中国实用口腔科杂志

中国实用口腔科杂志

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富血小板纤维蛋白对根尖牙乳头干细胞生物学性能的作用研究

付越，雷双，朱姝，刘尧，陈旭   

  1. 中国医科大学口腔医学院·附属口腔医院儿童口腔科，辽宁省口腔疾病临床医学研究中心，辽宁  沈阳  110002（第一作者现工作于大连医科大学口腔医学院口腔内科教研室）
  • 出版日期:2019-03-15 发布日期:2019-05-31
  • 基金资助:

    辽宁省高等学校基本科研项目（LQNK201723）；沈阳市科学技术计划项目（F16-205-1-25）

  • Online:2019-03-15 Published:2019-05-31

摘要：

目的　研究富血小板纤维蛋白（platelet rich fibrin，PRF）对根尖牙乳头干细胞（stem cells from apical papilla，SCAP）生物学性能的作用，探讨其应用于牙髓再生治疗（regenerative endodontic treatment，RET）、促进年轻恒牙牙根继续发育的机制。方法　原代分离培养SCAP，抽取静脉血获取PRF。实验按所用PRF的体积不同分为4组：1/8PRF组、1/2PRF组、1PRF组、0PRF组（对照组）。分别收集各组PRF上清液，制备条件培养基。通过MTT法检测PRF对SCAP增殖的影响；对经不同PRF条件培养基预处理的SCAP进行成骨诱导，应用Western Blot检测牙本质涎磷蛋白（dentin sialophosphoprotein，DSPP）、牙本质基质蛋白1（dentin matrix protein 1，DMP1）、骨钙素（osteocalcin，OCN）的表达水平，检测PRF对SCAP成牙本质或成骨分化的影响。结果　在条件培养基培养3 d和5 d时，与对照组相比，实验组PRF均显著促进SCAP的增殖（P < 0.05），3 d和5 d时，1/2PRF组促进SCAP增殖作用最强（P < 0.05）。PRF条件培养基预处理的SCAP经成骨诱导后，高表达DSPP和DMP1（P < 0.05）。结论　PRF可显著促进SCAP增殖和向成牙本质细胞分化，为PRF应用于RET奠定了生物学基础。

关键词: 富血小板纤维蛋白, 根尖牙乳头干细胞, 牙髓再生治疗

Abstract:

Objective　To study the effects of PRF on the biological properties of SCAP and to explore the underlying mechanism of PRF in promoting the root development in immature permanent tooth in regenerative endodontic treatment. Methods　SCAP was primary isolated. PRF was extracted from human venous blood. The experimental groups were divided according to the PRF volume：1/8PRF group，1/2PRF group，1PRF group and 0PRF group （control group）. The supernatant was collected and the conditioned medium was prepared. MTT assay was used to test the effects of PRF on the proliferation of SCAP. SCAP was pre-treated by PRF-conditioned medium for 3 days，and then cultured in osteogenic medium for 10 days. Western blot was used to detect the expression levels of DSPP，DMP1，and OCN in different groups in order to explore the effect of PRF on the odonto/osteogenic differentiation of SCAP. Results 　After being cultured in the conditioned medium for 3 days and 5 days， proliferation of SCAP was significantly promoted in all experimental groups compared with the control group（P < 0.05）.1/2PRF group was  more significant compared with the other two groups at 3 days and 5 days （P < 0.05）. Compared with the control group，the expression levels of DSPP and DMP1 were up-regulated after SCAP being pre-treated with PRF conditioned medium and osteogenic induction. Conclusion　PRF significantly stimulated the proliferation and odontogenesis of SCAP in vitro. The favorable biological effects of PRF on SCAP laid a theoretical foundation for the application of PRF in RET.

Key words: platelet rich fibrin, stem cells from apical papilla, regenerative endodontic treatment