Dynamic phase of Alveolar Bone-derived Osteoblasts at High Calcium Ion Concentration with Bone Resorption Accentuation.

高钙离子浓度下牙槽骨源性成骨细胞的动态阶段与骨吸收加速。

• 批准号: 11671887
• 负责人: MAENO Masao
• 金额: $ 1.92万
• 依托单位: Nihon-University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671887/
• 关键词: bone remodeling; high calcium ion concentration; matrix metalloprotease; alveolar bone; osteoclast differentiation factor; osteoclastogenesis inhibitory factor; osteoblast-like cells; tissue inhibitor of matrix metalloprotease; 破骨細胞分化因子; カルシウムイオン

Human alveolar bone-derived osteoblast-like cells were obtained from alveolar bone fragments of young (18-24 years old) and old (55-68 years old) patients. The cells that migrated from individual bone fragment were harvested separately and subcultured up to the third passage. After seeding the cells in 6-well microplates, the cells were cultured with α-MEM containing 10% FBS and 1.8 to 10 mM CaCl_2 for up to 72 h. The gene expression of extracellular matrix proteins, MMPs, TIMPs, osteoclast differentiation factor (ODF) and osteoclastogenesis inhibitory factor was examined at mRNA level by quantitative RT-PCR using specific primers. The following results were obtained.1. Although logarithmic cell proliferation was recognized in low Ca^<2+> concentration (1.8 and 2.5 mM), it was not recognized in high Ca^<2+> concentration (5 and 10 mM).2. Gene expression of bone sialoprotein and osteocalcin was increased by addition of high Ca^<2+> concentration in both age groups. Expression of type I collagen and osteonectin was not affected by addition of Ca^<2+> in both age groups.3. Expression of MMP-1 and MMP-2 was not affected by addition of Ca^<2+> in both age groups. MMP-3 expression was not detected with or without Ca^<2+> in both age groups.4. Expression of TIMP-1 and TIMP-2 was increased in Ca^<2+> concentration-dependent in both age groups, and the effects were recognized markedly in young age group.5. Expression of OCIF was increased by addition of high Ca^<2+> concentration in both age groups, and the effects were recognized markedly in young age group. In contrast, ODF expression was not detected with or without Ca^<2+> in both age groups.

人牙槽骨来源的成骨细胞样细胞从年轻（18-24岁）和老年（55-68岁）患者的牙槽骨碎片中获得。分别收获从单个骨碎片迁移的细胞，并传代培养至第三代。将细胞接种于6孔板中，用含10%FBS和1.8 ~ 10mMCaCl_2的α-MEM培养72 h。采用特异性引物定量RT-PCR检测细胞外基质蛋白、基质金属蛋白酶（MMPs）、基质金属蛋白酶抑制剂（TIMPs）、破骨细胞分化因子（ODF）和破骨细胞生成抑制因子（ODF）的mRNA表达。得到以下结果.尽管对数细胞增殖在低Ca^2+浓度（1.8和2.5 mM）下被识别，但在高Ca^2+浓度（5和10 mM）下不被识别。在两个年龄组中，骨唾液蛋白和骨钙素的基因表达都随着高浓度Ca^<2+>的加入而增加。在两个年龄组中，I型胶原和骨连接蛋白的表达不受Ca^<2+>的影响.在两个年龄组中，MMP-1和MMP-2的表达不受Ca^<2+>的影响。MMP-3在有或无Ca^<2+>的两个年龄组中均未检测到表达. TIMP-1和TIMP-2的表达在两个年龄组均呈Ca^<2+>浓度依赖性增加，且以青年组作用明显.高浓度Ca^<2+>对两个年龄组OCIF表达均有促进作用，且以青年组作用最为明显。相反，在有或无Ca^<2+>的两个年龄组中均未检测到ODF表达。