摘要
目的:探讨牵张力(Stretch)对牙周膜成纤维细胞(periodontal ligament fibroblasts,PDLFs)成骨分化的影响及作用机制研究。方法:体外培养hPDLFs并将其分为对照组(Control组)、牵张力干预6 h组(Stretch 6 h组)、牵张力干预12 h组(Stretch 12 h组)、SDF-1/CXCR4通路激活剂组(CTCE-0214组)、牵张力干预12 h+SDF-1/CXCR4通路抑制剂组(Stretch 12 h+AMD3100组)。使用FlexCell系统对hPDLFs施加周期性牵张力。比色法和碱性磷酸酶(alkaline phosphatase,ALP)染色法检测hPDLFs ALP活性;茜素红染色观察钙化结节情况;RT-qPCR法检测与成骨相关基因表达水平;免疫组化法检测BSP、COL1A1阳性表达;Western blot检测SDF-1/CXCR4通路蛋白表达。结果:与Control组比较,Stretch 6 h组、Stretch 12 h组及CTCE-0214组hPDLFs ALP活性增强、钙化结节增多,RUNX2、OPN和OCN mRNA相对表达水平提高,BSP、COL1A1阳性表达及SDF-1/CXCR4通路蛋白表达量增加,且Stretch 12 h组hPDLFs的成骨分化能力高于Stretch 6 h组(P<0.05);与Stretch 12 h组比较,Stretch 12 h+AMD3100组hPDLFs ALP活性下降、钙化结节减少,RUNX2、OPN和OCN mRNA相对表达水平降低,BSP、COL1A1阳性表达及SDF-1/CXCR4通路蛋白表达量减少(P<0.05)。结论:施加周期性牵张力可通过激活SDF-1/CXCR4通路促进hPDLFs的成骨分化、加快正畸进展。
Objective:To investigate the effect and mechanism of stretch force(Stretch)on osteogenic differentiation of periodontal ligament fibroblasts(PDLFs).Methods:The hPDLFs were cultured in vitro and separated into Control group,stretch force intervention for 6 hours group(Stretch 6 h group),stretch force intervention for 12 hours group(Stretch 12 h group),SDF-1/CXCR4 pathway activator group(CTCE-0214 group),and stretch force intervention for 12 hours+SDF-1/CXCR4 pathway inhibitor group(Stretch 12 h+AMD3100 group).The FlexCell system was used to apply periodic stretch force to hPDLFs.Colorimetric and ALP staining methods were used to detect ALP activity in hPDLFs.Alizarin red staining was performed to observe the condition of calcified nodules.RT-qPCR method was used to detect the expression levels of osteogenic related genes.Immunohistochemistry was performed to detect positive expression of BSP and COL1A1.Western blot was performed to detect the SDF-1/CXCR4 pathway proteins.Results:Compared with the Control group,the Stretch 6 h group,Stretch 12 h group,and CTCE-0214 group showed enhanced ALP activity and increased calcified nodules in hPDLFs.The relative expression levels of RUNX2,OPN,and OCN mRNA were increased,and the positive expression of BSP,COL1A1,and the expression of SDF-1/CXCR4 pathway proteins were also increased.Moreover,the osteogenic differentiation ability of hPDLFs in the Stretch 12 h group was higher than that in the Stretch 6 h group(P<0.05).Compared with the Stretch 12 h group,the Stretch 12 h+AMD3100 group showed a decrease in hPDLFs ALP activity and a reduction in calcified nodules.The relative expression levels of RUNX2,OPN,and OCN mRNA were decreased,and the positive expression of BSP,COL1A1,and the expression of SDF-1/CXCR4 pathway proteins were also decreased(P<0.05).Conclusion:Applying periodic stretch force can promote osteogenic differentiation of hPDLFs and accelerate orthodontic progression by activating the SDF-1/CXCR4 pathway.
作者
张帅
韩桂贤
于向前
马青
黄亭
ZHANG Shuai;HAN Gui-xian;YU Xiang-qian;MA Qing;HUANG Ting(Department of Stomatology,The Second Affiliated Hospital of Hebei North University,Hebei Zhangjiakou 075100,China;Department of Stomatology,Xuanhua District Hospital of Zhangjiakou City,Hebei Zhangjiakou 075100,China;Department of Clinical Laboratory,The Seventh People's Hospital of Hebei Province,Hebei Dingzhou 073000,China;Department of Stomatology,Cangzhou People's Hospital,Hebei Cangzhou 061000,China)
出处
《临床口腔医学杂志》
2025年第11期647-652,共6页
Journal of Clinical Stomatology
基金
2025年度河北省医学科学研究课题(20250907)
张家口市重点研发计划项目(2421101D)。
