Gene expression of bone marrow cells from rats on HAP blocks and β-TCP blocks

HAP 阻滞剂和 β-TCP 阻滞剂上大鼠骨髓细胞的基因表达

• 批准号: 17592013
• 负责人: HORIUCHI Rumi
• 金额: $ 2.18万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17592013/
• 关键词: HAP; B-TCP; scaffold; cell culture; micro array; 移植・再生医療; 細胞・組織; 歯学

In this study, investigation of osteogenesis around HAP and β-TCP blocks, cell culture on the scaffold using with HAP and B-TCP blocks were carried out and gene expression profiles of them were examined for the purpose of the reconstruction of jaw bone using with HAP and β-TCP blocks.HAP and β-TCP blocks were used for scaffolds and standard plastic tissue culture were prepared for control. Bone marrow cells from rats cultured following Maniatopoulos, et. al. were seeded onto the materials and the control. 1, 3 and 5days after seeding, total RNA was isolated from HAP and B-TCP blocks or plastic surfaces. After c-DNA was synthesized from total RNA, labeled c-RNA was prepared, fragmented and added to a hybridization mixture using Rat Oligo Microarray Kit (Agilent Technologies). Washing and staining was done and images were acquired using the Agilent Technologies Microarray scanner. Gene Spring software (Agilent Technologies) was used to analyze the results.The expression of a variety of genes in HAP and β-TCP surfaces, as compared with the control group showed respective change characteristically. 2-fold up regulated genes of them were included in apoptosis group, bone formation or resorption-related group, cell cycle regulated group, transcription group and etc. Up regulated genes of B-TCP in early stage had the genes related to hydroxyapatite binding or calcium ion binding.From these results, it was suggested that investigation of osteogenesis around HAP and β-TCP blocks would be possible.

本研究对HAP和β-TCP块周围的成骨情况进行了研究，对HAP和B-TCP块在支架上的细胞培养进行了研究，并检测了它们的基因表达谱，目的是利用HAP和β-TCP块重建颌骨。采用HAP和β-TCP块作为支架，制备标准塑料组织培养物作为对照。将Maniatopoulos等人培养的大鼠骨髓细胞植入材料和对照组。播种后1,3和5d，从HAP和B-TCP块或塑料表面分离总RNA。从总RNA合成c-DNA后，制备标记的c-RNA，使用大鼠Oligo微阵列试剂盒（Agilent Technologies）将其片段化并加入杂交混合物中。清洗和染色完成，使用Agilent Technologies Microarray扫描仪获取图像。使用Gene Spring软件（Agilent Technologies）对结果进行分析。与对照组相比，HAP和β-TCP表面多种基因的表达均表现出各自的变化特征。其中2倍上调基因分为凋亡组、骨形成或骨吸收相关组、细胞周期调控组、转录组等。B-TCP早期上调的基因与羟基磷灰石结合或钙离子结合相关。从这些结果来看，研究HAP和β-TCP块周围的成骨是可能的。