摘要
目的基于3D打印三周期极小曲面(triply periodic minimal surfaces,TPMS)基元支架,探索不同孔径对成骨细胞MC3T3-E1增殖和分化的影响,为3D打印个性化假体多孔结构的设计提供理论基础。方法基于TPMS和选择性激光熔化(selective laser melting,SLM)设计和制造具有3种孔径(400μm、600μm和800μm)的TPMS基元支架。将不同孔径的支架与成骨细胞MC3T3-E1共培养,通过细胞计数试剂(cell counting kit-8,CCK-8)、细胞骨架染色、碱性磷酸酶(alkaline phosphatase,ALP)活性和蛋白质印迹法(western blot,WB)分析评价其对成骨细胞增殖和分化的影响。结果扫描电镜显示成功制造了不同孔径的3D打印TPMS基元支架,并且成骨细胞MC3T3-E1在支架中展现了良好的黏附性。CCK-8实验的结果表明,随着孔径的增大,MC3T3-E1的增殖性也明显增加。此外,TPMS基元支架在孔径大小为600μm时ALP的活性最高。WB分析进一步发现Ⅰ型胶原蛋白(collagen typeⅠ,ColⅠ)、Runt相关转录因子2(Runt related transcription factor 2,Runx2)和骨桥蛋白(osteopontin,OPN)在600μm组的表达同样最高。结论随着孔径的增大,3D打印TPMS基元支架更有助于成骨细胞增殖,而在孔径大小为600μm时最有助于成骨细胞分化。
Objective To explore the effects of different pore sizes of 3D printed triply periodic minimal surfaces(TPMS)primitive scaffolds on the proliferation and differentiation of osteoblasts MC3T3-E1,and to provide a theoretical basis for the design of subsequent 3D printed personalized prosthetic porous structures.Methods Design and fabrication of TPMS substrate scaffolds with three pore sizes(400μm,600μm and 800μm)were conducted based on TPMS implicit surface functions and selective laser melting(SLM).The scaffolds with different pore sizes were co-cultured with osteoblasts MC3T3-E1 and their effects on osteoblast proliferation and differentiation were evaluated by CCK-8,Cytoskeleton staining,alkaline phosphatase activity and western blot(WB)analysis.Results Scanning electron microscopy showed that TPMS primitive scaffolds with different pore sizes were successfully fabricated and osteoblasts MC3T3-E1 showed good adhesion in the scaffolds.The results of CCK-8 experiments indicated that the proliferation of MC3T3-E1 increased significantly with increasing pore size.In addition,the TPMS primitive scaffold exhibited the highest alkaline phosphatase activity at a pore size of 600μm.WB analysis further revealed that the expression of collagen typeⅠ(ColⅠ),Runt related transcription factor 2(Runx2)and osteopontin(OPN)proteins were also highest in the 600μm group.Conclusion As the pore size increased,the 3D printed TPMS scaffold demonstrates enhanced osteoblast proliferation.Furthermore,a pore size of 600μm is optimal for osteoblast differentiation.
作者
程德斌
张昭
付军
刘冬
范宏斌
Cheng Debin;Zhang Zhao;Fu Jun;Liu Dong;Fan Hongbin(Department of Orthopaedics Surgery,Xijing Hospital,Air Force Military Medical University,Xi’an 710032,China)
出处
《实用骨科杂志》
2023年第11期994-1000,共7页
Journal of Practical Orthopaedics
基金
国家自然科学基金(31971272)
陕西省国际科技合作重点项目(2023-GHZD-25)
陕西省自然科学基金(2022SF-054)。
关键词
3D打印
三周期极小曲面
成骨细胞
孔径
分化
3D printing
triply periodic minimal surfaces
osteoblasts
pore size
differentiation
