载银锌介孔钙硅纳米粒子与聚己内酯复合材料的体外生物活性研究

In vitro biological activity study on the material composited with nanosilver and nanozinc incorporated mespoporous calcium-silicate and PCL

目的评价载银(Ag)或锌(Zn)的介孔钙硅纳米粒子(mesoporous calcium-silicate nanoparticles,MCSNs)和聚己内酯(poly-ε-caprolactone,PCL)的复合材料的理化性能、体外生物活性及促进成骨能力。方法通过加热熔融法制备PCL-MCSNs、PCL-Zn-MCSNs、PCL-Ag-MCSNs和PCL-Ag-Zn-MCSNs(Ag与Zn物质的量比为1∶1),表征其表面形貌、机械性能和体外降解性能。复合材料与根尖乳头干细胞(stem cell from the apical papilla,SCAP)共培养,观察其表面的细胞附着和羟磷灰石晶体(hydroxyapatite,HA)形成情况。通过CCK-8法、碱性磷酸酶(ALP)活性测定和茜素红染色,评价材料的细胞毒性和促进体外成骨效果。结果所有复合材料的表面平整均匀,PCL-MCSNs的压缩强度最高,为3.05 MPa。PCL-Zn-MCSNs的表面HA形成最多、降解最快、表面附着的细胞密度最高,细胞粘附性最好。除PCL-Ag-MCSNs和PCL-Ag-Zn-MCSNs外,其他实验组都能促进细胞增殖,ALP活性升高,产生的钙化结节含量均高于PCL,其中PCL-Zn-MCSNs显著高于其他组(P<0.05)。结论PCL和载银或锌的MCSNs的复合材料具有良好的机械性能和降解性能,但细胞粘附性、细胞增殖性、ALP活性和钙化结节形成能力随Zn含量增加而增强,提示含Zn的该类型复合材料的体外生物活性良好,在促成牙/成骨方面有着优越的潜能,有望成为新型的口腔颌面骨缺损的修复材料。

Objective To investigate the physical and chemical properties,biological activity in vitro and pro-osteogenic properties of poly-ε-caprolactone(PCL)and mesoporous calcium silicate nanoparticles(MCSNs)containing silver or zinc composite materials.Methods The composite materials of PCL-MCSNs,PCL-Zn-MCSNs,PCL-Ag-MCSNs,PCL-Ag-Zn-MCSNs were prepared by heating and melting method and characterized to test their surface morphology,mechanical properties and degradation properties in vitro.The composite materials were co-cultured with stem cells from the apical papilla(SCAP)to observe cell adhesion and hydroxyapatite(HA)formation on the surface.The cytotoxicity of composite materials and pro-osteogenic properties in vitro were studied by CCK-8 method,alkaline phosphatase(ALP)activity determination and Alizarin Red staining.Results The surface of each group was smooth and even.The compressive strength of the PCL-MCSNs group was 3.05 MPa,which was the highest.PCL-Zn-MCSNs showed the most HA formation,the fastest degradation,the highest surface cell density and the best cell adhesion.Except for PCL-Ag-MCSNs and PCL-Ag-Zn-MCSNs,all the experimental groups could promote cell proliferation,increase ALP activity,and produce more calcified nodular content than PCL,among which PCL-Zn-MCSNs was significantly higher than the other groups(P<0.05).Conclusion The composite materials of PCL and MCSNs containing silver or zinc reveal good mechanical and degradation properties.The cell adhesion,cell proliferation,ALP activity and calcified nodule formation ability are enhanced with the increase of Zn content,suggesting that this kind of composite materials possess good biological activity in vitro and superior pro-osteogenic/odontogenic potential.These materials are expected to be a new material for the repair of oral and maxillofacial bone defects.