In Vitro Bone Cell Behavior on Porous Titanium Samples: Influence of Porosity by Loose Sintering and Space Holder Techniques
Abstract
:1. Introduction
2. Materials and Methods
2.1. Microstructural and Macro-Mechanical Behavior
2.2. In-Vitro Cell Experiments
2.2.1. Cell Adhesion and Proliferation of Myoblast Cells
2.2.2. In-Vitro Evaluation of Osteoblast Response
2.2.3. Statistical Analysis
3. Results
3.1. Porosity and Mechanical Behavior
3.2. Cell Adhesion and Proliferation
4. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- Brunette, D.M.; Tengvall, P.; Textor, M.; Thomsen, P. Titanium in Medicine: Materials Science, Surface Science, Engineering, Biological Responses and Medical Applications; Springer: Berlin, German, 2001. [Google Scholar]
- Van-Noort, R. Titanium: The implant material of today. J. Mater. Sci. 1987, 22, 3801. [Google Scholar] [CrossRef]
- Williams, D.F. On the mechanism of biocompatibility. Biomaterials 2008, 29, 1–13. [Google Scholar] [CrossRef] [PubMed]
- He, G.; Liu, P.; Tan, Q. Porous titanium materials with entangled wire structure for load-bearing biomedical applications. J. Mech. Behav. Biomed. Mater. 2012, 5, 16–31. [Google Scholar] [CrossRef] [PubMed]
- Dizlek, M.E.; Guden, M.; Turkan, U.; Tasdemirci, A. Processing and compression testing of Ti6Al4V foams for biomedical applications. J. Mater. Sci. 2009, 44, 1512. [Google Scholar] [CrossRef] [Green Version]
- Zhao, X.; Sun, H.; Lan, L.; Huang, J.; Zhang, H.; Wang, Y. Pore structures of high-porosity NiTi alloys made from elemental powders with NaCl temporary space-holders. Mater. Lett. 2009, 63, 2402. [Google Scholar] [CrossRef]
- Ryan, G.E.; Pandit, A.S.; Apatsidis, D.P. Fabrications Methods of porous metals for use in orthopaedic applications. Biomaterials 2006, 27, 2651–2670. [Google Scholar] [CrossRef]
- Bram, M.; Stiller, C.; Buchkremer, H.P.; Stover, D.; Baur, H. High-porosity titanium, stainless steel, and superalloy parts. Adv. Eng. Mater. 2000, 2, 196–199. [Google Scholar] [CrossRef]
- Dunand, D.C. Processing of titanium foams. Adv. Eng. Mater. 2004, 6, 369–376. [Google Scholar] [CrossRef]
- Wen, C.E.; Yamada, Y.; Shimojima, K.; Chino, Y.; Asahina, T.; Mabuchi, M. Processing and Mechanical Properties of Autogenous Titanium Implant Materials. Mater. Sci. Mater. Med. 2002, 13, 397–401. [Google Scholar] [CrossRef]
- Singh, R.; Lee, P.D.; Dashwood, R.J.; Lindley, T.C. Titanium foams for biomedical applications: A review. Mater. Technol. 2010, 25, 127–136. [Google Scholar] [CrossRef]
- Niinomi, M.; Nakai, M.; Hieda, J. Development of new metallic alloys for biomedical applications. Acta Biomater. 2012, 8, 3888–3903. [Google Scholar] [CrossRef] [PubMed]
- Gepreel, M.A.-H.; Niinomi, M. Biocompatibility of Ti-alloys for long-term implantation. J. Mech. Behav. Biomed. Mater. 2013, 20, 407–415. [Google Scholar] [CrossRef] [PubMed]
- Stamboulis, A.G.; Boccaccini, A.R.; Hench, L.L. Novel biodegradable polymer/bioactive glass composites for tissue engineering applications. Adv. Eng. Mater. 2002, 4, 105–109. [Google Scholar] [CrossRef]
- Domínguez-Trujillo, C.; Ternero, F.; Rodríguez-Ortiz, J.A.; Heise, S.; Boccaccini, A.R.; Lebrato, J.; Torres, Y. Bioactive coatings on porous titanium for biomedical applications. Surf. Coat. Technol. 2018, 349, 584–592. [Google Scholar] [CrossRef]
