Regulation of osteoblast functions on titanium surfaces with different micro/nanotopographies and compositions
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摘要
Surface modification of medical implants is considered as an effective method to improve cellular behaviors and the integration of tissues with materials. Titanium(Ti)-based materials with four different micro/nano-structures and compositions were prepared by acid etching, electrochemical anodization and alkali-heat treatment. The surface morphologies and compositions of the different surface-modified Ti materials were characterized by field-emission scanning electron microscopy(FE-SEM),atomic force microscopy(AFM) and X-ray diffraction(XRD). The effects of the micro/nano structured and compositions of the surfaces on cellular responses were investigated in vitro by observing the morphology, adhesion, proliferation and osteogenic differentiation of osteoblasts. To further investigate the underlying mechanisms, an RT-PCR assay was performed to analyze the expression levels of cell adhesion-related genes. Our results indicated that the nanosized structure and anatase composition could promote the adhesion and proliferation of MC3T3-E1 pre-osteoblast, as well as alkaline phosphatase activity and extracellular matrix mineralization via the integrin-FAK signaling pathway. Taken together, our innovation presented in this work demonstrated that the surface nano-structure design and composition of biomedical implants can be modified of for future orthopaedic applications.
Surface modification of medical implants is considered as an effective method to improve cellular behaviors and the integration of tissues with materials. Titanium(Ti)-based materials with four different micro/nano-structures and compositions were prepared by acid etching, electrochemical anodization and alkali-heat treatment. The surface morphologies and compositions of the different surface-modified Ti materials were characterized by field-emission scanning electron microscopy(FE-SEM),atomic force microscopy(AFM) and X-ray diffraction(XRD). The effects of the micro/nano structured and compositions of the surfaces on cellular responses were investigated in vitro by observing the morphology, adhesion, proliferation and osteogenic differentiation of osteoblasts. To further investigate the underlying mechanisms, an RT-PCR assay was performed to analyze the expression levels of cell adhesion-related genes. Our results indicated that the nanosized structure and anatase composition could promote the adhesion and proliferation of MC3T3-E1 pre-osteoblast, as well as alkaline phosphatase activity and extracellular matrix mineralization via the integrin-FAK signaling pathway. Taken together, our innovation presented in this work demonstrated that the surface nano-structure design and composition of biomedical implants can be modified of for future orthopaedic applications.
Surface modification of medical implants is considered as an effective method to improve cellular behaviors and the integration of tissues with materials. Titanium(Ti)-based materials with four different micro/nano-structures and compositions were prepared by acid etching, electrochemical anodization and alkali-heat treatment. The surface morphologies and compositions of the different surface-modified Ti materials were characterized by field-emission scanning electron microscopy(FE-SEM),atomic force microscopy(AFM) and X-ray diffraction(XRD). The effects of the micro/nano structured and compositions of the surfaces on cellular responses were investigated in vitro by observing the morphology, adhesion, proliferation and osteogenic differentiation of osteoblasts. To further investigate the underlying mechanisms, an RT-PCR assay was performed to analyze the expression levels of cell adhesion-related genes. Our results indicated that the nanosized structure and anatase composition could promote the adhesion and proliferation of MC3T3-E1 pre-osteoblast, as well as alkaline phosphatase activity and extracellular matrix mineralization via the integrin-FAK signaling pathway. Taken together, our innovation presented in this work demonstrated that the surface nano-structure design and composition of biomedical implants can be modified of for future orthopaedic applications.
