Different Methods to Modify the Hydrophilicity of Titanium Implants with Biomimetic Surface Topography to Induce Variable Responses in Bone Marrow Stromal Cells
"> Figure 1
<p>UV treatment effect on surface properties. Implant surface characterization shows increased wettability following treatment with UV–plasma-based cleaner. Sessile water droplet test of Ti6Al4V surface (<b>A</b>) and Ti6Al4V surface treated with UV–plasma cleaner (<b>B</b>). Contact angle measurements of water droplets for treated and untreated surfaces (<b>C</b>); measures were taken at 6 different locations on the implant surface. Optical profilometry measurements of surface micro-roughness (<b>D</b>) and peak-to-valley height (<b>E</b>). X-ray photoelectron spectroscopy to assess concentrations of elements present on the surface (<b>F</b>). Results are the means of 6 measurements taken at different points on 2 surfaces (<span class="html-italic">n</span> = 12) with bars showing SEM. Groups labeled with “*” are statistically significant compared to untreated Ti6Al4V using a Student’s unpaired <span class="html-italic">t</span>-test. (* = α < 0.05, *** = α < 0.0005, **** = α < 0.0001).</p> "> Figure 2
<p>UV treatment effect on cell response. In vitro assessment of bMSCs cultured on UV–plasma-treated and untreated Ti6Al4V surfaces. Total DNA content measured at 7 days of culture (<b>A</b>). ELISA quantification of osteoblast maturation markers osteocalcin (<b>B</b>) and osteopontin (<b>C</b>), and paracrine signaling factors osteoprotegerin (<b>D</b>) and vascular endothelial growth factor (<b>E</b>) in response to UV–plasma-treated surfaces. Immunomodulatory cytokine production of IL-6 (<b>F</b>) and IL-10 (<b>G</b>). Groups are means of 6 cultures/variables, with errors bars representing SEM. Factor production in the conditioned media was normalized to total DNA and statistics were determined by ANOVA with Tukey post-test. Groups labeled with “*” are statistically significant compared to TCPS at <span class="html-italic">p</span>-value equal to or less than 0.05.</p> "> Figure 3
<p>DBD treatment effect on surface properties. Implant surface characterization shows increased wettability following treatment with argon-based plasma cleaning method. Sessile water droplet test of Ti6Al4V surface (<b>A</b>) and Ti6Al4V surface treated with argon plasma cleaner (<b>B</b>). Contact angle measurements of water droplets for treated and untreated surfaces (<b>C</b>); measures were taken at 6 different locations on the implant surface. Optical profilometry measurements of surface micro-roughness (<b>D</b>) and peak-to-valley height (<b>E</b>). X-ray photoelectron spectroscopy to assess concentrations of elements present on the surface (<b>F</b>). Results are the means of 6 measurements taken at different points on 2 surfaces (<span class="html-italic">n</span> = 12) with bars showing SEM. Groups labeled with “*” are statistically significant compared to untreated Ti6Al4V using a Student’s unpaired <span class="html-italic">t</span>-test (* = α < 0.05, **** = α < 0.0001).</p> "> Figure 4
