A EXPERIMANTAL STUDY OF CHANGES IN SYNTHESIS OF MMPs AND TIMPs FOR BONE RESORPTION ON ORTHODONTIC TREATMENT

正畸治疗中骨吸收MMPs和TIMPs合成变化的实验研究

• 批准号: 09672129
• 负责人: GOTO Shigemi
• 金额: $ 2.3万
• 依托单位: AICHI-GAKUIN UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09672129/
• 关键词: ORTHODONTIC TREATMENT; TOOTH MOVEMENT; BONE RESORPTION; THREE-DIMENSIONAL CELL CULTURE; MMPs; TIMPs; MECHANICAL STRESS

Using newly developed ASAHI-KASEI cellulose porous sheets (ACPS) and Micro carrier (MC) as an artificial matrix, three-dimensional cell cultures were examined to see how these affected the expression of gelatinases and TIMPs(TIMP-1 and TIMP-2) in human osteosarcoma cell line MG-63 cells. Though this was only a preliminary experiment, a study was also made on how mechanical stress affects the viability of cells cultured three dimensionally in ACPS and MC. And, the expression level of TIMP-1 was analyzed by RT-PCR. The results are summarized as follows :1. MG-63 cells produced the same level of active gelatinase in either ACPS or a monolayer culture system. But, total gelatinase of the ACPS culture system was significantly higher compared with the monolayer culture system.2. The cells showed a tendency to produce higher levels of TIMPs in ACPS compared with those in monolayer.3. In either culture system, both active and proenzyme types of gelatinase were produced. But, only the cells in ACPS expressed gelatinase B together with gelatinase A.4. The cells showed a tendency to produce higher mRNA levels of MT1-MMP and TIMP-1, TIMP-2 in MC compared with those in monolayer by RT-PCR.5.The cells which added mechanical stress showed a tendency to produce higher mRNA levels of TIMP-1 in MC compared with those in control group by RT-PCR.From these results, it was concluded that the cells cultured in ACPS express their phenotypes differing qualitatively and quantitatively from those in monolayer culture, and that the three-dimensional culture system with ACPS can be used as an useful tool for in vitro orthodontic experiments.

使用新开发的旭化成纤维素多孔片（ACPS）和微载体（MC）作为人工基质，检查三维细胞培养物如何影响人骨肉瘤细胞系 MG-63 细胞中明胶酶和 TIMP（TIMP-1 和 TIMP-2）的表达。尽管这只是初步实验，但还研究了机械应力如何影响在 ACPS 和 MC 中三维培养的细胞的活力。并且，通过RT-PCR分析TIMP-1的表达水平。研究结果如下： 1． MG-63 细胞在 ACPS 或单层培养系统中产生相同水平的活性明胶酶。但ACPS培养体系的总明胶酶明显高于单层培养体系。 2．与单层细胞相比，ACPS中的细胞表现出产生更高水平TIMPs的趋势。3．在任一培养系统中，均产生活性明胶酶和酶原类型的明胶酶。但是，只有 ACPS 中的细胞同时表达明胶酶 B 和明胶酶 A.4。 RT-PCR结果表明，与单层细胞相比，MC中的细胞有产生较高MT1-MMP和TIMP-1、TIMP-2 mRNA水平的倾向。5。RT-PCR结果显示，与对照组相比，添加机械应力的细胞在MC中表现出产生较高TIMP-1 mRNA水平的倾向。从这些结果可以看出，在ACPS中培养的细胞表达的表型与在ACPS中培养的细胞在定性和定量上都有所不同。 单层培养，并且具有 ACPS 的三维培养系统可以用作体外正畸实验的有用工具。