引文
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2 Neoh K G,Hu X,Zheng D,et al.Balancing osteoblast functions and bacterial adhesion on functionalized titanium surfaces.Biomaterials,2012,33:2813-2822
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5 Ning C,Yu P,Zhu Y,et al.Built-in microscale electrostatic fields induced by anatase-rutile-phase transition in selective areas promote osteogenesis.NPG Asia Mater,2016,8:e243
6 Chen L,Li J A,Chang J W,et al.Mg-Zn-Y-Nd coated with citric acid and dopamine by layer-by-layer self-assembly to improve surface biocompatibility.Sci China Technol Sci,2018,61:1228-1237
7 Brammer K S,Oh S,Frandsen C J,et al.Nanotube surface triggers increased chondrocyte extracellular matrix production.Mater Sci EngC,2010,30:518-525
8 Bsat S,Yavari S A,Munsch M,et al.Effect of alkali-acid-heat chemical surface treatment on electron beam melted porous titanium and its apatite forming ability.Materials,2015,8:1612-1625
9 Tan G,Tan Y,Ni G,et al.Controlled oxidative nanopatterning of microrough titanium surfaces for improving osteogenic activity.JMater Sci-Mater Med,2014,25:1875-1884
10 Wang X,Gittens R A,Song R,et al.Effects of structural properties of electrospun TiO2nanofiber meshes on their osteogenic potential.Acta Biomater,2012,8:878-885
11 Huang S,Peng W,Ning C,et al.Nanostructure transition on anodic titanium:structure control via a competition strategy between electrochemical oxidation and chemical etching.J Phys Chem C,2012,116:22359-22364
12 Li M,Ma Z,Zhu Y,et al.Toward a molecular understanding of the antibacterial mechanism of copper-bearing titanium alloys against staphylococcus aureus.Adv Healthcare Mater,2016,5:557-566
13 Hao J,Li Y,Li B,et al.Biological and mechanical effects of micronanostructured titanium surface on an osteoblastic cell line in vitro and osteointegration in vivo.Appl Biochem Biotechnol,2017,183:280-292
14 Cantini M,Gomide K,Moulisova V,et al.Vitronectin as a micromanager of cell response in material-driven fibronectin nanonetworks.Adv Biosys,2017,1:1700047
15 Roach P,Farrar D,Perry C C.Surface tailoring for controlled protein adsorption:Effect of topography at the nanometer scale and chemistry.J Am Chem Soc,2006,128:3939-3945
16 Zhou J,Han Y,Lu S.Direct role of interrod spacing in mediating cell adhesion on Sr-HA nanorod-patterned coatings.IJN,2014,9:1243-1260
17 Wang K,Bruce A,Mezan R,et al.Nanotopographical modulation of cell function through nuclear deformation.ACS Appl Mater Interfaces,2016,8:5082-5092
18 He J,Zhou W,Zhou X,et al.The anatase phase of nanotopography titania plays an important role on osteoblast cell morphology and proliferation.J Mater Sci-Mater Med,2008,19:3465-3472
19 Albuschies J,Vogel V.The role of filopodia in the recognition of nanotopographies.Sci Rep,2013,3:1658
20 Ni S,Li C,Ni S,et al.Understanding improved osteoblast behavior on select nanoporous anodic alumina.Intl J Nanomed,2014,9:3325
21 Swan E E L,Popat K C,Grimes C A,et al.Fabrication and evaluation of nanoporous alumina membranes for osteoblast culture.J Biomed Mater Res,2005,72A:288-295
22 Xin T,Greco V,Myung P.Hardwiring stem cell communication through tissue structure.Cell,2016,164:1212-1225
23 Oh S,Brammer K S,Li Y S J,et al.Stem cell fate dictated solely by altered nanotube dimension.Proc Natl Acad Sci USA,2009,106:2130-2135
24 Han Y,Zhou J,Lu S,et al.Enhanced osteoblast functions of narrow interligand spaced Sr-HA nano-fibers/rods grown on microporous titania coatings.RSC Adv,2013,3:11169-11184
25 Strohmeyer N,Bharadwaj M,Costell M,et al.Fibronectin-boundα5β1 integrins sense load and signal to reinforce adhesion in less than a second.Nat Mater,2017,16:1262-1270