<p>DBD treatment effect on cell response. In vitro assessment of bMSCs cultured on argon plasma-treated and untreated Ti6Al4V surfaces. Total DNA content measured at 7 days of culture (<b>A</b>). ELISA quantification of osteoblast maturation markers osteocalcin (<b>B</b>) and osteopontin (<b>C</b>), and paracrine signaling factors osteoprotegerin (<b>D</b>) and vascular endothelial growth factor (<b>E</b>) in response to argon plasma-treated surfaces. Immunomodulatory cytokine production of IL-6 (<b>F</b>) and IL-10 (<b>G</b>). Groups are the means of 6 independent cultures/variables, with error bars representing SEM. Factor production in the conditioned media was normalized to total DNA, and stats were determined using a Student’s unpaired <span class="html-italic">t</span>-test. Groups labeled with “*” are statistically significant compared to untreated Ti6Al4V at <span class="html-italic">p</span>-value equal to or less than 0.05.</p> "> Figure 5
<p>Argon treatment effect on surface properties. Implant surface characterization shows increased wettability following treatment with oxygen plasma-based cleaner under vacuum conditions. Sessile water droplet test of Ti6Al4V surface (<b>A</b>) and Ti6Al4V surface treated with UV–plasma cleaner (<b>B</b>). Contact angle measurements of water droplets for treated and untreated surfaces (<b>C</b>); measures were taken at 6 different locations on the implant surface. Optical profilometry measurements of surface micro-roughness (<b>D</b>) and peak-to-valley height (<b>E</b>). X-ray photoelectron spectroscopy to assess concentrations of elements present on the surface (<b>F</b>). Results are the means of 6 measurements taken at different points on 2 surfaces (<span class="html-italic">n</span> = 12), with bars showing SEM. Groups labeled with “*” are statistically significant compared to untreated Ti6Al4V using a Student’s unpaired <span class="html-italic">t</span>-test (* = α < 0.05, **** = α < 0.0001).</p> "> Figure 6
<p>Argon treatment effect on cell response. In vitro assessment of bMSCs cultured on oxygen plasma under vacuum-treated and untreated Ti6Al4V surfaces. Total DNA content measured at 7 days of culture (<b>A</b>). ELISA quantification of osteoblast maturation markers osteocalcin (<b>B</b>) and osteopontin (<b>C</b>), and paracrine signaling factors osteoprotegerin (<b>D</b>) and vascular endothelial growth factor (<b>E</b>) in response to oxygen plasma vacuum-treated surfaces. Immunomodulatory cytokine production of IL-6 (<b>F</b>) and IL-10 (<b>G</b>). Groups are the means of 6 independent cultures/variables, with error bars representing SEM. Factor production in the conditioned media was normalized to total DNA, and stats were determined using a Student’s unpaired <span class="html-italic">t</span>-test. Groups labeled with “*” are statistically significant compared to untreated Ti6Al4V at <span class="html-italic">p</span>-value equal to or less than 0.05.</p> "> Figure 7
<p>Argon and oxygen plasma treatment effect on surface properties of SLA surfaces. Surface characterization of SLA and modSLA surfaces that were treated with argon or oxygen plasma. Contact angle measurements of water droplets for treated and untreated SLA (<b>A</b>) and modSLA (<b>B</b>) surfaces; measures were taken at 6 different locations on the implant surface. Analysis of SLA surface micro-roughness (<b>C</b>) and peak-to-valley height (<b>D</b>) using optical profilometry. Optical profilometry measurements of surface micro-roughness (<b>E</b>) and peak-to-valley height (<b>F</b>) of modSLA-treated and untreated surfaces. X-ray photoelectron spectroscopy to assess concentrations of elements on untreated SLA and modSLA surfaces and plasma-treated SLA surfaces (<b>G</b>). Results are the means of 6 measurements taken at different points on 2 surfaces (<span class="html-italic">n</span> = 12), with bars showing SEM. Groups labeled with “*” are statistically significant compared to untreated SLA at <span class="html-italic">p</span>-value equal to or less than 0.05.</p> "> Figure 8