- Hsu, H.-C.; Hsu, S.-K.; Wu, S.-C.; Wang, P.-H.; Ho, W.-F. Design and characterization of highly porous titanium foams with bioactive surface sintering in air. J. Alloys Compd. 2013, 575, 326–332. [Google Scholar] [CrossRef]
- Ye, B.; Dunand, D.C. Titanium foams produced by solid-state replication of NaCl powders. Mater. Sci. Eng. A 2010, 528, 691–697. [Google Scholar] [CrossRef]
- Li, J.; Jansen, J.A.; Walboomers, X.F.; van den Beucken, J.J. Mechanical aspects of dental implants and osseointegration: A narrative review. J. Mech. Behav. Biomed. Mater. 2019, 103, 103574. [Google Scholar] [CrossRef]
- Chen, Y.; Frith, J.E.; Dehghan-Manshadi, A.; Kent, D.; Bermingham, M.; Dargusch, M. Biocompatible porous titanium scaffolds produced using a novel space holder technique. J. Biomed. Mater. Res. Part B 2018, 106, 2796–2806. [Google Scholar] [CrossRef]
- Civantos, A.; Domínguez, C.; Pino, R.J.; Setti, G.; Pavon, J.J.; Martínez-Campos, E.; Garcia, F.J.G.; Rodriguez, J.A.; Allain, J.P.; Torres, Y. Designing bioactive porous titanium interfaces to balance mechanical properties and in vitro cells behavior towards increased osseointegration. Surf. Coat. Technol. 2019, 368, 162–174. [Google Scholar] [CrossRef]
- Schmidutz, F.; Agarwal, Y.; Müller, P.; Gueorguiev, B.; Richards, R.; Sprecher, C. Stress-shielding induced bone remodeling in cementless shoulder resurfacing arthroplasty: A finite element analysis and in vivo results. J. Biomech. 2014, 47, 3509–3516. [Google Scholar] [CrossRef]
- Dabrowski, B.; Swieszkowski, W.; Godlinski, D.; Kurzydlowski, K.J. Highly porous titanium scaffolds for orthopaedic applications. J. Mech. Behav. Biomed. Mater. B: Appl. Biomater. 2010, 95, 53–61. [Google Scholar] [CrossRef] [PubMed]
- Herrera, A.; Yánez, A.; Martel, O.; Afonso, H.; Monopoli, D. Computational study and experimental validation of porous structures fabricated by electron beam melting: A challenge to avoid stress shielding. Mater. Sci. Eng. C 2014, 45, 89–93. [Google Scholar] [CrossRef]
- Muñoz, S.; Castillo, S.; Torres, Y. Different models for simulation of mechanical behaviour of porous materials. J. Mech. Behav. Biomed. Mater. 2018, 80, 88–96. [Google Scholar] [CrossRef] [PubMed]
- Arabnejad, S.; Johnston, B.; Tanzer, M.; Pasini, D. Fully porous 3D printed titanium femoral stem to reduce stress-shielding following total hip arthroplasty. J. Orthop. Res. 2017, 35, 1774–1783. [Google Scholar] [CrossRef] [PubMed]
- Lascano, S.; Arévalo, C.; Montealegre-Melendez, I.; Muñoz, S.; Rodriguez-Ortiz, J.A.; Trueba, P.; Torres, Y. Porous Titanium for Biomedical Applications: Evaluation of the Conventional Powder Metallurgy Frontier and Space-Holder Technique. Appl. Sci. 2019, 9, 982. [Google Scholar] [CrossRef] [Green Version]
- Pavón, J.J.; Trueba, P.; Rodríguez-Ortiz, J.A.; Torres, Y. Development of new titanium implants with longitudinal gradient porosity by space-holder technique. J. Mater. Sci. 2015, 50, 6103–6112. [Google Scholar] [CrossRef]
- Miao, X.; Sun, D. Graded/Gradient Porous Biomaterials. Materials 2010, 3, 26–47. [Google Scholar] [CrossRef] [Green Version]
- Torres, Y.; Lascano, S.; Bris, J.; Pavón, J.; Rodriguez, J.A. Development of porous titanium for biomedical applications: A comparison between loose sintering and space-holder techniques. Mater. Sci. Eng. C 2014, 37, 148–155. [Google Scholar] [CrossRef]
