Characterization of ROCK inhibitor-treated extracellular vesicles and analysis of their effect on osteogenesis

ROCK抑制剂处理的细胞外囊泡的表征及其对成骨作用的影响分析

• 批准号: 22K17135
• 负责人: Thant Lay
• 金额: $ 2.91万
• 依托单位: Niigata University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Early-Career Scientists
• 财政年份: 2022
• 资助国家: 日本
• 起止时间: 2022-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-22K17135/
• 关键词: Extracellular vesicles; ROCK inhibitor; Bone regeneration

MagCapture Exosome Isolation Kit was used to prepare Control extracellular vesicles (Control-EVs) and ROCK inhibitor-treated extracellular vesicles (ROCKin'-EVs) from the culture medium of BMSCs. Mass spectrometry was performed to elucidate the changes in the proteomic composition. We detected there were changes ROCKin'-EVs protein composition. We identified that a number of Rab GTPase (known as the key regulators of intercellular vesicle transport) were upregulated in ROCKin'-EVs.We isolated the Control-EVs and ROCKin'-EVs from the culture medium of BMSCs again using total exosome isolation aiming to collect large amount of EVs. Western blotting of Control-EVs and ROCKin'-EVs was performed. The quantity of isolated EVs was inconsistent and necessary to be normalized.

使用MagCapture外泌体分离试剂盒从骨髓间充质干细胞培养基中制备对照细胞外囊泡（Control- ev）和ROCK抑制剂处理的细胞外囊泡（ROCKin'- ev）。质谱分析分析了蛋白质组学组成的变化。我们检测到ROCKin -EVs蛋白组成发生了变化。我们发现，在ROCKin' s - ev中，许多Rab GTPase（被称为细胞间囊泡运输的关键调节因子）被上调。我们再次从骨髓间充质干细胞培养基中分离出control - ev和ROCKin'- ev，目的是收集大量的ev。对Control-EVs和ROCKin -EVs进行免疫印迹检测。分离出的ev数量不一致，需要进行归一化。

项目成果

Extracellular matrix profiling of cultured PDL cells by chemical digestion-assisted proteomics

通过化学消化辅助蛋白质组学对培养的 PDL 细胞进行细胞外基质分析

• 发表时间: 2022
• 作者: Lay Thant;Masaru Kaku;Azusa Dobashi;Yoshito Kakihara;Isao Saito;Katsumi Uoshima
• 通讯作者: Katsumi Uoshima

Alteration of the ECM composition and fiber organization in periodontal ligament under mechanical loading

机械负荷下牙周膜 ECM 成分和纤维组织的改变

• 发表时间: 2022
• 作者: Lay Thant;Masaru Kaku;Yoshito Kakihara;Masaru Mizukoshi;Megumi Kitami;Kohei Kitami;Daiki Kobayashi;Yutaka Yoshida;Takeyasu Maeda;Katsumi Uoshima;Isao Saito;Makio Saeki
• 通讯作者: Makio Saeki