<p>Argon and oxygen plasma treatment effect on cell response of SLA surfaces. In vitro assessment of bMSCs cultured on SLA surfaces treated with or without plasma and compared to modSLA. Total DNA content measured at 7 days of culture (<b>A</b>). ELISA quantification of osteoblast maturation markers osteocalcin (<b>B</b>) and osteopontin (<b>C</b>), paracrine signaling factor osteoprotegerin (<b>D</b>), and immunomodulatory cytokines Il-6 (<b>E</b>) and Il-10 (<b>F</b>) in response to SLA surfaces that were treated with either argon or oxygen plasma cleaner and compared to modSLA surfaces. Groups are the means of 6 independent cultures/variables with error bars representing SEM. Factor production in the conditioned media was normalized to total DNA, and stats were determined by ANOVA with Tukey post-test. Groups labeled with “*” are statistically significant compared to SLA at <span class="html-italic">p</span>-value equal to or less than 0.05. Groups labeled with a “#” are statistically significant compared to SLA-AR at <span class="html-italic">p</span>-value equal to or less than 0.05. Groups labeled with a “<span>$</span>” are statistically significant compared to SLA-O<sub>2</sub> at <span class="html-italic">p</span>-value equal to or less than 0.05.</p> "> Figure 9
<p>Argon plasma treatment effect on cell response of SLA and modSLA surfaces. In vitro assessment of bMSCs cultured on SLA and modSLA surfaces and treated with argon plasma. Total DNA content (<b>A</b>) and production of osteogenic markers osteocalcin (<b>B</b>), osteopontin (<b>C</b>), and osteoprotegerin (<b>D</b>) were measured. Production of cytokines Il-6 (<b>E</b>) and Il-10 (<b>F</b>) were measured. Groups are means of 6 independent cultures/variables, with error bars representing SEM. Factor production in the conditioned media was normalized to total DNA and stats were determined by ANOVA with Tukey post-test. Groups labeled with “*” are statistically significant compared to SLA at <span class="html-italic">p</span>-value equal to or less than 0.05. Groups labeled with a “#” are statistically significant compared to SLA AR at <span class="html-italic">p</span>-value equal to or less than 0.05. Groups labeled with a “<span>$</span>” are statistically significant compared to mSLA at <span class="html-italic">p</span>-value equal to or less than 0.05.</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Surface Manufacturing
2.2. UV Light Treatment
2.3. Dielectric Barrier Discharge Plasma Cleaning
2.4. Argon and Oxygen Plasma
2.5. Surface Characterization
2.5.1. Scanning Electron Microscopy
2.5.2. Contact Angle Analysis
2.5.3. Roughness Analysis
2.5.4. Chemical Analysis
2.6. Cell Culture
2.7. Cellular Response
2.8. Statistical Analysis
3. Results
3.1. Effect of UV–Plasma Treatment
3.1.1. Surface Properties
3.1.2. BMSC Response
3.2. Effect of DBD Plasma
3.2.1. Surface Properties
3.2.2. Cell Response
3.3. Effect of Argon Plasma
3.3.1. Ti6Al4V Surface Properties
3.3.2. Cell Response to Ti6Al4V Surfaces
3.4. Effect of Argon and Oxygen Plasmas on Ti Surfaces
