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

本研究的目的是探讨结缔组织生长因子（CTGF）诱导的自体骨髓间充质干细胞（MSCs）移植用于牙槽骨再生的可行性.人骨髓细胞的分离和细胞鉴定骨髓细胞从志愿者的人髂骨骨髓中分离。将人骨髓细胞铺板并在含有10%FBS的DMEM中培养。第9代细胞经免疫组化染色鉴定为表达STRO-1的细胞，这些细胞具有多向分化（成骨细胞和成脂细胞）. CTGF对干细胞行为的影响。1）通过涂覆CCN 2/CTGF剂量依赖性地增强细胞在羟基磷灰石盘上的附着效率，CCN 2/CTGF可通过αvβ3整合素激活p38 MAPK信号通路; 2）CCN 2/CTGF可剂量依赖性地促进细胞增殖; 3）趋化细胞迁移实验表明CCN 2/CTGF可显著促进细胞迁移; 4）CCN 2/CTGF可通过α v β 3整合素激活p38 MAPK信号通路。CTGF不影响细胞向成骨细胞的分化; 5）CCN 2/CTGF还促进了内皮细胞的粘附和增殖; 6）体内植入研究表明，CCN 2/CTGF促进了干细胞的存活并诱导其迁移到多孔羟基磷灰石支架中。

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.