Transcriptome sequencing analysis reveals the effect of combinative treatment with low‑intensity pulsed ultrasound and magnesium ions on hFOB1.19 human osteoblast cells.
Transcriptome sequencing analysis reveals the effect of combinative treatment with low‑intensity pulsed ultrasound and magnesium ions on hFOB1.19 human osteoblast cells.
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DOI：
10.3892/mmr.2018.9006
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发表时间：
2018-07
影响因子：
3.4
通讯作者：
Zhao D
中科院分区：
文献类型：
--
作者：
Zu H;Yi X;Zhao D
可生物降解镁材料被认为是理想的骨合成植入物。然而，临床应用已经证明是复杂的。这主要与将植入物的降解程度降低到人体可接受的范围，同时促进成骨或骨诱导有关。在本研究中，镁离子和低强度脉冲超声(LIPUS)联合应用于hFOB 1.19人成骨细胞，作为解决这一问题的潜在策略。基因芯片分析表明，镁和/或LIPUS处理hFOB1.19成骨细胞后，共有7,314个差异表达基因(DEG)和826个共享DEG。基因本体论分析表明，在镁和LIPUS联合处理的细胞中，DEGS具有丰富的功能注释，包括“伤口愈合”、“转化生长因子β受体信号通路”、“转录、DNA模板”、“受体复合体”、“核”、“SMAD蛋白复合体”、“DNA结合”、“金属离子结合”和“GTP酶激活活性”。值得注意的是，转化生长因子-β、丝裂原活化蛋白激酶和肿瘤坏死因子信号通路在镁和利培酮联合用药组优先过度表达，随后通过逆转录定量聚合酶链式反应证实了这一点。联合用药组与对照组相比，骨形态发生蛋白6、Nogin、骨形态发生蛋白受体(BMPR)1A、BMPR2和Smad5/8等促进成骨细胞矿化的基因表达明显上调。与对照组相比，联合治疗组促进迁移的相关基因如c-jun氨基末端激酶、双皮质素、帕西林和jun原癌基因AP-1转录因子亚单位表达上调。茜素红S染色对成骨细胞的形态观察和创面愈合试验支持了DEG数据，表明镁和LIPUS联合治疗对成骨细胞的矿化和迁移具有协同作用。此外，与镁进入相关的金属转运体基因显著上调，包括ATPase Na+/K+转运亚单位α1、细胞周期蛋白和CBS结构域的二价金属阳离子转运体2、K+电压门控通道亚家族J家族成员14、瞬时受体电位阳离子通道M家族成员7和Trp亚家族V成员2。综上所述，本研究结果表明，镁和LIPUS联合刺激可能通过转化生长因子-β、丝裂原活化蛋白K和肿瘤坏死因子信号通路对人成骨细胞成骨起协同作用，同时也促进镁的内流。本研究证实了LIPUS和镁联合治疗作为一种新的治疗策略在提高可生物降解镁种植体的成骨诱导、生物相容性和生物安全性方面的潜力。
Biodegradable magnesium (Mg) materials are considered ideal as osteosynthesis implants. However, clinical application has proven complex. This is primarily associated with the issue of reducing the extent of implant degradation to a range acceptable for the human body, while simultaneously enhancing osteogenesis or osteoinduction. In the present study, a combination of Mg ions and low-intensity pulsed ultrasound (LIPUS) treatment was applied in hFOB 1.19 human osteoblast cells as a potential strategy to resolve this issue. A total of 7,314 differentially expressed genes (DEGs) and 826 shared DEGs in hFOB1.19 osteoblast cells were identified by microarray analysis following treatment with Mg and/or LIPUS. Gene Ontology analysis demonstrated that among cells treated with a combination of Mg and LIPUS, DEGs were significantly enriched in various functional annotations, including ‘wound healing’, ‘transforming growth factor beta receptor signaling pathway’, ‘transcription, DNA-templated’, ‘receptor complex’, ‘nucleus’, ‘SMAD protein complex’, ‘DNA binding’, ‘metal ion binding’ and ‘GTPase activator activity’. Notably, the transforming growth factor (TGF)-β, mitogen-activated protein kinase (MAPK) and tumor necrosis factor (TNF) signaling pathways were preferentially overrepresented in the Mg and LIPUS combination group, which was subsequently confirmed by reverse transcription-quantitative polymerase chain reaction. Furthermore, genes involved in osteoblast mineralization promotion, including bone morphogenetic protein 6, noggin, bone morphogenetic protein receptor (BMPR)1A, BMPR2 and SMAD 5/8, were significantly upregulated following combination treatment compared with the control group. Genes involved in the promotion of migration, including c-Jun N-terminal kinase, doublecortin, paxillin and Jun proto-oncogene AP-1 transcription factor subunit, were also upregulated in the combination treatment group compared with the control group. The DEG data were supported by morphological observations of the osteoblasts using alizarin red S staining and wound healing assays, which indicated that Mg and LIPUS combinative treatment had a synergistic effect on osteoblast mineralization and migration. Additionally, the combined treatment significantly upregulated metal transporter genes associated with Mg entry, including ATPase Na+/K+-transporting subunit α1, cyclin and CBS domain divalent metal cation transport mediator 2, K+ voltage-gated channel subfamily J member 14, transient receptor potential cation channel (TRP) subfamily M member 7 and TRP subfamily V member 2. In summary, the findings of the present study revealed that combined stimulation with Mg and LIPUS may exhibit a synergistic effect on human osteoblast bone formation through the TGF-β, MAPK and TNF signaling pathways, while also facilitating Mg influx. The present study demonstrated the potential of combinative LIPUS and Mg treatment as a novel therapeutic strategy for enhancing the osteoinduction, biocompatibility and biosafety of biodegradable Mg implants.
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DOI：
10.1080/09205063.2013.803455
发表时间：
2013
期刊：
Journal of biomaterials science. Polymer edition
影响因子：
--
作者：
Mountziaris PM;Dennis Lehman E;Mountziaris I;Sing DC;Kasper FK;Mikos AG
通讯作者：
Mikos AG
DOI：
10.1073/pnas.1018501108
发表时间：
2011-01-25
影响因子：
11.1
作者：
Glass, Graeme E.;Chan, James K.;Nanchahal, Jagdeep
通讯作者：
Nanchahal, Jagdeep
影响因子：
7.3
作者：
Osta B;Benedetti G;Miossec P
通讯作者：
Miossec P
影响因子：
4.8
作者：
McLaughlin, MM;Kumar, S;Young, PR
通讯作者：
Young, PR
影响因子：
4.4
作者：
Hannigan, MO;Zhan, LJ;Huang, CK
通讯作者：
Huang, CK