不同孔径3D打印三周期极小曲面基元支架调控成骨细胞生物学行为的研究

摘要
图/表
参考文献
相关文章 (15)

全文: PDF (7071 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS)

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(400 μm、600 μm和800 μm)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600 μm时ALP的活性最高。WB分析进一步发现Ⅰ型胶原蛋白(collagen type Ⅰ，Col Ⅰ)、Runt相关转录因子2(Runt related transcription factor 2，Runx2)和骨桥蛋白(osteopontin，OPN)在600 μm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600 μm时最有助于成骨细胞分化。

	服务

	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	程德斌
	张昭
	付军
	刘冬
	范宏斌

关键词 ： 3D打印, 三周期极小曲面, 成骨细胞, 孔径, 分化

Abstract：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 (400 μm，600 μm and 800 μm) 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 600 μm.WB analysis further revealed that the expression of collagen type Ⅰ(Col Ⅰ)，Runt related transcription factor 2(Runx2) and osteopontin (OPN) proteins were also highest in the 600 μm group. Conclusion As the pore size increased，the 3D printed TPMS scaffold demonstrates enhanced osteoblast proliferation.Furthermore，a pore size of 600 μm is optimal for osteoblast differentiation.

Key words： 3D printing triply periodic minimal surfaces osteoblasts pore size differentiation

基金资助: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)

通讯作者: 范宏斌

作者简介: 程德斌(1997－ )，男，研究生在读，空军军医大学附属西京医院骨科，chengdb11@163.com

引用本文:

程德斌，张昭，付军，刘冬，范宏斌. 不同孔径3D打印三周期极小曲面基元支架调控成骨细胞生物学行为的研究[J]. 实用骨科杂志, 2023, 29(11): 994-.
Cheng Debin，Zhang Zhao，Fu Jun，Liu Dong，Fan Hongbin. Modulation of Osteoblast Biological Behavior by 3D Printed Triply Periodic Minimal Surfaces Primitive Scaffolds with Different Pore Sizes. sygkzz, 2023, 29(11): 994-.

链接本文:

