中国组织工程研究

中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (47): 7591-7596.doi: 10.3969/j.issn.2095-4344.2014.47.009

• 组织工程骨及软骨材料 tissue-engineered bone and cartilage materials • 上一篇    下一篇

磷离子对三维培养下人骨髓间充质干细胞的影响

雷  群，陈  江，黄文秀，吴  东，林  东   

  1. 福建医科大学附属口腔医院种植科，福建省福州市  350000
  • 修回日期:2014-10-18 出版日期:2014-11-19 发布日期:2014-11-19
  • 通讯作者: 陈江，博士，教授，主任医师，博士生导师，福建医科大学附属口腔医院种植科，福建省福州市 350000
  • 作者简介:雷群，女，1982年生，湖北省人，汉族，2013年福建医科大学毕业，博士，医师，主要从事口腔材料和组织工程学研究。
  • 基金资助:

    2012年福建省临床重点专科项目资助(2012)1589；福建省科技厅重点项目(2013Y0039)

Effect of phosphorus ions on human bone marrow mesenchymal stem cells under three-dimensional culture 

Lei Qun, Chen Jiang, Huang Wen-xiu, Wu Dong, Lin Dong   

  1. Department of Dental Implant, Stomatological Hospital of Fujian Medical University, Fuzhou 350000, Fujian Province, China
  • Revised:2014-10-18 Online:2014-11-19 Published:2014-11-19
  • Contact: Chen Jiang, M.D., Professor, Chief physician, Doctoral supervisor, Department of Dental Implant, Stomatological Hospital of Fujian Medical University, Fuzhou 350000, Fujian Province, China
  • About author:Lei Qun, M.D., Physician, Department of Dental Implant, Stomatological Hospital of Fujian Medical University, Fuzhou 350000, Fujian Province, China
  • Supported by:

    the Key Clinical Specialty Discipline Construction Program of Fujian Province, No. (2012)1589; Key Project of Fujian Provincial Department of Science and Technology, No. 2013Y0039

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摘要：

背景：以往研究是关于含磷复合物对动物源性或人源性其他干细胞的影响，三维情况下磷离子对人骨髓间充质干细胞作用研究尚未见。
目的：探讨磷离子对三维培养条件下人骨髓间充质干细胞的影响。
方法：实验分为6组，将人骨髓间充质干细胞接种在三维聚苯乙烯培养支架上，细胞分别在无血清生长培养基和添加4，8 mmol/L磷离子的无血清生长培养基中培养21 d；在二维聚苯乙烯培养板上培养的细胞作为相应的对照组。CCK8法检测细胞增殖情况，RT-PCR检测成骨分化标记性基因的表达，茜素红染色检测人骨髓间充质干细胞成骨分化生成的矿化结节。
结果与结论：培养4，7，14，21 d，相对于二维的细胞培养，磷离子的促细胞生长作用在三维聚苯乙烯培养支架上表现更明显(P < 0.05)。相对与三维培养对照组，培养7，14 d时，胶原蛋白Ⅰ基因的表达在4，8 mmol/L 磷离子三维培养组显著增加(P < 0.05)；培养14 d时，碱性磷酸酶基因的表达在4，8 mmol/L磷离子三维培养组显著增加(P < 0.05)；培养14，21 d时，骨钙素基因的表达在4，8 mmol/L磷离子三维培养组显著增加(P < 0.05)。培养21 d时，在4，8 mmol/L磷离子三维培养组可生成矿化结节，在三维培养对照组无矿化结节生成。结果说明，磷离子可以促进人骨髓间充质干细胞的增殖和成骨分化；相对于二维的细胞培养，磷离子的促生长作用在三维聚苯乙烯培养支架上表现更明显。


中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程


全文链接：

关键词: 生物材料, 骨生物材料, 磷, 二维培养, 三维培养, 支架, 干细胞, 培养基, 诱导成骨, 细胞增殖, 骨替代材料, 骨移植材料

Abstract:

BACKGROUND: Previous researches have focused on the effect of phosphorus compounds on stem cells from animals or from human. But there is no study on the effect of phosphorus ions on human bone marrow mesenchymal stem cells under three-dimensional culture.
OBJECTIVE: To explore the effect of phosphorus ions on human bone marrow mesenchymal stem cells under three-dimensional culture.
METHODS: There were six groups in the experiment. Human bone marrow mesenchymal stem cells were inoculated in three-dimensional polystyrene scaffolds and then subjected to serum-free growth medium (group 3-GM) or serum-free growth medium containing 4 mmol/L (group 3-4P), 8 mmol/L (group 3-8P) phosphorus ions for 21 days, respectively. Cells cultured on the two-dimensional polystyrene scaffolds were used as control groups (groups 2-GM, 2-4P, 2-8P). Cellular proliferation was examined by cell counting kit-8; the mRNA expressions of osteogenic marker genes were assessed by RT-PCR; the formation of mineralized nodules for the osteogenic differentiation of human bone marrow mesenchymal stem cells was examined by Alizarin red staining.
RESULTS AND CONCLUSION: Compared with the two-dimensional culture, the growth of human bone marrow mesenchymal stem cells induced by phosphorus ions were more obvious in the three-dimensional polystyrenes scaffolds at days 4, 7, 14 and 21 (P < 0.05). Compared with the group 3-GM, the mRNA expression of collagen type I in groups 3-4P and 3-8P increased more significantly at days 7 and 14 (P < 0.05); the mRNA expression of alkaline phosphatase in groups 3-4P and 3-8P increased more significantly at day 14 (P < 0.05); the mRNA expression of osteocalcin in groups 3-4P and 3-8P increased more significantly at days 14 and 21 (P < 0.05). Mineralized nodules were formed in groups 3-4P and 3-8P but not in group 3-GM at day 21. So we concluded that phosphorus ions can promote proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells in three-dimensional polystyrenes scaffolds. Compared with the two-dimensional cell culture, the promoting growth effect of phosphorus ions on human bone marrow mesenchymal stem cells in three dimensional polystyrenes scaffolds are more obvious.


中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程


全文链接：

Key words: tissue engineering, phosphorus, stem cells

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