聚3-羟基丁酸酯4-羟基丁酸酯/聚乙二醇/氧化石墨烯组织工程支架的制备和性能评价

Preparation and properties of poly3-hydroxybutyrate 4-hydroxybutyrate/polyethylene glycol/graphene oxide tissue-engineered scaffolds

背景:聚3-羟基丁酸酯4-羟基丁酸酯(poly3-hydroxybutyrate4-hydroxybutyrate,P34HB)支架因亲水性不足而在组织工程领域的应用受限。目的:探索P34HB、聚乙二醇(polyethylene glycol,PEG)共溶剂混合氧化石墨烯(graphene oxide,GO)复合静电纺丝支架的理化性能和生物相容性。方法:采用静电纺丝技术分别制备P34HB、P34HB/PEG、P34HB/PEG/GO支架材料,利用扫描电镜观察静电纺丝支架的三维结构,光学测量仪测定支架上水滴的静接触角,力学测试仪分析静电纺丝支架的拉伸应力应变情况和拉伸弹性模量。将3种支架分别与大鼠骨髓间充质干细胞共培养,MTT法检测细胞黏附,Alamar blue法检测细胞增殖,Live/Dead荧光染色观察细胞存活率,扫描电镜观察细胞形态。结果与结论:①扫描电镜显示3组支架纤维随机分布,P34HB、P34HB/PEG、P34HB/PEG/GO组纤维直径逐渐减少;②P34HB/PEG、P34HB/PEG/GO支架的接触角小于P34HB支架(P<0.01);③P34HB/PEG、P34HB/PEG/GO支架的应变率低于P34HB支架(P<0.05或P<0.01),P34HB/PEG/GO支架的弹性模量高于P34HB/PEG支架(P<0.05),3组支架抗拉强度比较差异无显著性意义(P>0.05);④P34HB/PEG/GO组共培养1,3,6 h的细胞黏附数量多于P34HB组、P34HB/PEG组(P<0.01),且P34HB/PEG组多于P34HB组(P<0.01);⑤P34HB/PEG/GO组共培养4,7 d的细胞增殖快于P34HB组、P34HB/PEG组(P<0.05或P<0.01);⑥共培养7 d后,扫描电镜显示P34HB/PEG/GO支架表面细胞量最大、铺展最好,Live/Dead荧光染色该组细胞存活率最高;⑦结果表明,P34HB/PEG/GO静电纺丝支架具有一定的力学性能与良好的亲水性,能有效促进细胞增殖、黏附。

BACKGROUND:The application of poly3-hydroxybutyrate 4-hydroxybutyrate(P34HB)scaffolds in tissue engineering has been limited due to lacking hydrophilicity.OBJECTIVE:To explore the physicochemical properties and biocompatibility of P34HB as well as polyethylene glycol(PEG)co-solvent mixed graphene oxide(GO)electroplating scaffolds.METHODS:The scaffolds of P34HB,P34HB/PEG and P34HB/PEG/GO had been prepared by electrospinning technology.The three-dimensional structure of the scaffold had been observed by scanning electron microscopy.The contact angle of water droplets on the scaffold had been measured by optical measuring instrument.The tensile stress,strain and elastic modulus of electrospinning frame were analyzed by mechanical tester.The three kinds of scaffolds had been cocultured with rat bone marrow mesenchymal stem cells respectively.Cell adhesion had been detected by MTT assay.Cell proliferation was detected by Alamar blue assay.Cell survival rate had been observed by Live/Dead fluorescence staining.Cell morphology had been observed by scanning electron microscopy.RESULTS AND CONCLUSION:(1)Scanning electron microscopy had showed random distribution of scaffold fibers in the three groups,and the fiber diameters of P34HB,P34HB/PEG,and P34HB/PEG/GO groups gradually decreased.(2)The contact angles of P34HB/PEG and P34HB/PEG/GO scaffolds were smaller than that of the P34HB scaffold(P<0.01).(3)The strain rates of P34HB/PEG and P34HB/PEG/GO scaffolds were lower than that of P34HB scaffold(P<0.05 or P<0.01),and the elastic modulus of P34HB/PEG/GO scaffolds was higher than that of P34HB/PEG scaffold(P<0.05).There was no significant difference in the tensile strength among the three groups(P>0.05).(4)At 1,3,and 6 hours of co-culture,the number of cell adhesion in the P34HB/PEG/GO group was higher than that in the P34HB and P34HB/PEG groups(P<0.01),and the number of cell adhesion in the P34HB/PEG group was higher than that in the P34HB group(P<0.01).(5)At 4 and 7 days of co-culture,the proliferation of cells in the P34HB/PEG/GO group was faster than that in the P34HB group and P34HB/PEG group(P<0.05 or P<0.01).(6)After 7 days of co-culture,scanning electron microscopy showed that the surface cell volume of P34HB/PEG/GO scaffold was largest and best spread,and the cell survival rate of Live/Dead fluorescence staining group was highest.(7)The results have showed that P34HB/PEG/GO electrospinning scaffold had certain mechanical properties and good hydrophilicity,and could effectively promote cell proliferation and adhesion.