NSAIDs对钛片表面成骨细胞增殖及功能活性的影响

Effect of non-steroidal anti-inflammatory drugs on growth and functional activity of osteoblasts in vitro

目的通过建立种植体表面成骨模型,研究新型与传统非甾体类抗炎药(nonsteroidal antiinflammatory drugs,NSAIDs)对钛片表面MG-63成骨细胞增殖、形态、粘附、活性和成骨相关基因表达的影响。方法将MG-63成骨细胞接种至大颗粒喷砂酸蚀(sand-blasted,largegrit,acid-etched,SLA)钛片表面,建立种植体表面与成骨细胞结合的模型。设置一氧化氮供体型氟比洛芬(NO-氟比洛芬)组、氟比洛芬组与无药物对照组。CCK-8检测细胞增殖能力,扫描电镜观察细胞形态,MTT检测细胞粘附能力,化学法检测碱性磷酸酶(ALP)活性,茜素红染色观察钙化结节,RT-qPCR检测各组成骨细胞ALP、OCN、Runx-2 mRNA表达情况。结果细胞增殖检测结果显示,不同药物浓度组间的细胞增殖数量有统计学差异(P<0.01);NO-氟比洛芬组和氟比洛芬组细胞增殖数量均高于无药物对照组,且NO-氟比洛芬组高于氟比洛芬组,差异均有统计学意义(P<0.01)。扫描电镜观察结果显示,NO-氟比洛芬组成骨细胞最先开始复层生长。细胞粘附检测结果显示,NO-氟比洛芬组细胞粘附数量低于氟比洛芬组和无药物对照组(P<0.01)。ALP活性检测结果显示,NO-氟比洛芬组和氟比洛芬组ALP活性低于无药物对照组(P<0.01)。茜素红染色结果显示,NO-氟比洛芬组及氟比洛芬组实验观察期内未观察到典型的红色深染钙化结节。RT-qPCR分析显示,NO-氟比洛芬组OCN mRNA和氟比洛芬组Runx-2 mRNA表达量高于无药物对照组,ALP mRNA表达量低于无药物对照组(P<0.01)。结论NO-氟比洛芬和氟比洛芬均有效促进钛片表面成骨细胞的增殖能力和部分成骨相关基因表达,NO-氟比洛芬的促进效果更为明显。

Objective To investigate the effects of new and traditional non-steroidal anti-inflammatory drugs(NSAIDs)on the proliferation,morphology,adhesion,activity and expression of osteoblast related genes of MG-63 osteoblasts by establishing a osteogenic model of implants.Methods MG-63 cells were implanted on sand-blasted,largegrit,acid-etched titanium plates surface(SLA)to establish a model of osseointegration between implant surface and osteoblasts.The experiments included NO-flurbiprofen group,flurbiprofen group and the control group.Cell proliferation was detected by CCK-8 assay,cell morphology was observed by scanning electron microscopy(SEM),cell adhesion was detected by MTT assay,alkaline phosphatase(ALP)activity was detected by chemical method,calcified nodules was observed after Alizarin red staining,and the expression of bone formation related genes ALP,OCN,Runx-2 was detected by RT-qPCR.Results CCK-8 assay showed that statistical difference was observed in cell proliferation among the groups treated with different drug concentrations(P<0.01).Cell proliferation was higher in the NO-flurbiprofen group and the flurbiprofen group than the control group,and in the NO-flurbiprofen group than the flurbiprofen group(P<0.01).SEM displayed that the cells from the NO-flurbiprofen group grew to multiple lays first.Cell adhesion results showed that the number of cell adhesion was lower in the NO-flurbiprofen group than the flurbiprofen group and the control group(P<0.01).ALP activity of drug groups was lower than the control group(P<0.01).No typical red deeply stained calcified nodules were observed in neither NO-flurbiprofen group nor flurbiprofen group during the observation period.RT-qPCR indicated that the mRNA expression of OCN in the NO-flurbiprofen group and Runx-2 in the flurbiprofen group was higher than those of the control group,while the mRNA level of ALP was lower than that of the control group(P<0.01).Conclusion Both NO-flurbiprofen and flurbiprofen effectively promote the proliferation of osteoblasts on the surface of titanium plates and the expression of some osteogenic related genes.The promotion effect of NO-flurbiprofen is more significant.