中国口腔种植学杂志 ›› 2021, Vol. 26 ›› Issue (4): 219-225.DOI: 10.12337/zgkqzzxzz.2021.08.003

• 论著·基础研究 • 上一篇    下一篇

帕米膦酸二钠对颌骨来源间充质干细胞成骨分化的影响

徐妍1,2, 孙晋3, 周海华4   

  1. 1深圳市罗湖医院集团口腔科,广东 518000;
    2武汉大学口腔医学院口腔基础医学省部共建国家重点实验室培育基地和口腔生物医学教育部重点实验室,湖北 430079;
    3南方医科大学附属深圳妇幼保健院口腔病防治中心,广东 518000;
    4武汉大学口腔医院口腔颌面外科,湖北 430079
  • 收稿日期:2021-06-25 出版日期:2021-08-10 发布日期:2021-09-08
  • 通讯作者: 徐妍,Email:635424216@qq.com,电话:0755-82200110
  • 作者简介:徐妍 口腔医学博士、主治医师,研究方向:口腔种植学,口腔颌面外科学
  • 基金资助:
    深圳市罗湖区软科学研究计划(LX20201205)

Effect of pamidronate disodium on osteogenic differentiation of jaw bone marrow mesenchymal stem cells

Xu Yan1,2, Sun Jin3, Zhou Haihua4   

  1. 1Department of Stomatology, Shenzhen Luohu Hospital Group, Guangdong 518000, China;
    2The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and the Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Hubei 430079, China;
    3Stomatology Health Care Center, Affiliated Shenzhen Maternity, Child Healthcare Hospital, Southern Medical University, Shenzhen University, Guangdong 518000, China;
    4Department of Oral and Maxillofacial Surgery, College and Hospital of Stomatology, Wuhan University, Hubei 430079, China
  • Received:2021-06-25 Online:2021-08-10 Published:2021-09-08
  • Contact: Xu Yan, Email: 635424216@qq.com, Tel: 0086-755-82200110
  • Supported by:
    Shenzhen Luohu District Soft Science Research Plan (LX20201205)

摘要: 目的 从颌骨来源间充质干细胞(jaw bone marrow mesenchymal stem cells, JBMMSCs)着手研究帕米膦酸二钠(pamidronate disodium,PD)对成骨分化的影响,从而探讨其引起骨坏死的原因。方法 采用噻唑蓝(MTT)试验在0、0.1、0.5、1.0、5.0、10.0 μg/ml浓度中筛选出适合JBMMSCs生长及后续研究的PD浓度;采用茜素红(ARS)染色定量、碱性磷酸酶(ALP)染色定量和实时定量聚合酶链式反应(qRT-PCR),探讨与特定浓度PD中JBMMSCs的成骨分化能力;采用ARS定量及qRT-PCR检测特定浓度以下PD对于JBMMSCs成骨分化能力的影响。结果 MTT试验发现0.1与0.5 μg/ml浓度PD不影响JBMMSCs生长(P>0.05),1.0、5.0、10.0 μg/ml浓度PD明显抑制JBMMSCs生长(P<0.05);ARS染色定量、ALP染色定量和qRT-PCR均发现与对照组比较,0.5和0.75 μg/ml浓度的PD组JBMMSCs成骨分化能力明显较低(P<0.05);ARS染色定量及qRT-PCR均显示0、0.1、0.2、0.3、0.4 μg/ml PD诱导下的JBMMSCs成骨分化能力相似(P>0.05)。结论 特定浓度以上PD可通过抑制JBMMSCs的成骨分化能力,引起骨坏死。

关键词: 帕米膦酸二钠, 间充质干细胞, 骨坏死, 成骨分化

Abstract: Objective To explore the cause of bisphosphate-related osteonecrosis by studying the effect of pamidronate disodium (PD) on osteogenic differentiation in jaw bone marrow mesenchymal stem cells (JBMMSCs). Methods JBMMSCs were isolated from the human jaw bone. The concentrations of PD suitable for the growth of JBMMSCs and subsequent studies were screened by thiazolyl blue (MTT) assay at concentrations of 0, 0.1, 0.5, 1.0, 5.0 and 10 μg/ml; The quantification of alizarin red S (ARS) staining and alkaline phosphatase (ALP) staining, and quantitative polymerase chain reaction (qRT-PCR) were used to compare the osteogenic ability of JBMMSCs at different concentrations of PD. Results MTT assay showed that 0.1 and 0.5 μg/ml PD did not affect the growth of JBMMSCs (P>0.05), while 1.0, 5.0 and 10 μg/ mL PD significantly inhibited the growth of JBMMSCs (P<0.05). The quantification of ARS staining and ALP staining, and qPCR showed that the osteogenic ability of JBMMSCs at 0.5 and 0.75 μg/ml of PD was significantly lower than that in the control group (P<0.05). ARS staining and qPCR showed similar osteogenic ability of JBMMSCs treated by 0, 0.1, 0.2, 0.3 or 0.4 μg/ml PD (P>0.05). Conclusion PD may induce osteonecrosis by inhibiting the osteogenic differentiation of JBMMSCs when over a threshold of dose.

Key words: Pamidronate disodium, Mesenchymal stem cells, Osteonecrosis, Osteogenic differentiation