3.4.1. Surface Properties
3.4.2. Cell Responses to SLA-O2 and SLA-AR
3.4.3. Cell Response to Plasma-Treated modSLA
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Beschnidt, S.M.; Cacaci, C.; Dedeoglu, K.; Hildebrand, D.; Hulla, H.; Iglhaut, G.; Krennmair, G.; Schlee, M.; Sipos, P.; Stricker, A.; et al. Implant success and survival rates in daily dental practice: 5-year results of a non-interventional study using CAMLOG SCREW-LINE implants with or without platform-switching abutments. Int. J. Implant. Dent. 2018, 4, 33. [Google Scholar] [CrossRef] [PubMed]
- Donos, N.; Calciolari, E. Dental implants in patients affected by systemic diseases. Br. Dent. J. 2014, 217, 425–430. [Google Scholar] [CrossRef] [PubMed]
- Terheyden, H.; Lang, N.P.; Bierbaum, S.; Stadlinger, B. Osseointegration-communication of cells. Clin. Oral. Implants. Res. 2012, 23, 1127–1135. [Google Scholar] [CrossRef] [PubMed]
- Zhao, G.; Schwartz, Z.; Wieland, M.; Rupp, F.; Geis-Gerstorfer, J.; Cochran, D.L.; Boyan, B.D. High surface energy enhances cell response to titanium substrate microstructure. J. Biomed. Mater. Res.-Part A 2005, 74, 49–58. [Google Scholar] [CrossRef] [PubMed]
- Lotz, E.M.; Berger, M.B.; Schwartz, Z.; Boyan, B.D. Regulation of osteoclasts by osteoblast lineage cells depends on titanium implant surface properties. Acta Biomater. 2018, 68, 296–307. [Google Scholar] [CrossRef]
- Zhang, Z.; Xie, Y.; Pan, H.; Huang, L.; Zheng, X. Influence of patterned titanium coatings on polarization of macrophage and osteogenic differentiation of bone marrow stem cells. J. Biomater. Appl. 2018, 32, 977–986. [Google Scholar] [CrossRef] [PubMed]
- Diez-Escudero, A.; Andersson, B.; Carlsson, E.; Recker, B.; Link, H.; Järhult, J.D.; Hailer, N.P. 3D-printed porous Ti6Al4V alloys with silver coating combine osteocompatibility and antimicrobial properties. Biomater. Adv. 2022, 133, 112629. [Google Scholar] [CrossRef]
- Masaki, C.; Schneider, G.B.; Zaharias, R.; Seabold, D.; Stanford, C. Effects of implant surface microtopography on osteoblast gene expression. Clin. Oral. Implants Res. 2005, 16, 650–656. [Google Scholar] [CrossRef]
- Li, R.; Li, S.; Zhang, Y.; Jin, D.; Lin, Z.; Tao, X.; Chen, T.; Zheng, L.; Zhang, Z.; Wu, Q. Titanium surfaces with biomimetic topography and copper incorporation to modulate behaviors of stem cells and oral bacteria. Front. Bioeng. Biotechnol. 2023, 11, 1223339. [Google Scholar] [CrossRef]
- Berger, M.B.; Bosh, K.B.; Cohen, D.J.; Boyan, B.D.; Schwartz, Z. Benchtop plasma treatment of titanium surfaces enhances cell response. Dent. Mater. 2021, 37, 690–700. [Google Scholar] [CrossRef]
- Lee, H.; Jeon, H.J.; Jung, A.; Kim, J.; Kim, J.Y.; Lee, S.H.; Kim, H.; Yeom, M.S.; Choe, W.; Gweon, B.; et al. Improvement of osseointegration efficacy of titanium implant through plasma surface treatment. Biomed. Eng. Lett. 2022, 12, 421–432. [Google Scholar] [CrossRef] [PubMed]
- Lang, N.P.; Salvi, G.E.; Huynh-Ba, G.; Ivanovski, S.; Donos, N.; Bosshardt, D.D. Early osseointegration to hydrophilic and hydrophobic implant surfaces in humans. Clin. Oral. Implants Res. 2011, 22, 349–356. [Google Scholar] [CrossRef] [PubMed]
- Schwarz, F.; Wieland, M.; Schwartz, Z.; Zhao, G.; Rupp, F.; Geis-Gerstorfer, J.; Schedle, A.; Broggini, N.; Bornstein, M.M.; Buser, D.; et al. Potential of chemically modified hydrophilic surface characteristics to support tissue integration of titanium dental implants. J. Biomed. Mater. Res.-Part. B Appl. Biomater. 2009, 88, 544–557. [Google Scholar] [CrossRef] [PubMed]
