Gao Qianhua, Walmsley A Damien, Cooper Paul R, Scheven Ben A
School of Dentistry, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom.
School of Dentistry, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom.
J Endod. 2016 Mar;42(3):425-31. doi: 10.1016/j.joen.2015.12.019. Epub 2016 Jan 30.
Mesenchymal stem cells (MSCs) from dental tissues may respond to low-intensity pulsed ultrasound (LIPUS) treatment, potentially providing a therapeutic approach to promoting dental tissue regeneration. This work aimed to compare LIPUS effects on the proliferation and MAPK signaling in MSCs from rodent dental pulp stem cells (DPSCs) compared with MSCs from periodontal ligament stem cells (PDLSCs) and bone marrow stem cells (BMSCs).
Isolated MSCs were treated with 1-MHz LIPUS at an intensity of 250 or 750 mW/cm2 for 5 or 20 minutes. Cell proliferation was evaluated by 5-bromo-2-deoxyuridine (BrdU) staining after 24 hours of culture following a single LIPUS treatment. Specific ELISAs were used to determine the total and activated p38, ERK1/2, and JNK MAPK signaling proteins up to 4 hours after treatment. Selective MAPK inhibitors PD98059 (ERK1/2), SB203580 (p38), and SP600125 (JNK) were used to determine the role of activation of the particular MAPK pathways.
The proliferation of all MSC types was significantly increased after LIPUS treatment. LIPUS at a 750-mW/cm2 dose induced the greatest effects on DPSCs. BMSC proliferation was stimulated in equal measures by both intensities, whereas 250 mW/cm2 LIPUS exposure exerted maximum effects on PDLSCs. ERK1/2 was activated immediately in DPSCs after treatment. Concomitantly, DPSC proliferation was specifically modulated by ERK1/2 inhibition, whereas p38 and JNK inhibition exerted no effects. In BMSCs, JNK MAPK signaling was LIPUS activated, and the increase in proliferation was blocked by specific inhibition of the JNK pathway. In PDLSCs, JNK MAPK signaling was activated immediately after LIPUS, whereas p-p38 MAPK increased significantly in these cells 4 hours after exposure. Correspondingly, JNK and p38 inhibition modulated LIPUS-stimulated PDLSC proliferation.
LIPUS promoted MSC proliferation in an intensity and cell-specific dependent manner via activation of distinct MAPK pathways.
来自牙组织的间充质干细胞（MSCs）可能对低强度脉冲超声（LIPUS）治疗产生反应，这可能为促进牙组织再生提供一种治疗方法。本研究旨在比较LIPUS对啮齿动物牙髓干细胞（DPSCs）来源的MSCs、牙周膜干细胞（PDLSCs）来源的MSCs和骨髓干细胞（BMSCs）的增殖及丝裂原活化蛋白激酶（MAPK）信号传导的影响。
将分离出的MSCs用1MHz的LIPUS以250或750mW/cm²的强度处理5或20分钟。单次LIPUS处理后培养24小时，通过5-溴-2-脱氧尿苷（BrdU）染色评估细胞增殖。使用特异性酶联免疫吸附测定（ELISA）法测定处理后4小时内总的和活化的p38、ERK1/2及JNK MAPK信号蛋白。使用选择性MAPK抑制剂PD98059（ERK1/2）、SB203580（p38）和SP600125（JNK）来确定特定MAPK途径激活的作用。
LIPUS处理后，所有类型的MSCs增殖均显著增加。750mW/cm²剂量的LIPUS对DPSCs的影响最大。两种强度对BMSCs增殖的刺激程度相同，而250mW/cm²的LIPUS暴露对PDLSCs的影响最大。处理后，ERK1/2在DPSCs中立即被激活。同时，ERK1/2抑制特异性调节DPSC增殖，而p38和JNK抑制无影响。在BMSCs中，JNK MAPK信号被LIPUS激活，JNK途径的特异性抑制阻断了增殖的增加。在PDLSCs中，LIPUS处理后JNK MAPK信号立即被激活，而暴露4小时后这些细胞中的磷酸化p38 MAPK显著增加。相应地，JNK和p38抑制调节LIPUS刺激的PDLSC增殖。
LIPUS通过激活不同的MAPK途径，以强度和细胞特异性依赖的方式促进MSCs增殖。
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