26 Qian M,Lu Z,Chen C,et al.Alkaline nanoparticle coatings improve resin bonding of 10-methacryloyloxydecyldihydrogenphosphate-conditioned zirconia.IJN,2016,11:5057-5066
27 Zhou Q,Casta?eda Ocampo O,Guimar?es C F,et al.Screening platform for cell contact guidance based on inorganic biomaterial micro/nanotopographical gradients.ACS Appl Mater Interfaces,2017,9:31433-31445
28 Chien F C,Dai Y H,Kuo C W,et al.Flexible nanopillars to regulate cell adhesion and movement.Nanotechnology,2016,27:475101
29 Dumbauld D W,Shin H,Gallant N D,et al.Contractility modulates cell adhesion strengthening through focal adhesion kinase and assembly of vinculin-containing focal adhesions.J Cell Physiol,2010,275
30 Omachi T,Ichikawa T,Kimura Y,et al.Vinculin association with actin cytoskeleton is necessary for stiffness-dependent regulation of vinculin behavior.PLoS ONE,2017,12:e0175324
31 Matsuzawa M,Arai C,Nomura Y,et al.Periostin of human periodontal ligament fibroblasts promotes migration of human mesenchymal stem cell through theαvβ3 integrin/FAK/PI3K/Akt pathway.J Periodont Res,2015,50:855-863
32 Visavadiya N P,Keasey M P,Razskazovskiy V,et al.Integrin-FAKsignaling rapidly and potently promotes mitochondrial function through STAT3.Cell Commun Signal,2016,14:32
33 Lipski A M,Pino C J,Haselton F R,et al.The effect of silica nanoparticle-modified surfaces on cell morphology,cytoskeletal organization and function.Biomaterials,2008,29:3836-3846
34 Sandbo N,Smolyaninova L V,Orlov S N,et al.Control of myofibroblast differentiation and function by cytoskeletal signaling.Biochem Moscow,2016,81:1698-1708
35 Li M,Yang X,Wang W,et al.Evaluation of the osteo-inductive potential of hollow three-dimensional magnesium-strontium substitutes for the bone grafting application.Mater Sci Eng-C,2016,73:347-356
36 Cha K J,Hong J M,Cho D W,et al.Enhanced osteogenic fate and function of MC3T3-E1 cells on nanoengineered polystyrene surfaces with nanopillar and nanopore arrays.Biofabrication,2013,5:025007
37 Wei Y,Mo X,Zhang P,et al.Directing stem cell differentiation via electrochemical reversible switching between nanotubes and nanotips of polypyrrole array.ACS Nano,2017,11:5915-5924
38 Xu J,Chen H,Li X,et al.Optimal intensity shock wave promotes the adhesion and migration of rat osteoblasts via integrinβ1-mediated expression of phosphorylated focal adhesion kinase.J Biol Chem,2012,287:26200-26212
39 Gongadze E,Kabaso D,Bauer S,et al.Adhesion of osteoblasts to a nanorough titanium implant surface.Intl J Nanomed,2011,6:1801-1816
40 Siebers M C,ter Brugge P J,Walboomers X F,et al.Integrins as linker proteins between osteoblasts and bone replacing materials:A critical review.Biomaterials,2005,26:137-146
41 Rosa A L,Kato R B,Castro Raucci L M S,et al.Nanotopography drives stem cell fate toward osteoblast differentiation throughα1-β1integrin signaling pathway.J Cell Biochem,2014,115:540-548
42 Wei M,Li S,Le W.Nanomaterials modulate stem cell differentiation:Biological interaction and underlying mechanisms.J Nanobiotechnol,2017,15:75
43 Smith L A,Liu X,Hu J,et al.The enhancement of human embryonic stem cell osteogenic differentiation with nano-fibrous scaffolding.Biomaterials,2010,31:5526-5535
44 Simann M,Le Blanc S,Schneider V,et al.Canonical FGFs prevent osteogenic lineage commitment and differentiation of human bone marrow stromal cells via erk1/2 signaling.J Cell Biochem,2017,118:263-275
45 Mahmood M,Li Z,Casciano D,et al.Nanostructural materials increase mineralization in bone cells and affect gene expression through miRNA regulation.J Cellular Mol Med,2011,15:2297-2306
2 Neoh K G,Hu X,Zheng D,et al.Balancing osteoblast functions and bacterial adhesion on functionalized titanium surfaces.Biomaterials,2012,33:2813-2822