- Subramani, K.; Mathew, R.T.; Pachauri, P. Titanium surface modification techniques for dental implants—From microscale to nanoscale. In Emerging Nanotechnologies in Dentistry; Elsevier: Amsterdam, The Netherlands, 2018; pp. 99–124. [Google Scholar]
- Domínguez-Trujillo, C.; Ternero, F.; Rodríguez-Ortiz, J.A.; Pavón, J.J.; Montealegre-Meléndez, I.; Arévalo, C.; García-Moreno, F.; Torres, Y. Improvement of the balance between a reduced stress shielding and bone ingrowth by bioactive coatings onto porous titanium substrates. Surf. Coat. Technol. 2018, 338, 32–37. [Google Scholar] [CrossRef]
- Domínguez-Trujillo, C.; Beltrán, A.M.; Garvi, M.D.; Salazar-Moya, A.; Lebrato, J.; Hickey, D.J.; Rodríguez-Ortiz, J.A.; Kamm, P.H.; Lebrato, C.; García-Moreno, F. Bacterial behavior on coated porous titanium substrates for biomedical applications. Surf. Coat. Technol. 2019, 357, 896–902. [Google Scholar] [CrossRef]
- Takemoto, M.; Fujibayashi, S.; Neo, M.; So, K.; Akiyama, N.; Matsushita, T.; Kokubo, T.; Nakamura, T. A porous bioactive titanium implant for spinal interbody fusion: An experimental study using a canine model. J. Neurosurg. Spine. 2007, 7, 435. [Google Scholar] [CrossRef] [PubMed]
- Scislowska-Czarnecka, A.; Menaszek, E.; Szaraniec, B.; Kolaczkowska, E. Ceramic modifications of porous titanium: Effects on macrophage activation. Tissue Cell 2012, 44, 391–400. [Google Scholar] [CrossRef] [PubMed]
- Borjas, S.; Gil, E.J.; Cordero, L.; Pavón, J.J.; Rodriguez-Ortiz, J.A.; Boccaccini, A.R.; Torres, Y. Electrophoretic deposition and characterization of chitosan/45S5 bioactive glass composite coatings on porous titanium for biomedical applications. Key Eng. Mater. 2015, 654, 189–194. [Google Scholar] [CrossRef]
- Tobin, E.J. Recent coating developments for combination devices in orthopedic and dental applications: A literature review. Adv. Drug Deliv. Rev. 2017, 112, 88–100. [Google Scholar] [CrossRef]
- Civantos, A.; Allain, J.P.; Pavón, J.J.; Shetty, A.; El-Atwani, O.; Walker, E.; Arias, S.L.; Gordon, E.; Rodríguez-Ortiz, J.A.; Chen, M.; et al. Directed Irradiation Synthesis as an Advanced Plasma Technology for Surface Modification to Activate Porous and “as-received” Titanium Surfaces. Metals 2019, 9, 1349. [Google Scholar] [CrossRef] [Green Version]
- Wang, M.; Tang, T. Surface treatment strategies to combat implant-related infection from the beginning. J. Orthop. Transl. 2019, 17, 42–54. [Google Scholar] [CrossRef]
- Torres, Y.; Pavón, J.; Rodríguez, J. Processing and characterization of porous titanium for implants by using NaCl as space holder. J. Mater. Process. Technol. 2012, 212, 1061–1069. [Google Scholar] [CrossRef]
- Torres, Y.; Rodríguez, J.A.; Arias, S.; Echeverry, M.; Robledo, S.; Amigó, V.; Pavón, J.J. Processing, Characterization and biological testing of porous titanium obtained by space-holder technique. J. Mater. Sci. 2012, 47, 6565–6576. [Google Scholar] [CrossRef]
- Kikuchi, M.; Takahashi, M.; Okuno, O. Elastic moduli of cast Ti–Au, Ti–Ag, and Ti–Cu alloys. Dent. Mater. 2006, 22, 641–646. [Google Scholar] [CrossRef]
- Schwarz, M.L.; Kowarsch, M.; Rose, S.; Becker, K.; Lenz, T.; Jani, L. Effect of surface roughness, porosity, and a resorbable calcium phosphate coating on osseointegration of titanium in a minipig model. J. Biomed. Mater. Res. A 2009, 89, 667–678. [Google Scholar] [CrossRef]
