载锌多壁碳纳米管/壳聚糖复合材料成骨性能的研究

Study on Osteogenic Properties of Multiwalled Carbon Nanotubes/chitosan Composite Containing Zinc

目的:探究载锌多壁碳纳米管/壳聚糖复合材料上的锌离子是否有促进成骨细胞增殖和分化的作用,以及该材料上锌离子的适宜浓度。方法:制备载有不同含量锌离子的多壁碳纳米管/壳聚糖复合材料,并用扫描电镜观察。其中,0.2%(质量百分数)锌含量的复合材料为低锌组,1%(质量百分数)锌含量的复合材料为中锌组,2.5%(质量百分数)锌含量的复合材料为高锌组,不含锌的复合材料为对照组。将MC3T3-E1细胞接种于4种材料表面进行培养,分别用MTT法、碱性磷酸酶试剂盒、ELISA法、激光共聚焦显微镜检测MC3T3-E1细胞在复合材料上的增殖量、碱性磷酸酶活性、骨钙素水平、细胞形态。结果:扫描电镜观察到该材料具有大量孔隙结构;mtt检测中24h、48h、72h时,中锌组细胞增值量高于对照组(P<0.05),低锌组细胞增值量与对照组无明显差距,高锌组细胞增值量小于对照组(P<0.05);碱性磷酸酶活性检测中24h、48h、72h时,中锌组高于对照组(P<0.05),低锌组与对照组无明显差距,高锌组小于对照组(P<0.05);骨钙素水平检测中48h时,中锌组骨钙素浓度高于对照组(P<0.05),低锌组骨钙素浓度与对照组无明显差距,高锌组骨钙素浓度小于对照组(P<0.05);激光共聚焦观察到,在细胞接种48h后,中锌组细胞伸展情况最好,低锌组细胞伸展情况较中锌组差,高锌组细胞伸展较差并且有部分细胞皱缩,对照组细胞伸展情况与低锌组无明显差异。结论:中锌组复合材料更有利于MC3T3-E1细胞的增殖和分化。

Objective：To explore whether zinc ions on the multiwalled carbon nanotubes/chitosan composite can promote proliferation and differentiation of osteoblast,and the appropriate concentration of zinc ion on the materials.Methods：The multiwalled carbon nanotubes/chitosan composites with different contents of zinc ions were prepared,and observed by scanning electron microscope.Thereinto,0.2%zinc content of the composite was low-zinc group,1% zinc content of the composite was medium-zinc group,2.5% zinc content of the composite was high-zinc group,and the composite without zinc was control group.The MC3T3-E1 cells were seeded on surface of the four kinds of composites.Using MTT methods,alkaline phosphatase assay kit,ELISA,and laser scanning confocal microscope,cell proliferation,alkaline phosphatase activity,osteocalcin level,and cell morphology were detected.Results：Scanning electron microscope picture showed that the composite material had a large number of pore structure.The proliferation of cells at 24 h,48hand 72 hin the medium-zinc group was higher than that in the control group（P〈0.05）,the low-zinc group was not significantly different from control group,and the high-zinc group was less than control group（P〈0.05）.Alkaline phosphatase activities at 24 h,48h,72 hin the medium-zinc group were higher than those in the control group（P〈0.05）,the low-zinc group was not significantly different from control group,and the high-zinc group was less than control group（P〈0.05）.Osteocalcin level at 48hin the medium-zinc group was higher than that in the control group（P〈0.05）,the low-zinc group was not significantly different from control group,and the high-zinc group was less than control group（P〈0.05）.Conclusion：The middle-zinc-group composites was more favorable to the proliferation and differentiation of MC3T3-E1cells.