Establishment of an autologous cell transplantion method using mesenchymal stem cells for alveolus bone regeneration.

本研究旨在探讨利用结缔组织生长因子（connective tissue growth factor， CTGF）进行自体间充质干细胞移植牙槽骨再生的可能性1。人骨髓细胞的分离及细胞特性。从志愿者髂骨髓中分离骨髓细胞。人骨髓细胞在含10%胎牛血清的DMEM中培养。此外，通过免疫染色从培养的第9代细胞中鉴定出表达STRO-1的细胞，这些细胞表现出多谱系分化（成骨细胞和成脂细胞）。CTGF对干细胞行为的影响。1)细胞在羟基磷灰石盘上的附着效率通过包裹CCN2/CTGF呈剂量依赖性增强，CCN2/CTGF通过αvβ3整合素激活p38 MAPK信号通路。2) CCN2/CTGF显著增加细胞增殖的剂量依赖性。3) Chemotaxicell迁移实验显示，CCN2/CTGF显著增加了细胞迁移。4) CCN2/CTGF不影响细胞向成骨细胞的分化。5) CCN2/CTGF对内皮细胞的附着和增殖也有促进作用。6)体内植入研究表明，CCN2/CTGF可提高干细胞存活率，诱导其向多孔羟基磷灰石支架内迁移。

The purposes of this research are to investigate the possibility of autologous mesenchymal stem cell transplantion method for alveolus bone regeneration using connective tissue growth factor (CTGF).1. Isolation of human bone marrow cells and cell characterization.Bone marrow cells were isolated from human iliac bone marrow of volunteer. Human bone marrow cells were plated and cultured in DMEM containing 10% FBS. Moreover, STRO-1 expressing cells were identified by immunostaining from culture of passage 9 cells, and these cells showed the multilineage differentiation (osteoblastic and adipogenic).2. Effects of CTGF on stem cells behavior.1) Cell attachment efficiency onto hydroxyapatite disks is enhanced by coating CCN2/CTGF dose-dependently, and CCN2/CTGF activated p38 MAPK signal pathway via αvβ3 integrin.2) CCN2/CTGF significantly increased the cell proliferation dose dependently.3) The migration assay using Chemotaxicell indicated that CCN2/CTGF significantly increased the cell migration.4) CCN2/CTGF did not affect cell differentiation to the osteoblast.5) CCN2/CTGF also enhanced the endothelial cell attachment and proliferation.6) In vivo implantation study indicated that CCN2/CTGF enhance the stem cell survival and induce their migration into the porous hydroxyapatite scaffold.