- Le Guéhennec, L.; Soueidan, A.; Layrolle, P.; Amouriq, Y. Surface treatments of titanium dental implants for rapid osseointegration. Dent. Mater. 2007, 23, 844–854. [Google Scholar] [CrossRef] [PubMed]
- Civantos, A.; Martinez-Campos, E.; Ramos, V.; Elvira, C.; Gallardo, A.; Abarrategi, A. Titanium coatings and surface modifications: Toward clinically useful bioactive implants. ACS Biomater. Sci. Eng. 2017, 3, 1245–1261. [Google Scholar] [CrossRef]
- Martin, J.; Dean, D.D.; Cochran, D.L.; Simpson, J.; Boyan, B.; Schwartz, Z. Proliferation, differentiation, and protein synthesis of human osteoblast-like cells (MG63) cultured on previously used titanium surfaces. Clin. Oral Implants Res. 1996, 7, 27–37. [Google Scholar] [CrossRef] [PubMed]
- Izman, S.; Abdul-Kadir, M.R.; Anwar, M.; Nazim, E.; Rosliza, R.; Shah, A.; Hassan, M. Surface Modification Techniques for Biomedical Grade of Titanium Alloys: Oxidation, Carburization and Ion Implantation Processes. In Titanium Alloys–Towards Achieving Enhanced Properties for Diversified Applications; Nurul Amin, A.K.M., Ed.; Books on Demand: Rijeka, Croatia, 2012; pp. 201–228. [Google Scholar]
- Muñoz, S.; Pavón, J.; Rodríguez-Ortiz, J.A.; Civantos, A.; Allain, J.P.; Torres, Y. On the influence of space holder in the development of porous titanium implants: Mechanical, computational and biological evaluation. Mater. Charact. 2015, 108, 68–78. [Google Scholar] [CrossRef]
- Do Prado, R.F.; de Oliveira, F.S.; Nascimento, R.D.; de Vasconcellos, L.M.R.; Carvalho, Y.R.; Cairo, C.A.A. Osteoblast response to porous titanium and biomimetic surface: In vitro analysis. Mater. Sci. Eng. C 2015, 52, 194–203. [Google Scholar] [CrossRef]
- Li, G.; Wang, L.; Pan, W.; Yang, F.; Jiang, W.; Wu, X.; Kong, X.; Dai, K.; Hao, Y. In vitro and in vivo study of additive manufactured porous Ti6Al4V scaffolds for repairing bone defects. Sci. Rep. 2016, 6, 34072. [Google Scholar] [CrossRef] [Green Version]
- Romero-Gavilan, F.; Sánchez-Pérez, A.M.; Araújo-Gomes, N.; Azkargorta, M.; Iloro, I.; Elortza, F.; Gurruchaga, M.; Goñi, I.; Suay, J. Proteomic analysis of silica hybrid sol-gel coatings: A potential tool for predicting the biocompatibility of implants in vivo. Biofouling 2017, 33, 676–689. [Google Scholar] [CrossRef]
- Vilardell, A.; Cinca, N.; Garcia-Giralt, N.; Dosta, S.; Cano, I.; Nogués, X.; Guilemany, J. Osteoblastic cell response on high-rough titanium coatings by cold spray. J. Mater. Sci. Mater. Med. 2018, 29, 19. [Google Scholar] [CrossRef] [Green Version]
- Caparros, C.; Guillem-Martí, J.; Molmeneu, M.; Punset, M.; Calero, J.; Gil, F. Mechanical properties and in vitro biological response to porous titanium alloys prepared for use in intervertebral implants. J. Mech. Behav. Biomed. Mater. 2014, 39, 79–86. [Google Scholar] [CrossRef]
- Otsuki, B.; Takemoto, M.; Fujibayashi, S.; Neo, M.; Kokubo, T.; Nakamura, T. Pore throat size and connectivity determine bone and tissue ingrowth into porous implants: Three-dimensional micro-CT based structural analyses of porous bioactive titanium implants. Biomaterials 2006, 27, 5892–5900. [Google Scholar] [CrossRef] [Green Version]
- Anderson, H.C. Molecular biology of matrix vesicles. Clin. Orthop. Relat. Res. 1995, 266–280. [Google Scholar] [CrossRef]
