南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (9): 1304-1309.doi: 10.12122/j.issn.1673-4254.2021.09.03

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神经生长因子联合牙髓干细胞可促进大鼠种植体周围骨结合

路晓淼，田瑞雪，刘姗姗，徐锦程   

  1. 蚌埠医学院第一附属医院口腔科，安徽 蚌埠 233004
  • 出版日期:2021-09-20 发布日期:2021-09-30

Nerve growth factor combined with dental pulp stem cells promotes peri-implant osseointegration in rats

LU Xiaomiao, TIAN Ruixue, LIU Shanshan, XU Jincheng   

  1. Department of Stomatology, First Affiliated Hospital of Bengbu Medical College, Bengbu 233004, China
  • Online:2021-09-20 Published:2021-09-30

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摘要： 目的 研究神经生长因子（NGF）联合牙髓干细胞（DPSCs）调节种植体周围骨结合的作用及机制。方法 培养SD大鼠的DPSCs并随机分3组，每组实验重复4次。对照组：不含药物的培养基处理；NFG组：含有NGF的培养基处理；NFG+K252a组：含有NGF及TrkA拮抗剂+K252a的培养基处理。成骨诱导后检测钙结节，Western blot检测RUNT相关转录因子2（RUNX2）、骨钙素（OCN）的表达量。SD大鼠建立股骨种植模型并分为对照组、DPSCs组、DPSCs+NGF组、DPSCs+K252a组、DPSCs+NGF+K252a组，8只/组。干预4周后进行种植体周围骨组织的micro-CT检查及HE染色并采用Western blot检测RUNX2、OCN表达量。结果 NGF组DPSCs的钙结节数目及RUNX2、OCN的表达水平均高于对照组（P<0.05）；NGF+K252a组DPSCs的钙结节数目及RUNX2、OCN的表达水平均低于NGF组（P<0.05）。DPSCs组及DPSCs+NGF组大鼠种植体周围骨组织的骨矿密度（BMD）、骨小梁厚度（Tb.Th）、骨小梁数量（Tb.N）及RUNX2、OCN的表达水平均明显高于对照组（P<0.05）且DPSCs+NGF组种植体周围骨组织的BMD、Tb.Th、Tb.N及RUNX2、OCN的表达水平高于DPSCs组（P<0.05）；DPSCs+K252a组种植体周围骨组织的BMD、Tb.Th、Tb.N及RUNX2、OCN的表达水平与DPSCs组无显著差异（P>0.05）；DPSCs+NGF+K252a组种植体周围骨组织的BMD、Tb.Th、Tb.N及RUNX2、OCN的表达水平均明显低于DPSCs+NGF组（P<0.05）。结论 NGF联合DPSCs能够促进大鼠种植体周围骨结合，NGF通过TrkA促进DPSCs成骨分化是可能的分子机制。

关键词: 牙髓干细胞；神经生长因子；种植体；骨结合；成骨分化

Abstract: Objective To observe the effect of nerve growth factor (NGF) combined with dental pulp stem cells (DPSCs) on peri-implant osseointegration in rats and explore the underlying mechanism. Methods Cultured DPSCs were treated with NFG or with NFG plus the TrkA antagonist K252a, and after osteogenic induction, the formation of calcium nodules was observed andthe expressions of RUNX2 and osteocalcin (OCN) were detected with Western blotting. SD rat models with femoral implantation of dental implants were established and divided into control, DPSCs, DPSCs+NGF, DPSCs+K252a, and DPSCs+NGF+K252a groups, and in all but the control group, DPSCs were injected locally before placement of the implants (n=8); NGF, K252a and their combination were injected intraperitoneally on a daily basis for 4 weeks. All the rats underwent micro-CT, and the peri-implant bone tissues were collected for HE staining and detection of RUNX2 and OCN expressions using Western blotting. Results In cultured DPSCs, the number of calcium nodules and the expression levels of RUNX2 and OCN were significantly higher in NGF group than in the control group (P<0.05), and were significantly lower in NGF+K252a group than in NGF group (P<0.05). In the rat models, the BMD, TB. Th, TB. N and expressions of RUNX2 and OCN in the peri-implant bone tissues were significantly higher in DPSCs group and DPSCs+NGF group than in the control group (P<0.05), significantly higher in DPSCs+NGF group than in DPSCs group (P<0.05), comparable between DPSCs+K252a group and DPSCs group (P> 0.05), and significantly lower in DPSCs+NGF+K252a group than in DPSCs+NGF group (P<0.05). Conclusion NGF combined with DPSCs can promote osseointegration in the peri-implant tissues in rats possibly due to a TrkA-mediated effect of NGF for promoting osteogenic differentiation of the DPSCs.

Key words: dental pulp stem cells; nerve growth factor; implant; osseointegration; osteogenic differentiation