中国组织工程研究

中国组织工程研究 ›› 2015, Vol. 19 ›› Issue (32): 5097-5102.doi: 10.3969/j.issn.2095-4344.2015.32.003

• 干细胞移植 stem cell transplantation • 上一篇    下一篇

牵张力下mTOR/MMPs信号通路可促进间充质干细胞-成骨细胞系的迁移

杨子桧1，吴宝磊1，贾 森1，杨新杰1，单 春1，刘晓昌1，2，王 磊1，雷德林1   

  1. 1解放军第四军医大学口腔医院军事国家重点实验室口腔颌面外科，陕西省西安市  710032；
    2解放军第四军医大学学员旅12连5班2011级口腔专业，陕西省西安市  710032
  • 出版日期:2015-08-06 发布日期:2015-08-06
  • 通讯作者: 雷德林，硕士，主任医师，解放军第四军医大学口腔医院军事国家重点实验室口腔颌面外科，陕西省西安市 710032 并列通讯作者：王磊，博士，主治医师，解放军第四军医大学口腔医院军事国家重点实验室口腔颌面外科，陕西省西安市 710032
  • 作者简介:杨子桧，男，1988年生，山西省大同市人，汉族，2013年佳木斯大学毕业，硕士，主要从事牵张成骨及干细胞方面的研究。
  • 基金资助:

    国家自然科学基金项目(81270015)

Mechanical stretch promotes mesenchymal stem cell-osteoblast lineage migration through activation of mammalian target of rapamycin/matrix metalloproteinases signaling pathway

Yang Zi-hui1, Wu Bao-lei1, Jia Sen1, Yang Xin-jie1, Shan Chun1, Liu Xiao-chang1, 2, Wang Lei1, Lei De-lin1   

  1. 1Department of Oral and Maxillofacial Surgery, Hospital of Stomatology, Fourth Military Medical University of PLA, Xi’an 710032, Shaanxi Province, China; 
    2Department of Stomatology, Fourth Military Medical University of PLA, Xi’an 710032, Shaanxi Province, China
  • Online:2015-08-06 Published:2015-08-06
  • Contact: Lei De-lin, Master, Chief physician, Department of Oral and Maxillofacial Surgery, Hospital of Stomatology, Fourth Military Medical University of PLA, Xi’an 710032, Shaanxi Province, China Corresponding author: Wang Lei, M.D., Attending physician, Department of Oral and Maxillofacial Surgery, Hospital of Stomatology, Fourth Military Medical University of PLA, Xi’an 710032, Shaanxi Province, China
  • About author:Yang Zi-hui, Master, Department of Oral and Maxillofacial Surgery, Hospital of Stomatology, Fourth Military Medical University of PLA, Xi’an 710032, Shaanxi Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81270015

PDF

431

被引次数

3

摘要：

背景：目前对于牵张成骨促进间充质干细胞-成骨细胞系迁移的机制尚未明了，亟需基于这一机制的新的治疗方案，以提高牵张成骨临床疗效。
目的：观察牵张应力下间充质干细胞-成骨细胞系mTOR信号通路相关基因的表达以及细胞迁移能力。
方法：将12只SD大鼠随机分为2组：牵张组大鼠(n=6)进行右侧下颌骨牵张成骨，非牵张组大鼠(n=6)行同侧下颌骨截断后安置牵张器但不牵张，术后15 d进行免疫组织化学染色分析新生骨痂中p-mTOR表达变化。另外，对培养的健康SD大鼠下颌骨间充质干细胞进行体外牵张试验，施加(6%，4 h)的静息牵张力，对照组不施加牵张力，实时定量PCR、划痕实验等方法检测间充质干细胞-成骨细胞系mTOR信号通路相关基因的表达及细胞迁移能力的变化。
结果与结论：牵张组大鼠新生骨痂中的p-mTOR表达高于非牵张组(P < 0.05)。与非牵张组(0%，4 h)相比，牵张组(6%，4 h)间充质干细胞-成骨细胞系的 mTOR、Raptor、p70S6K、MMP-2、MMP-9、MMP-13基因表达升高，间充质干细胞-成骨细胞系迁移距离更远(P < 0.05)。结果提示牵张应力可能通过激活间充质干细胞-成骨细胞系的mTOR/MMPs信号通路，促进其迁移。

 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 干细胞, 骨髓干细胞, 下颌骨, 间充质干细胞, 大鼠, 骨生成, 牵张, 信号转导, 国家自然科学基金

Abstract:

BACKGROUND: Distraction osteogenesis is one of the most important tissue engineering technologies. However, the exact signaling pathway controlling mesenchymal stem cell-osteoblast lineage (MSC-OB) migration during distraction osteogenesis has not yet been elucidated. More efforts should be paid to make a full understanding of the mechanism on MSC-OB lineage migration, which can improve the clinical efficacy of distraction osteogenesis.
OBJECTIVE: To evaluate the effects of mechanical stretch on the ability of MSC-OB mobility and expression of mammalian target of rapamycin (mTOR) signaling pathway as well as matrix metalloproteinases (MMPs) in MSC-OB, and to make clear the mechanism by which controls MSC-OB migration during distraction osteogenesis.
METHODS: Twelve Sprague-Dawley rats were randomized into two groups: experimental group (n=6), an in vivo 
rat mandibular distraction osteogenesis model was established on the right side of rats; non-stretch group (n=6), only the mandibular resection was done but with no distraction osteogenesis. Immunohistochemical staining was used to detect phosphorylated mTOR expression in new osteotylus at 15 days after operation. In addition, an in vitro cell stretch model was made in the mandibular mesenchymal stem cells from healthy Sprague-Dawley rats under resting tension force (6%, 4 hours); no distraction was done in control group. The ability of MSC-OB mobility, the expression of mTOR, Raptor, p70S6K and MMPs were evaluated using experiment methods including immunohistochemistry staining, real-time PCR and scratch assay.
RESULTS AND CONCLUSION: The expression of phosphorylated mTOR in MSC-OB was upregulated in the mandibular bone callus of the stretch group than the non-stretch group (P < 0.05). In the in vitro experiments, MSC-OB applied with mechanical stretch (6%, 4 hours) showed elevated gene expression levels of mTOR, Raptor, p70S6K, MMP-2, MMP-9 and MMP-13 compared with the control group (0%, 4 hours). Meanwhile, MSC-OB in the experiment group (6%, 4 hours) showed a greater ability of mobility, as demonstrated by a farther distance after 48 hours of observation (P < 0.05). The present study suggests that the enhancement of MSC-OB mobility correlates with increase of the gene expression of MMPs and mTOR signaling pathway. Mechanical stretch may promote MSC-OB migration through activation of mTOR/MMPs signaling pathway. 

 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Mandible, Mesenchymal Stem Cells, Osteogenesis, Stress, Mechanical

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