PEGS/β-TCP屏障膜在促进大鼠颅骨缺损骨组织再生中的作用

Application of PEGS/β-TCP membrane in promoting bone regeneration of rat calvarial defect

目的:通过体内和体外实验,探讨PEGS/β-TCP复合膜诱导骨组织再生的效果。方法:将PEGS/β-TCP复合膜与大鼠骨髓间充质干细胞(rBMSCs)共培养,观察rBMSCs细胞黏附、增殖及成骨分化情况。建立大鼠颅骨缺损动物模型,将PEGS/β-TCP复合膜植入缺损特定区域,通过Micro-CT观察骨缺损区域新骨形成情况;再通过脱钙组织H-E染色,观察炎性浸润程度和新骨形成的情况。采用SPSS 22.0软件包对数据进行统计学分析。结果:rBMSCs可良好黏附和广泛分布于PEGS/β-TCP复合膜上。rBMSCs接种1、4和7 d后进行的dsDNA定量分析结果显示,从第1天到第7天,细胞DNA含量逐渐增加;培养7 d后,实验组细胞DNA含量显著高于对照组(P<0.001)。对接种于PEGS/β-TCP复合膜上的rBMSCs进行体外成骨诱导,其碱性磷酸酶(ALP)染色浓度显著增高。ALP活性半定量检测显示,实验组较对照组细胞ALP活性增加更为显著(P<0.01)。成骨诱导21 d后,实验组与对照组茜素红矿化染色结果与ALP染色结果一致。动物模型Micro-CT检测及组织H-E染色均显示,实验组各检测时间点的新骨形成量显著高于对照组,且各组炎症反应均轻微可控。结论:PEGS/β-TCP复合膜具有理想的细胞相容性和骨引导再生效果。

PURPOSE:The purpose of this study was to explore the effect of PEGS/β-TCP membrane on guided bone regeneration.METHODS:PEGS/β-TCP membrane was co-cultured with rat bone marrow mesenchymal stem cells(rBMSCs)to observe cell adhesion,proliferation and osteogenic differentiation.Animal model of rat skull defect was established,and PEGS/β-TCP membrane was implanted in the area of the defect.Micro-CT was used to observe the formation of new bone in bone defect area,and H-E staining of decalcified tissue was used to observe the degree of inflammatory infiltration and new bone formation.Statistical analysis was performed with SPSS 22.0 software package.RESULTS:rBMSCs can be well adhered and widely distributed on PEGS/β-TCP membrane.Quantitative analysis of dsDNA at 1,4 and 7 days showed that the DNA content in both groups increased from day 1 to day 7,and there was significant difference between the control group and the experimental group at day 7(P<0.001).Alkaline phosphatase(ALP)activity of rBMSCs on PEGS/β-TCP membrane increased significantly after osteogenic induction in vitro.ALP activity of the experimental group was significantly higher than that of the control group(P<0.01).The results of Alizarin red mineralization staining and ALP staining showed the same trend at day 21.Micro-CT examination and H-E staining showed that the formation of new bone in the experimental group was significantly higher than that in the control group,and the inflammatory reaction in each group was mild.CONCLUSIONS:This study shows that PEGS/β-TCP composite membrane had ideal cytocompatibility and guided bone regeneration effect,which provides theoretical support for further study and application of PEGS/β-TCP membrane in guided bone regeneration.