中国组织工程研究

中国组织工程研究 ›› 2015, Vol. 19 ›› Issue (3): 399-404.doi: 10.3969/j.issn.2095-4344.2015.03.013

• 细胞外基质材料 extracellular matrix materials • 上一篇    下一篇

不同质量浓度釉基质蛋白培养人牙周膜细胞的生物活性

曲 哲1，张静莹2，郭 英1，马卫东1，马 岚1   

  1. 1大连市口腔医院种植科，辽宁省大连市 116021；2大连大学医学部，辽宁省大连市 116622
  • 出版日期:2015-01-15 发布日期:2015-01-15
  • 通讯作者: 曲哲，大连市口腔医院种植科，辽宁省大连市 116021
  • 作者简介:曲哲，男，1977年生，吉林省长春市人，2008年维也纳医科大学毕业，博士，副教授，主要从事种植体表面改性和骨组织工程研究。
  • 基金资助:

    人力资源和社会保障部归国人员启动基金(2011508)；辽宁省教育厅高校科研项目(2008027)；大连市民生科技项目(2013E15SF169)

Influence of different concentrations of enamel matrix proteins on bioactivity of human periodontal ligament cells

Qu Zhe1, Zhang Jing-ying2, Guo Ying1, Ma Wei-dong1, Ma Lan1   

  1. 1Department of Dental Implantation, Dalian Stomatological Hospital, Dalian 116021, Liaoning Province, China; 2Medicine Department, Dalian University, Dalian 116622, Liaoning Province, China
  • Online:2015-01-15 Published:2015-01-15
  • Contact: Qu Zhe, Department of Dental Implantation, Dalian Stomatological Hospital, Dalian 116021, Liaoning Province, China
  • About author:Qu Zhe, M.D., Associate professor, Department of Dental Implantation, Dalian Stomatological Hospital, Dalian 116021, Liaoning Province, China
  • Supported by:

    the Start Fund for Returnees of Ministry of Human Resources and Social Security of China, No. 2011508; the Scientific Research Project in High Education of Liaoning Provincial Education Department, No. 2008027; the Science and Technology Project for People’s Livelihood in Dalian, No. 2013E15SF169

PDF

521

被引次数

2

摘要：

背景：大量研究证实釉基质蛋白可促进成骨细胞和成牙骨质细胞的再生，将其运用于牙周缺损治疗可达到接近生理性的牙周再生。

 

目的：观察不同质量浓度釉基质蛋白对人牙周膜细胞增生、分化和迁移的影响。

 

方法：取第3代人牙周膜细胞，以含不同质量浓度釉基质蛋白(0，12.5，25，50，100，250 mg/L)的无血清DMEM培养基培养。培养24 h后，采用3H-胸腺嘧啶核苷掺入法检测细胞增殖，MTT法检测细胞活性；培养48 h后，检测细胞碱性磷酸酶活性及骨钙素分泌；待细胞融合为单层，去除细胞培养液，以移液管头将单层细胞制备出1 mm宽的细胞切口，持续24 h观察细胞融合情况。

 

结果与结论：当釉基质蛋白质量浓度在0-100 mg/L范围内，随着其质量浓度的升高，细胞增殖、活性、碱性磷酸酶活性、骨钙素分泌均逐渐升高，以100 mg/L升高最明显；当釉基质蛋白质量浓度增至250 mg/L时，细胞增殖、活性、碱性磷酸酶活性、骨钙素分泌均有所下降，但仍高于0 mg/L组。100 mg/L组在初始观察      6 h时，创缘周围的细胞开始向中心生长，待培养12 h时，创缘两侧细胞开始融合，培养20 h后创缘两侧细胞融合完全创缘完全关闭完全，创面愈合优于其他质量浓度组。结果表明釉基质蛋白具有促进牙周膜细胞增殖、分化与迁移的能力。

 

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, 口腔生物材料, 釉基质蛋白, 牙周膜细胞, 细胞迁移, 碱性磷酸酶, 骨钙素

Abstract:

BACKGROUND: Numerous studies have confirmed that enamel matrix proteins can promote the regeneration of osteoblasts and cementoblast, and then it can achieve approaching physiological periodontal regeneration in the treatment of periodontal defects.

OBJECTIVE: To observe the effects of different concentrations of enamel matrix proteins on proliferation, viability, differentiation and migration of human periodontal ligament cells.
METHODS: The human periodontal ligament cells at the third generation were gained, and then cultured in serum-free DMEM containing different concentrations of enamel matrix proteins (0, 12.5, 25, 50, 100, 250 mg/L). After 24 hours of culture, proliferation and viability of periodontal ligament cells were measured using [(3)H]-thymidine uptake and MTT assay. After 48 hours of culture, alkaline phosphatase activity and osteocalcin production were detected with commercial available test kits. When the cells grew as a monolayer, the cell culture fluid was removed, and then with a pipette head, a cell incision, 1 mm wideness, was prepared in a monolayer of cells to further observe the cell fusion continuously within 24 hours.
RESULTS AND CONCLUSION: The proliferation, viability and differentiation of periodontal ligament cells were gradually increased with the concentration increase of enamel matrix proteins (0-100 mg/L). When the concentration of enamel matrix proteins was 250 mg/L, these parameters began to decrease, but the levels were still higher than those in the 0 mg/L enamel matrix protein group. In the 100 mg/L group, the cells in the wound edge began to grow towards the center in the initial 6 hours, then became confluent at 12 hours, and until the 20th hour of culture, the cells in the two sides of the wound edge were completely fused to fully close the wound edge, indicating a better wound healing in the     100 mg/L group than the other groups. These findings suggest that enamel matrix proteins can stimulate proliferation, viability, differentiation and migration of periodontal ligament cells.

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: Periodontal Ligament, Extracellular Matrix Proteins, Cell Movement

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