3 Sundfeldt M,Carlsson L V,Johansson C B,et al.Aseptic loosening,not only a question of wear:A review of different theories.Acta Orthopaedica,2006,77:177-197
4 Boos C,Fink K,Stomberg P,et al.Der Einfluss der Knochenmineraldichte und des Verankerungsverfahrens auf die Prim?rstabilit?t von zementfrei implantierten Tibiakomponenten/The influence of bone quality and the fixation procedure on the primary stability of cementless implanted tibial plateaus.Biomedizinische Technik/BioMed Eng,2008,53:70-76
5 Ning C,Yu P,Zhu Y,et al.Built-in microscale electrostatic fields induced by anatase-rutile-phase transition in selective areas promote osteogenesis.NPG Asia Mater,2016,8:e243
6 Chen L,Li J A,Chang J W,et al.Mg-Zn-Y-Nd coated with citric acid and dopamine by layer-by-layer self-assembly to improve surface biocompatibility.Sci China Technol Sci,2018,61:1228-1237
7 Brammer K S,Oh S,Frandsen C J,et al.Nanotube surface triggers increased chondrocyte extracellular matrix production.Mater Sci EngC,2010,30:518-525
8 Bsat S,Yavari S A,Munsch M,et al.Effect of alkali-acid-heat chemical surface treatment on electron beam melted porous titanium and its apatite forming ability.Materials,2015,8:1612-1625
9 Tan G,Tan Y,Ni G,et al.Controlled oxidative nanopatterning of microrough titanium surfaces for improving osteogenic activity.JMater Sci-Mater Med,2014,25:1875-1884
10 Wang X,Gittens R A,Song R,et al.Effects of structural properties of electrospun TiO2nanofiber meshes on their osteogenic potential.Acta Biomater,2012,8:878-885
11 Huang S,Peng W,Ning C,et al.Nanostructure transition on anodic titanium:structure control via a competition strategy between electrochemical oxidation and chemical etching.J Phys Chem C,2012,116:22359-22364
12 Li M,Ma Z,Zhu Y,et al.Toward a molecular understanding of the antibacterial mechanism of copper-bearing titanium alloys against staphylococcus aureus.Adv Healthcare Mater,2016,5:557-566
13 Hao J,Li Y,Li B,et al.Biological and mechanical effects of micronanostructured titanium surface on an osteoblastic cell line in vitro and osteointegration in vivo.Appl Biochem Biotechnol,2017,183:280-292
14 Cantini M,Gomide K,Moulisova V,et al.Vitronectin as a micromanager of cell response in material-driven fibronectin nanonetworks.Adv Biosys,2017,1:1700047
15 Roach P,Farrar D,Perry C C.Surface tailoring for controlled protein adsorption:Effect of topography at the nanometer scale and chemistry.J Am Chem Soc,2006,128:3939-3945
16 Zhou J,Han Y,Lu S.Direct role of interrod spacing in mediating cell adhesion on Sr-HA nanorod-patterned coatings.IJN,2014,9:1243-1260
17 Wang K,Bruce A,Mezan R,et al.Nanotopographical modulation of cell function through nuclear deformation.ACS Appl Mater Interfaces,2016,8:5082-5092
18 He J,Zhou W,Zhou X,et al.The anatase phase of nanotopography titania plays an important role on osteoblast cell morphology and proliferation.J Mater Sci-Mater Med,2008,19:3465-3472
19 Albuschies J,Vogel V.The role of filopodia in the recognition of nanotopographies.Sci Rep,2013,3:1658
20 Ni S,Li C,Ni S,et al.Understanding improved osteoblast behavior on select nanoporous anodic alumina.Intl J Nanomed,2014,9:3325
21 Swan E E L,Popat K C,Grimes C A,et al.Fabrication and evaluation of nanoporous alumina membranes for osteoblast culture.J Biomed Mater Res,2005,72A:288-295
22 Xin T,Greco V,Myung P.Hardwiring stem cell communication through tissue structure.Cell,2016,164:1212-1225
23 Oh S,Brammer K S,Li Y S J,et al.Stem cell fate dictated solely by altered nanotube dimension.Proc Natl Acad Sci USA,2009,106:2130-2135
24 Han Y,Zhou J,Lu S,et al.Enhanced osteoblast functions of narrow interligand spaced Sr-HA nano-fibers/rods grown on microporous titania coatings.RSC Adv,2013,3:11169-11184