- Nevins, M.; Chen, C.Y.; Parma-Benfenati, S.; Kim, D.M. Gas Plasma Treatment Improves Titanium Dental Implant Osseointegration-A Preclinical In Vivo Experimental Study. Bioengineering 2023, 10, 1181. [Google Scholar] [CrossRef] [PubMed]
- Henningsen, A.; Precht, C.; Karnatz, N.; Bibiza, E.; Yan, M.; Guo, L.; Gosau, M.; Smeets, R. Osseointegration of titanium implants after surface treatment with ultraviolet light or cold atmospheric plasma in vivo. Int. J. Oral. Implantol. 2023, 16, 197–208. [Google Scholar]
- Wu, C.; Yang, M.; Ma, K.; Zhang, Q.; Bai, N.; Liu, Y. Improvement implant osseointegration through nonthermal Ar/O2 plasma. Dent. Mater. J. 2023, 42, 461–468. [Google Scholar] [CrossRef] [PubMed]
- Metavarayuth, K.; Villarreal, E.; Wang, H.; Wang, Q. Surface topography and free energy regulate osteogenesis of stem cells: Effects of shape-controlled gold nanoparticles. Biomater. Transl. 2021, 2, 165–173. [Google Scholar] [CrossRef] [PubMed]
- Gentleman, M.M.; Gentleman, E. The role of surface free energy in osteoblast–biomaterial interactions. Int. Mat. Rev. 2014, 59, 417–429. [Google Scholar] [CrossRef]
- Duske, K.; Koban, I.; Kindel, E.; Schröder, K.; Nebe, B.; Holtfreter, B.; Jablonowski, L.; Weltmann, K.D.; Kocher, T. Atmospheric plasma enhances wettability and cell spreading on dental implant metals. J. Clin. Periodontol. 2012, 39, 400–407. [Google Scholar] [CrossRef] [PubMed]
- Canullo, L.; Genova, T.; Wang, H.-L.; Carossa, S.; Mussano, F. Plasma of argon increases cell attachment and bacterial decontamination on different implant surfaces. Int. J. Oral. Maxillofac. Implants 2017, 32, 1315–1323. [Google Scholar] [CrossRef]
- Zhang, W.-S.; Liu, Y.; Shao, S.-Y.; Shu, C.-Q.; Zhou, Y.-H.; Zhang, S.-M.; Qiu, J. Surface characteristics and in vitro biocompatibility of titanium preserved in a vitamin C-containing saline storage solution. J. Mater. Sci. Mater. Med. 2024, 35, 3. [Google Scholar] [CrossRef] [PubMed]
- Raghavendra, S.; Wood, M.C.; Taylor, T.D. Early wound healing around endosseous implants: A review of the literature. Int. J. Oral. Maxillofac. Implants 2005, 20, 425–431. [Google Scholar] [CrossRef] [PubMed]
- Sánchez-Puetate, J.C.; Silva, B.L.G.; Pinotti, F.E.; Marcantonio, C.C.; de Oliveira, G.J.P.L.; Junior, E.M.; Marcantonio, R.A.C. Assessing bone formation on hydrophilic and hydrophobic implant surfaces in a murine model treated with bisphosphonates. Clin. Oral. Investig. 2024, 28, 89. [Google Scholar] [CrossRef] [PubMed]
- Park, J.H.; Olivares-Navarrete, R.; Baier, R.E.; Meyer, A.E.; Tannenbaum, R.; Boyan, B.D.; Schwartz, Z. Effect of cleaning and sterilization on titanium implant surface properties and cellular response. Acta Biomater. 2012, 8, 1966–1975. [Google Scholar] [CrossRef] [PubMed]
- Recek, N. Biocompatibility of plasma-treated polymeric implants. Materials 2019, 12, 240. [Google Scholar] [CrossRef] [PubMed]
- Shun’ko, E.V.; Belkin, V.S. Cleaning properties of atomic oxygen excited to metastable state 2 s2 2 p4 (S1 0). J. Appl. Phys. 2007, 102, 083304. [Google Scholar] [CrossRef]
- Berger, M.B.; Cohen, D.J.; Bosh, K.B.; Kapitanov, M.; Slosar, P.J.; Levit, M.M.; Gallagher, M.; Rawlinson, J.J.; Schwartz, Z.; Boyan, B.D. Bone marrow stromal cells generate an osteoinductive microenvironment when cultured on titanium-aluminum-vanadium substrates with biomimetic multiscale surface roughness. Biomed. Mater. 2023, 18, 035001. [Google Scholar] [CrossRef] [PubMed]
