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不同浓度的牙髓干细胞成骨能力的研究

钱石兵 张凌鹏 殷凌云 李昌全 李虎 于鸿滨

钱石兵, 张凌鹏, 殷凌云, 李昌全, 李虎, 于鸿滨. 不同浓度的牙髓干细胞成骨能力的研究[J]. 昆明医科大学学报, 2023, 44(2): 61-68. doi: 10.12259/j.issn.2095-610X.S20230214
引用本文: 钱石兵, 张凌鹏, 殷凌云, 李昌全, 李虎, 于鸿滨. 不同浓度的牙髓干细胞成骨能力的研究[J]. 昆明医科大学学报, 2023, 44(2): 61-68. doi: 10.12259/j.issn.2095-610X.S20230214
Shibing QIAN, Lingpeng ZHANG, Lingyun YIN, Changquan LI, Hu LI, Hongbin YU. Study on Osteogenic Ability of Dental Pulp Stem Cells at Different Concentrations[J]. Journal of Kunming Medical University, 2023, 44(2): 61-68. doi: 10.12259/j.issn.2095-610X.S20230214
Citation: Shibing QIAN, Lingpeng ZHANG, Lingyun YIN, Changquan LI, Hu LI, Hongbin YU. Study on Osteogenic Ability of Dental Pulp Stem Cells at Different Concentrations[J]. Journal of Kunming Medical University, 2023, 44(2): 61-68. doi: 10.12259/j.issn.2095-610X.S20230214

不同浓度的牙髓干细胞成骨能力的研究

doi: 10.12259/j.issn.2095-610X.S20230214
基金项目: 云南省生物医药重大科技专项基金资助项目(202102AA100007);昆明市卫生科技人才培养“百”工程学科带头人培养项目(2021-SW(省)-001);昆明市卫生健康委员会卫生科研课题项目(2021-08-02-001)
详细信息
    作者简介:

    钱石兵(1996~),男,云南昆明人,在读硕士研究生,主要从事牙周和口腔黏膜疾病的防治工作

    通讯作者:

    于鸿滨,E-mail: yuhongbin6310@163.com

  • 中图分类号: R285.5

Study on Osteogenic Ability of Dental Pulp Stem Cells at Different Concentrations

  • 摘要:   目的   探究不同浓度的牙髓干细胞(dental pulp stem cell,DPSC)在定量Bio-Oss骨粉中最佳的成骨比例。   方法   收集牙髓组织、组织块贴壁法分离培养原代细胞,流式细胞术鉴定DPSC表面标志物,诱导成骨及成脂鉴定DPSC多向分化潜能;GFP慢病毒转染DPSC,嘌呤霉素筛选GFP-DPSC;设置1×105、2×105、4×105、8×1054组细胞数接种于0.05 g骨粉并成骨培养,对照组不加骨粉,第3天和第7天进行碱性磷酸酶活性测定;培养第7天在扫描电镜下观察细胞在骨粉内的形态变化。   结果   原代分离培养成功的DPSC符合间充质干细胞来源,相关表面标记物高表达,可向成脂和成骨分化;GFP成功转染DPSC,第3天和第7天进行碱性磷酸酶活性检测,实验组均高于对照组,差异具有统计学意义(P < 0.05);扫描电镜发现DPSC在骨粉表面爬行,部分细胞伸出伪足,4×10 5和8×105 2组细胞较为密集。   结论   4×105~8×105个DPSC与0.05 g Bio-Oss骨粉复合培养成骨效果较好,可以缩短成骨周期,细胞若接种过多,成骨效果反而抑制,造成干细胞的浪费。
  • 图  1  DPSC的培养及多向分化能力鉴定

    A:DPSC原代细胞(×50);B:扩大培养第三代细胞(×50);C:DPSC成骨对照组组染色所见(×100);D:DPSC成骨诱导组染色所见(×100);E:DPSC成脂对照组染色所见(×200);F:DPSC成脂诱导组染色所见(×200)。

    Figure  1.  Culture of DPSC and appraisal of multi -directional differentiation ability

    图  2  流式鉴定DPSC干细胞标记物

    Figure  2.  Identification of DPSC surface antigen by flow cytometry

    图  3  GFP转染DPSC不同时间荧光表达(×100)

    A:36 h GFP转染DPSC的荧光表达;B:72 h GFP转染DPSC的荧光表达;C:96 h GFP转染DPSC的荧光表达。

    Figure  3.  Fluorescence expression of GFP transfected DPSC at different time

    图  4  GFP-DPSC在Bio-Oss骨粉中的生长(×100)

    A:第3天时GFP-DPSC在Bio-Oss骨粉中的状态;B:第14天时GFP-DPSC在Bio-Oss骨粉中的状态。

    Figure  4.  Growth of GFP-DPSC in Bio-Oss bone powder

    图  5  第3天ALP活性比较

    *P < 0.05,**P < 0.01。

    Figure  5.  Comparison of ALP activity on day 3

    图  6  第14天ALP活性比较

    *P < 0.05,**P < 0.01。

    Figure  6.  Comparison of ALP activity on day 14

    图  7  1×105组同一视野下SEM图像

    A:100 μm ;B:50 μm。

    Figure  7.  SEM images of 1×105 groups in the same field of view

    图  8  2×105组同一视野下SEM图像

    A:100 μm ;B:50 μm。

    Figure  8.  SEM images of 2×105 groups in the same field of view

    图  9  4×105组同一视野下SEM图像

    A:100 μm ;B:50 μm。

    Figure  9.  SEM images of 4×105 groups in the same field of view

    图  10  8×105组同一视野下SEM图像

    A:100 μm ;B:50 μm。

    Figure  10.  SEM images of 8×105 groups in the same field of view

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  • 收稿日期:  2022-12-08
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