25 Strohmeyer N,Bharadwaj M,Costell M,et al.Fibronectin-boundα5β1 integrins sense load and signal to reinforce adhesion in less than a second.Nat Mater,2017,16:1262-1270
26 Qian M,Lu Z,Chen C,et al.Alkaline nanoparticle coatings improve resin bonding of 10-methacryloyloxydecyldihydrogenphosphate-conditioned zirconia.IJN,2016,11:5057-5066
27 Zhou Q,Casta?eda Ocampo O,Guimar?es C F,et al.Screening platform for cell contact guidance based on inorganic biomaterial micro/nanotopographical gradients.ACS Appl Mater Interfaces,2017,9:31433-31445
28 Chien F C,Dai Y H,Kuo C W,et al.Flexible nanopillars to regulate cell adhesion and movement.Nanotechnology,2016,27:475101
29 Dumbauld D W,Shin H,Gallant N D,et al.Contractility modulates cell adhesion strengthening through focal adhesion kinase and assembly of vinculin-containing focal adhesions.J Cell Physiol,2010,275
30 Omachi T,Ichikawa T,Kimura Y,et al.Vinculin association with actin cytoskeleton is necessary for stiffness-dependent regulation of vinculin behavior.PLoS ONE,2017,12:e0175324
31 Matsuzawa M,Arai C,Nomura Y,et al.Periostin of human periodontal ligament fibroblasts promotes migration of human mesenchymal stem cell through theαvβ3 integrin/FAK/PI3K/Akt pathway.J Periodont Res,2015,50:855-863
32 Visavadiya N P,Keasey M P,Razskazovskiy V,et al.Integrin-FAKsignaling rapidly and potently promotes mitochondrial function through STAT3.Cell Commun Signal,2016,14:32
33 Lipski A M,Pino C J,Haselton F R,et al.The effect of silica nanoparticle-modified surfaces on cell morphology,cytoskeletal organization and function.Biomaterials,2008,29:3836-3846
34 Sandbo N,Smolyaninova L V,Orlov S N,et al.Control of myofibroblast differentiation and function by cytoskeletal signaling.Biochem Moscow,2016,81:1698-1708
35 Li M,Yang X,Wang W,et al.Evaluation of the osteo-inductive potential of hollow three-dimensional magnesium-strontium substitutes for the bone grafting application.Mater Sci Eng-C,2016,73:347-356
36 Cha K J,Hong J M,Cho D W,et al.Enhanced osteogenic fate and function of MC3T3-E1 cells on nanoengineered polystyrene surfaces with nanopillar and nanopore arrays.Biofabrication,2013,5:025007
37 Wei Y,Mo X,Zhang P,et al.Directing stem cell differentiation via electrochemical reversible switching between nanotubes and nanotips of polypyrrole array.ACS Nano,2017,11:5915-5924
38 Xu J,Chen H,Li X,et al.Optimal intensity shock wave promotes the adhesion and migration of rat osteoblasts via integrinβ1-mediated expression of phosphorylated focal adhesion kinase.J Biol Chem,2012,287:26200-26212
39 Gongadze E,Kabaso D,Bauer S,et al.Adhesion of osteoblasts to a nanorough titanium implant surface.Intl J Nanomed,2011,6:1801-1816
40 Siebers M C,ter Brugge P J,Walboomers X F,et al.Integrins as linker proteins between osteoblasts and bone replacing materials:A critical review.Biomaterials,2005,26:137-146
41 Rosa A L,Kato R B,Castro Raucci L M S,et al.Nanotopography drives stem cell fate toward osteoblast differentiation throughα1-β1integrin signaling pathway.J Cell Biochem,2014,115:540-548
42 Wei M,Li S,Le W.Nanomaterials modulate stem cell differentiation:Biological interaction and underlying mechanisms.J Nanobiotechnol,2017,15:75
43 Smith L A,Liu X,Hu J,et al.The enhancement of human embryonic stem cell osteogenic differentiation with nano-fibrous scaffolding.Biomaterials,2010,31:5526-5535
44 Simann M,Le Blanc S,Schneider V,et al.Canonical FGFs prevent osteogenic lineage commitment and differentiation of human bone marrow stromal cells via erk1/2 signaling.J Cell Biochem,2017,118:263-275
45 Mahmood M,Li Z,Casciano D,et al.Nanostructural materials increase mineralization in bone cells and affect gene expression through miRNA regulation.J Cellular Mol Med,2011,15:2297-2306