- Anderson, H.C.; Garimella, R.; Tague, S.E. The role of matrix vesicles in growth plate development and biomineralization. Front. Biosci. 2005, 10, 822–837. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Torres-Sanchez, C.; Al Mushref, F.; Norrito, M.; Yendall, K.; Liu, Y.; Conway, P.P. The effect of pore size and porosity on mechanical properties and biological response of porous titanium scaffolds. Mater. Sci. Eng. C 2017, 77, 219–228. [Google Scholar] [CrossRef] [PubMed] [Green Version]
Route | Size Distribution, d[10], d[50], d[90] (μm) | Compaction Pressure (MPa) | Spacer Removal Procedure | Sintering Temperature (°C) | |||
---|---|---|---|---|---|---|---|
Titanium Powder | Spacer Particles | ||||||
Loose Sintering | 9.7, 23.3, 48.4 | None | 0 | None | 1000 | ||
1100 | |||||||
Space Holder | NaCl | 30 vol.% | 183, 384, 701 | 800 | Distilled water, without stirring, 50 °C, during 16 h | 1250 | |
50 vol.% | |||||||
NH4HCO3 | 30 vol.% | 73, 233, 497 | Two steps: 60 °C and 110 °C. Both in low vacuum conditions (~10−2 mbar) and 12 h | ||||
50 vol.% | |||||||
40 vol.% | 100-200 119, 184, 286 | ||||||
355-500 355, 424, 564 |
Physical and Mechanical Properties | Loose Sintering | Space Holder | |||||||
---|---|---|---|---|---|---|---|---|---|
NaCl | NH4HCO3 | ||||||||
1000 °C | 1100 °C | 30 (vol.%) | 50 (vol.%) | 30 (vol.%) | 40 (vol. %) | 50 (vol.%) | |||
100–200 µm | 355–500 µm | ||||||||
Archimedes’ method | PT (%) | 44.1 ± 1.7 | 34.3 ± 0.5 | 28.5 ± 1.0 | 47.2 ± 0.9 | 27.8 ± 0.9 | 40.2 ± 0.6 | 40.8 ± 0.5 | 44.7 ± 0.8 |
Pi (%) | 42.1 ± 5.1 | 31.0 ± 4.8 | 20.5 ± 0.8 | 44.1 ± 0.9 | 22.4 ± 0.9 | 27.9 ± 0.8 | 32.9 ± 0.7 | 38.5 ± 0.8 | |
Density (g/cm3) | 2.5 ± 0.2 | 2.8 ± 0.2 | 3.2 ± 0.2 | 2.2 ± 0.2 | 3.6 ± 0.2 | 2.69 ± 0.02 | 2.67 ± 0.02 | 2.50 ± 0.2 | |
Image analysis | PIA (%) | 43.0 ± 1.2 | 36.9 ± 4.0 | 28.4 ± 4.6 | 47.9 ± 3.1 | 29.1 ± 3.8 | 41.6 ± 3.3 | 43.8 ± 2.4 | 44.0 ± 4.0 |
Deq (µm) | 16 ± 17 | 19 ± 16 | 370 ± 244 | 392 ± 260 | 230 ± 202 | 226 ± 178 | 295 ± 287 | 245 ± 223 | |
Ff | 0.76 ± 0.11 | 0.85 ± 0.09 | 0.81 ± 0.13 | 0.70 ± 0.14 | 0.83 ± 0.11 | 0.71 ± 0.12 | 0.67 ± 0.06 | 0.79 ± 0.08 | |
Uniaxial Compression | Ec (GPa) | 13.4 ± 6 | 21.0 ± 4 | 6.6 ± 5 | 5.2 ± 5.0 | 15.9 ± 4.1 | - | 8.5 ± 7.0 | |
σy (MPa) | 128 ± 6 | 206 ± 5 | 323 ± 10 | 117 ± 12 | 298 ± 8 | 235 ± 6 | 190 ± 5 | 149 ± 9 | |
Ultrasound | Ed (GPa) | 42.2 ± 1.0 | 52.1 ± 1.2 | 58.9 ± 0.9 | 39.5 ± 1.3 | 59.8 ± 1.2 | 45.9 ± 0.7 | 45.3± 0.8 | 41.7 ± 1.0 |
© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Civantos, A.; Giner, M.; Trueba, P.; Lascano, S.; Montoya-García, M.-J.; Arévalo, C.; Vázquez, M.Á.; Allain, J.P.; Torres, Y. In Vitro Bone Cell Behavior on Porous Titanium Samples: Influence of Porosity by Loose Sintering and Space Holder Techniques. Metals 2020, 10, 696. https://doi.org/10.3390/met10050696
Civantos A, Giner M, Trueba P, Lascano S, Montoya-García M-J, Arévalo C, Vázquez MÁ, Allain JP, Torres Y. In Vitro Bone Cell Behavior on Porous Titanium Samples: Influence of Porosity by Loose Sintering and Space Holder Techniques. Metals. 2020; 10(5):696. https://doi.org/10.3390/met10050696
Chicago/Turabian StyleCivantos, Ana, Mercè Giner, Paloma Trueba, Sheila Lascano, María-José Montoya-García, Cristina Arévalo, María Ángeles Vázquez, Jean Paul Allain, and Yadir Torres. 2020. "In Vitro Bone Cell Behavior on Porous Titanium Samples: Influence of Porosity by Loose Sintering and Space Holder Techniques" Metals 10, no. 5: 696. https://doi.org/10.3390/met10050696