experiment had indicated that in desected jaw defect the jaw-derived graft survival rate is higher and the absorption rate is lower than other bone-derived grafts.All the data show osteogenitor cells in different organs of the same individual have distinct biological characteristics.Currently the cell model of repairing bone defect is based on long bone and little is known about jaw bone marrow cells. The purpose of this study was to compare the biological characteristics of bone marrow stromal cells(BMSCs), obtained from rat mandible and ilium respectively. In vitro their biological characteristics were compared including cell proliferation,colony formation, osteogenic induction,minerlization ,and adipogenic induction. The results and concludions may provide new approach to the further investigation to the distinctive biological mechanism of jaw development and jaw defect repair. PART I Culture of BMSCs cells from rat mandible and iliumObjective: To isolate BMSCs of rat mandible and ilium by the approach relying on their ability to adhere to plastic surfaces.Method: Mandible and ilium from Sprague-Dawley maleness rat of 1 month, weighted 150±10g were sawn and gelatinous marrow were extracted to be suspended in DMEM and to be cultured. then their morphological characters and changes were observed.Result: Primary cells were plated in culture dishes and after 24h the cells adhering to plastic surfaces can be observed. Nonadherent cells are removed 24-72 h later by changing the medium. Following removal of nonadherent cells 1-4 days after the establishment of the culture, cells are maintained with periodic passages until a relatively homogeneous population of morphologically and immunophenotypically similar mesenchymal stem cells is established. Two types of morphology can be observed—I: elongated, fibroblastoid cells or II:large, flat cells. The adherent cells form colony-forming unit-fibroblast( CFU-F ) and cultures were first passaged after 2 weeks. Subculturing cells have high proliferation before passage-6 in ilium and passage-9 in mandible, then their growth rate decreases significantly, with part of subculturing cells demonstrating vacuole and particles in endochylema and resulting in apoptosis. Conclusion: BMSCs are isolated successfully which can be used for the latter experiments.PART II In vitro biological characteristics of BMSCs from ratmandible and iliumObjective: To explore the biological characteristics of BMSCs obtained from rat mandible and ilium respectively in vitro condition.Method: The biological characteristics of mandible and ilium-derived MSCs from passages 2 were compared, including growth curve, cell proliferation, osteogenic induction and adipogenic differention.Result: Compared to ilium-derived BMSCs, Mandible BMSCs proliferate more rapidly when plated at low density and can form more calcium accumulation than iliac cells. While ilium-derived BMSCs has higher capability of adipogenesis in the adipogenic supplements.Conclusion: BMSCs in different organs of the same individual have distinct biological characteristics. The feature of mandible BMSCs may provide some findings to the further research into pathological changes and local repair and regeneration of mandible. |
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