口腔医学研究

口腔医学研究 ›› 2016, Vol. 32 ›› Issue (6): 622-626.DOI: 10.13701/j.cnki.kqyxyj.2016.06.017

• 临床研究论著 • 上一篇    下一篇

牙本质细胞外基质蛋白对乳牙牙髓干细胞体外增殖、分化能力的影响

张宇娜1 ,张琦1 ,倪世磊2 ,赵菁2 ,高雪彬1 ,黄洋1* ,孙宏晨2   

  1. 1. 吉林大学口腔医院儿童口腔科 吉林 长春 130021;
    2. 吉林大学口腔医院病理科
  • 收稿日期:2015-10-16 出版日期:2016-06-26 发布日期:2016-06-22
  • 通讯作者: 黄洋,电话:0431-88796038
  • 作者简介:张宇娜(1989~ ),女,吉林人,医师, 硕士,主要从事牙本质细胞外基质、乳牙牙髓干细胞,釉质矿化及再生方向基础研究以及儿童口腔医学临床研究。
  • 基金资助:
    吉林省科技厅自然基金项目(编号:20160101040JC)

Effect of DEMPs on Proliferation and Differentiation of SHED in Vitro.

ZHANG Yu-na1, ZHANG Qi1, NI Shi-lei2, ZHAO Jing2, GAO Xue-bin1, HUANG Yang1*, SUN Hong-chen2.   

  1. 1. Department of Pediatric Dentistry, School and Hospital of Stomatology, Jilin University, Changchun 130021, China;
    2. Department of Oral Pathology, School and Hospital of Stomatology, Jilin University, Changchun 130021, China
  • Received:2015-10-16 Online:2016-06-26 Published:2016-06-22

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摘要： 目的:提取牙本质细胞外基质蛋白(Dentinextracellularmatrixproteins,DEMPs)研究其对人脱落的乳牙牙髓干细胞(Stemcellfromhumanexfoliateddeciduousteeth,SHED)体外增殖分化能力的影响。方法:采用组织块联合酶消化法获得SHED并进行体外培养。成骨诱导液诱导细胞,鉴定其多向分化能力;拔取健康牛牙,用4mol/L盐酸胍和0.5mol/LEDTA提取DEMPs,四唑盐比色法(MTT)检测不同浓度DEMPs对SHED增殖能力的影响,同时测定DEMPs对SHED碱性磷酸酶(ALP)活性的影响;实时荧光定量PCR(RT-PCR)检测牙本质磷蛋白与牙本质基质蛋白1的mRNA表达情况。结果:DEMPs诱导细胞培养3d、5d可促进细胞增殖。细胞培养5、7d上调ALP活性,RT-PCR结果显示,DEMPs诱导后可促进DSPP与DMP-1基因的表达。结论:牙本质细胞外基质蛋白可以促进SHED的增殖与牙向分化能力。

关键词: 牙本质细胞外基质蛋白, 乳牙牙牙髓干细胞, 增殖, 分化

Abstract: Objective: To investigate the effect of DEMPs on proliferation and differentiation of SHED in vitro and to provide theoretical basis for the application of DEMPs in tooth reparation and regeneration. Methods: The stem cells were collected from human exfoliated deciduous teeth by enzyme digestion and tissue ex-plant method. We assessed the multi-differentiation potentials of the cells by inducement. Healthy teeth wereextracted from cattles, and prepared dentine matrix of the teeth was extracted by 4mol/L guanidine hydrochloride and 0.5mol/L EDTA. The effect of DEMPs on the proliferation activity of SHED was evaluated by MTT method and the effect ofDEMPs on the activity of ALP was also examined. The expression of DMP-1 and DSPPinduced by DEMPs observed by RT-PCR. Results: The proliferation of SHED was induced by DEMPs after three and five days, and increased activity of ALP was induced by DEMPs after five and seven days. The mRNA expression of DSPP and DMP-1 in SHED induced by DEMPs. Conclusion: DEMPs can promote the proliferation and the odontogenic differentiation of SHED.

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