http://www.sygkzz.com/CN/ 或 http://www.sygkzz.com/CN/Y2023/V29/I11/994

［1］张正也，刘晓奇，庄金鹏，等.3D打印钛合金在骨科的临床应用及进展［J］.实用骨科杂志，2021，27(9)：820-824.
［2］Sikavitsas VI，Temenoff JS，Mikos AG.Biomaterials and bone mechanotransduction［J］.Biomaterials，2001，22(19)：2581-2593.
［3］Yan C，Hao L，Hussein A，et al.Ti6Al-4V triply periodic minimal surface structures for bone implants fabricated via selective laser melting［J］.J Mech Behav Biomed Mater，2015(51)：61-73.
［4］Zhang T，Wei Q，Zhou H，et al.Threedimensional-printed individualized porous implants：A new "implantbone" interface fusion concept for large bone defect treatment［J］.Bioact Mater，2021，6(11)：3659-3670.
［5］Yao Y，Yang Y，Ye Q，et al.Effects of pore size and porosity on cytocompatibility and osteogenic differentiation of porous titanium［J］.J Mater Sci Mater Med，2021，32(6)：72.
［6］Taniguchi N，Fujibayashi S，Takemoto M，et al.Effect of pore size on bone ingrowth into porous titanium implants fabricated by additive manufacturing：An in vivo experiment［J］.Mater Sci Eng C Mater Biol Appl，2016(59)：690-701.
［7］Frosch K，Barvencik F，Lohmann CH，et al.Migration，matrix production and lamellar bone formation of human osteoblast-like cells in porous titanium implants［J］.Cells Tissues Organs，2002，170(4)：214-227.
［8］Kapfer SC，Hyde ST，Mecke K，et al.Minimal surface scaffold designs for tissue engineering.［J］.Biomaterials，2011，32(29)：6875-6882.
［9］Li L，Shi J，Zhang K，et al.Early osteointegration evaluation of porous Ti6Al4V scaffolds designed based on triply periodic minimal surface models［J］.J Orthop Translat，2019(19)：94-105.
［10］Jinnai H，Watashiba H，Kajihara T，et al.Surface curvatures of trabecular bone microarchitecture［J］.Bone，2002，30(1)：191-194.
［11］刘佳辛，贾鹏，门玉涛，等.基于三周期极小曲面骨小梁结构的设计及优化［J］.中国组织工程研究，2023，27(7)：992-997.
［12］Zekry KM，Yamamoto N，Hayashi K，et al.Reconstruction of intercalary bone defect after resection of malignant bone［J］.J Orthop Surg (HK)，2019，27(1)：615493258.
［13］Chen Z，Yan X，Yin S，et al.Influence of the pore size and porosity of selective laser melted Ti6Al4V ELI porous scaffold on cell proliferation，osteogenesis and bone ingrowth［J］.Mater Sci Eng C Mater Biol Appl，2020(106)：110289.
［14］Wang C，Xu D，Li S，et al.Effect of pore size on the physicochemical properties and osteogenesis of Ti6Al4V porous scaffolds with bionic structure［J］.ACS Omega，2020，5(44)：28684-28692.
［15］Karimipour-Fard P，Behravesh AH，JonesTaggart H，et al.Effects of design，porosity and biodegradation on mechanical and morphological properties of additivemanufactured triply periodic minimal surface scaffolds［J］.J Mech Behav Biomed Mater，2020(112)：104064.
［16］Wang N，Meenashisundaram GK，Kandilya D，et al.A biomechanical evaluation on cubic，octet，and TPMS gyroid Ti6Al4V lattice structures fabricated by selective laser melting and the effects of their debris on human osteoblast-like cells［J］.Biomaterials advances，2022(137)：212829.
［17］Castro APG，Ruben RB，Goncalves SB，et al.Numerical and experimental evaluation of TPMS gyroid scaffolds for bone tissue［J］.Comput Methods Biomech Biomed Engin，2019，22(6)：567-573.
［18］张昭，朱东泽，党竞医，等.3D打印技术在骨肿瘤精准化治疗中的研究进展［J］.生物骨科材料与临床研究，2022，19(3)：81-85.
［19］张建国，赵晟，胡凤玲，等.SLM打印Ti6Al4V不同晶格结构多孔材料的力学性能［J］.有色金属工程，2023，13(1)：10-17.
［20］李海亮，贾德昌，杨治华，等.选区激光熔化3D打印钛合金及其复合材料研究进展［J］.材料科学与工艺，2019，27(2)：1-15.
［21］Szatkiewicz T，Laskowska D，Ba-asz B，et al.The influence of the structure parameters on the mechanical properties of cylindrically mapped gyroid TPMS fabricated by selective laser melting with 316L stainless steel powder［J］.Materials (Basel，Switzerland)，2022，15(12)：4352.
［22］Li Z，Chen Z，Chen X，et al.Effect of unit configurations and parameters on the properties of Ti-6Al-4V unit-stacked scaffolds：A trade-off between mechanical and permeable performance［J］.J Mech Behav Biomed Mater，2021(116)：104332.
［23］Li X，Chen T，Hu J，et al.Modified surface morphology of a novel Ti-24Nb-4Zr-7.9Sn titanium alloy via anodic oxidation for enhanced interfacial biocompatibility and osseointegration［J］.Colloids Surf B Biointerfaces，2016(144)：265-275.
［24］Sollazzo V，Pezzetti F，Massari L，et al.Evaluation of gene expression in MG63 human osteoblastlike cells exposed to tantalum powder by microarray technology［J］.Int J Periodontics Restorative Dent，2011，31(4)：e17-e28.
［25］Kapat K，Srivas PK，Rameshbabu AP，et al.Influence of porosity and pore-size distribution in Ti6Al4 V foam on physicomechanical properties，osteogenesis，and quantitative validation of bone ingrowth by microcomputed tomography［J］.ACS Appl Mater Interfaces，2017，9(45)：39235-39248.
［26］Wang C，Xu D，Lin L，et al.Large-pore-size Ti6Al4V scaffolds with different pore structures for vascularized bone regeneration［J］.Mater Sci Eng C Mater Biol Appl，2021(131)：112499.
［27］Bueno Rde B，Adachi P，Castro-Raucci LM，et al.Oxidative nanopatterning of titanium surfaces promotes production and extracellular accumulation of osteopontin［J］.Braz Dent J，2011，22(3)：179-184.
［28］Vimalraj S.Alkaline phosphatase：Structure，expression and its function in bone mineralization［J］.Gene，2020(754)：144855.
［29］Zhu Y，Zheng T，Wen L，et al.Osteogenic capability of strontium and icariin-loaded TiO(2) nanotube coatings in vitro and in osteoporotic rats［J］.J Biomater Appl，2021，35(9)：1119-1131.
［30］宋芹，颜军，郭晓强，等.补骨脂与罗非鱼鳞胶原蛋白多肽对MC3T3-E1细胞ALP、Ⅰ型胶原蛋白mRNA表达的影响［J］.中药药理与临床，2011，27(1)：47-49.
［31］Xu J，Li Z，Hou Y，et al.Potential mechanisms underlying the Runx2 induced osteogenesis of bone marrow［J］.Am J Transl Res，2015，7(12)：2527-2535.
［32］Dalle Carbonare L，Innamorati G，Valenti MT.Transcription factor Runx2 and its application to bone tissue engineering［J］.Stem Cell Rev Rep，2012，8(3)：891-897.
［33］Si J，Wang C，Zhang D，et al.Osteopontin in bone metabolism and bone diseases［J］.Med Sci Monit，2020(26)：e919159.
［34］Zhao H，Shen S，Zhao L，et al.3D printing of dual-cell delivery titanium alloy scaffolds for improving osseointegration through enhancing angiogenesis and osteogenesis［J］.BMC Musculoskelet Disord，2021，22(1)：734.
［35］Ran Q，Yang W，Hu Y，et al.Osteogenesis of 3D printed porous Ti6Al4V implants with different pore sizes［J］.J Mech Behav Biomed Mater，2018(84)：1-11.
［36］Myakinin A，Turlybekuly A，Pogrebnjak A，et al.In vitro evaluation of electrochemically bioactivated Ti6Al4V 3D porous scaffolds［J］.Mater Sci Eng C Mater Biol Appl，2021(121)：111870.
［37］Callens SJP，Uyttendaele RJC，Fratila-Apachitei LE，et al.Substrate curvature as a cue to guide spatiotemporal cell and tissue organization［J］.Biomaterials，2020(232)：119739.
［38］Vlad MD，Fernández Aguado E，Gómez González S，et al.Novel titanium-apatite hybrid scaffolds with spongy bone-like micro architecture intended for spinal application：In vitro and in vivo study［J］.Mater Sci Eng C Mater Biol Appl，2020(110)：110658.