- Berger, M.B.; Cohen, D.J.; Levit, M.M.; Puetzer, J.L.; Boyan, B.D.; Schwartz, Z. Hydrophilic implants generated using a low-cost dielectric barrier discharge plasma device at the time of placement exhibit increased osseointegration in an animal pre-clinical study: An effect that is sex-dependent. Dent. Mater. 2022, 38, 632–645. [Google Scholar] [CrossRef] [PubMed]
- Gittens, R.A.; Olivares-Navarrete, R.; Cheng, A.; Anderson, D.M.; McLachlan, T.; Stephan, I.; Geis-Gerstorfer, J.; Sandhage, K.H.; Fedorov, A.G.; Rupp, F.; et al. The roles of titanium surface micro/nanotopography and wettability on the differential response of human osteoblast lineage cells. Acta Biomater. 2013, 9, 6268–6277. [Google Scholar] [CrossRef]
- Rupp, F.; Scheideler, L.; Olshanska, N.; de Wild, M.; Wieland, M.; Geis-Gerstorfer, J. Enhancing surface free energy and hydrophilicity through chemical modification of microstructured titanium implant surfaces. J. Biomed. Mater. Res. A 2006, 76, 323–334. [Google Scholar] [CrossRef]
- Meiron, T.S.; Marmur, A.; Saguy, I.S. Contact angle measurement on rough surfaces. J. Colloid. Interface Sci. 2004, 274, 637–644. [Google Scholar] [CrossRef] [PubMed]
- Krzywicka, M.; Szymańska, J.; Tofil, S.; Malm, A.; Grzegorczyk, A. Surface Properties of Ti6Al7Nb Alloy: Surface Free Energy and Bacteria Adhesion. J. Funct. Biomater. 2022, 13, 26. [Google Scholar] [CrossRef] [PubMed]
- Puzas, V.M.V.; Carter, L.N.; Schröder, C.; Colavita, P.E.; Hoey, D.A.; Webber, M.A.; Addison, O.; Shepherd, D.E.T.; Attallah, M.M.; Grover, L.M.; et al. Surface Free Energy Dominates the Biological Interactions of Postprocessed Additively Manufactured Ti-6Al-4V. ACS Biomater. Sci. Eng. 2022, 8, 4311–4326. [Google Scholar] [CrossRef] [PubMed]
- Lotz, E.M.; Olivares-Navarrete, R.; Berner, S.; Boyan, B.D.; Schwartz, Z. Osteogenic response of human MSCs and osteoblasts to hydrophilic and hydrophobic nanostructured titanium implant surfaces. J. Biomed. Mater. Res. A 2016, 104, 3137–3148. [Google Scholar] [CrossRef]
- Hayashi, R.; Takao, S.; Komasa, S.; Sekino, T.; Kusumoto, T.; Maekawa, K. Effects of Argon Gas Plasma Treatment on Biocompatibility of Nanostructured Titanium. Int. J. Mol. Sci. 2023, 25, 149. [Google Scholar] [CrossRef]
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Jacobs, T.W.; Dillon, J.T.; Cohen, D.J.; Boyan, B.D.; Schwartz, Z. Different Methods to Modify the Hydrophilicity of Titanium Implants with Biomimetic Surface Topography to Induce Variable Responses in Bone Marrow Stromal Cells. Biomimetics 2024, 9, 227. https://doi.org/10.3390/biomimetics9040227
Jacobs TW, Dillon JT, Cohen DJ, Boyan BD, Schwartz Z. Different Methods to Modify the Hydrophilicity of Titanium Implants with Biomimetic Surface Topography to Induce Variable Responses in Bone Marrow Stromal Cells. Biomimetics. 2024; 9(4):227. https://doi.org/10.3390/biomimetics9040227
Chicago/Turabian StyleJacobs, Thomas W., Jonathan T. Dillon, David J. Cohen, Barbara D. Boyan, and Zvi Schwartz. 2024. "Different Methods to Modify the Hydrophilicity of Titanium Implants with Biomimetic Surface Topography to Induce Variable Responses in Bone Marrow Stromal Cells" Biomimetics 9, no. 4: 227. https://doi.org/10.3390/biomimetics9040227