实用医学杂志 ›› 2021, Vol. 37 ›› Issue (10): 1257-1262.doi: 10.3969/j.issn.1006⁃5725.2021.10.005

• 基础研究 • 上一篇    下一篇

电活性生物膜通过调控巨噬细胞极化促进骨再生修复的体外研究 

朱培君, 赖春花, 程鸣威 ,何逸恒, 徐淑兰   

  1. 南方医科大学口腔医院种植中心(广州510300)
  • 出版日期:2021-05-25 发布日期:2021-05-25
  • 通讯作者: 徐淑兰 E⁃mail:xushulan@vip.163.com
  • 基金资助:
    南方医科大学科研培育计划项目⁃面上项目(编 号:PY2020011);广东省医学科研基金(编号:A2018485);广东省 中医药局科研项目(编号:20211274);南方医科大学科研启动计 划青年培育项目(编号:PY2017N012) 通信作者:徐淑兰 E⁃mail:xushulan@vip.163.com10.3969/j.issn.1006⁃5725.2021.10.005 基金项目:南方医科大学科研培育计划项目⁃面上项目(编 号:PY2020011);广东省医学科研基金(编号:A2018485);广东省 中医药局科研项目(编号:20211274);南方医科大学科研启动计 划青年培育项目(编号:PY2017N012) 

Promotive role of electrically biomembranes in bone regeneration through regulating the polarization of Macrophages 

ZHU Peijun,LAI Chunhua,CHENG Mingwei,HE Yiheng,XU Shulan.    

  1. Center of Oral Implantolo⁃ gy,Stomatological Hospital,Southern Medical University,Guangzhou 510300,China 

  • Online:2021-05-25 Published:2021-05-25
  • Contact: XU Shulan E⁃mail:xushulan@vip.163.com

摘要:

目的 为了研发一种具骨免疫调节性能的屏障膜,本研究制备电活性生物膜并明确其对 巨噬细胞极化分型的调控作用,再进一步探究电活性生物膜介导的巨噬细胞极化对成骨分化的影响。方法 制备带电的聚偏氟乙烯⁃三氟乙烯[Poly(vinylidene fluoride trifluoroethylene),P(VDF⁃TrFE)]薄膜。构建 P(VDF⁃TrFE)膜、巨噬细胞、大鼠骨髓间充质干细胞(rat bone marrow mesenchymal stem cells,rBMSCs 共培养体系,不带电 P(VDF⁃TrFE)膜作为对照组,应用流式细胞术、PCR 技术检测带电 P(VDF⁃TrFE)膜上巨噬细胞的极化表型。在膜⁃巨噬细胞条件培养基的刺激下,应用碱性磷酸酶(alkaline phosphatase,ALP)、茜素红染色等技术检测 BMSCs 的成骨活性。结果 带电 P(VDF⁃TrFE)膜具良好的电学稳定性。巨噬细胞在带电膜上表现为 M2 型特征蛋白 CD206 的高表达,PCR 结果也显示了 Arg⁃1、IL⁃10 在带电膜上有较高的表达(P<0.05)。带电薄膜⁃巨噬细胞环境下,rBMSCs 分泌的 ALP 及矿化结节较对照组多。结论 电活性 P(VDF⁃TrFE)膜可有效促进巨噬细胞向 M2 表型极化,在其介导的巨噬细胞环境下,rBMSC 显示出较高的成骨活性,这对骨缺损区的再生修复具有重要意义。

目的 为了研发一种具骨免疫调节性能的屏障膜,本研究制备电活性生物膜并明确其对 巨噬细胞极化分型的调控作用,再进一步探究电活性生物膜介导的巨噬细胞极化对成骨分化的影响。方法 制备带电的聚偏氟乙烯⁃三氟乙烯[Poly(vinylidene fluoride trifluoroethylene),P(VDF⁃TrFE)]薄膜。构建 P(VDF⁃TrFE)膜、巨噬细胞、大鼠骨髓间充质干细胞(rat bone marrow mesenchymal stem cells,rBMSCs 共培养体系,不带电 P(VDF⁃TrFE)膜作为对照组,应用流式细胞术、PCR 技术检测带电 P(VDF⁃TrFE)膜上巨噬细胞的极化表型。在膜⁃巨噬细胞条件培养基的刺激下,应用碱性磷酸酶(alkaline phosphatase,ALP)、茜素红染色等技术检测 BMSCs 的成骨活性。结果 带电 P(VDF⁃TrFE)膜具良好的电学稳定性。巨噬细胞在带电膜上表现为 M2 型特征蛋白 CD206 的高表达,PCR 结果也显示了 Arg⁃1、IL⁃10 在带电膜上有较高的表达(P<0.05)。带电薄膜⁃巨噬细胞环境下,rBMSCs 分泌的 ALP 及矿化结节较对照组多。结论 电活性 P(VDF⁃TrFE)膜可有效促进巨噬细胞向 M2 表型极化,在其介导的巨噬细胞环境下,rBMSC 显示出较高的成骨活性,这对骨缺损区的再生修复具有重要意义。

关键词:

生物材料, 巨噬细胞, 骨免疫, 成骨分化, 免疫调节性能

Abstract:

Objective For the purpose of developing a barrier membrane with osteoimmunomodulatory property,this study prepared electroactive biomembranes and clarified its regulatory effects on macrophage polar⁃ ization typing,and further explored the effect of electroactive biomembrane⁃mediated macrophage polarization on osteogenic differentiation. Methods Preparing charged poly(vinylidene fluoride trifluoroethylene)[P(VDF⁃TrFE)] membrane. A co⁃culture system of P(VDF⁃TrFE)membrane,macrophages,and bone marrow mesenchymal stem cells(BMSCs)was constructed. Uncharged P(VDF ⁃TrFE)membrane was set as a control group. Flow cytometry and PCR techniques were used to detect the phenotype of macrophages on the charged P(VDF⁃TrFE)membrane. Alkaline phosphatase(ALP)and alizarin red staining techniques were used to detect the osteogenic activity of BM⁃ SCs under the stimulation of P(VDF ⁃ TrFE)membrane ⁃ macrophage conditioned medium. Results Charged P (VDF⁃TrFE)membrane had good electrical stability. Macrophages showed high expression of M2 characteristic protein CD206 on the charged membrane,PCR results also showed that Arg⁃1 and IL⁃10 had higher expression on the charged membrane(P<0.05),and BMSCs produced more ALP and mineralized nodules in the charged membrane⁃macrophage environment. Conclusion The electroactive P(VDF⁃TrFE)membrane can effectively pro⁃ mote the polarization of macrophages to the M2 phenotype. Under its mediated immune environment,BMSCs show high osteogenic activity,which is of great significance to the regeneration and repair of bone defects. 

Key words:

biological materials, macrophages, osteoimmunology, osteogenic differentiation, im?munomodulatory property