Optimization of Anodization Time for TiO2 Nanotubes and their Antibiofilm Activity against Staphylococcus aureus and Pseudomonas aeruginosa
J. Environ. Nanotechnol., Volume 15, No 1 (March 2026) pp. 262-269
Abstract
This study presents a novel investigation into the synthesis and optimization of titanium dioxide nanotube (TiO2) nanotubes (TNTs) fabricated via electrochemical anodization at varying times (1, 2, and 3 hours) under a constant 20V potential. Comprehensive characterization by FESEM, XRD, and EDX confirmed the formation of highly ordered anatase-phase nanotubes, with the 2-hour anodization condition yielding the most uniform morphology and superior antibacterial efficiency. Anti-biofilm activity was assessed against Staphylococcus aureus and Pseudomonas aeruginosa using Microtitre Plate Assay, showing significant inhibition rates of 45.7% and 61.18% at 500 ug/mL, respectively, compared to controls. These findings underscore the optimized time’s critical role in enhancing TiO2 nanotubes antibiofilm properties. Furthermore, the study highlights potential improvements through incorporating silver and graphene nanocomposites to further boost antimicrobial performance, marking a significant advancement in biomedical implant surface modification.
Full Text
Reference
Abish, V. J., Raja, D. H. and Davidson, D. J., Na-deposited TiO2 nanotubes for photo electrochemical water splitting with enhanced activity–A novel approach, Mater. Lett., 362, 136210(2024).
https://doi.org/10.1016/j.matlet.2024.136210
Abisharani, J. M., Devikala, S., Kumar, R. D., Arthanareeswari, M. and Kamaraj, P., Green synthesis of TiO2 nanoparticles using Cucurbita pepo seeds extract, Mater. Today: Proce., 14, 302-307(2019).
https://doi.org/10.1016/j.matpr.2019.04.151
Alipal, J., Lee, T. C., Koshy, P., Abdullah, H. Z. and Idris, M. I., Evolution of anodised titanium for implant applications, Heliyon, 7(7), e07408 (2021).
https://doi.org/10.1016/j.heliyon.2021.e07408
Amalanathan, M., Aravind, M., Ahmed, N., Mary, M. S. M., Velusamy, P., Kumaresubitha, T., Noreen, R. and Ali, S., The influence of activated carbon annealing temperature on sunlight-driven photocatalytic dye degradation and biological activity, Inorg. Chem. Commun., 146, 110149 (2022).
https://doi.org/10.1016/j.inoche.2022.110149
Aravind, M., Ahmad, A., Ahmad, I., Amalanathan, M., Naseem, K., Mary, S. M. M. and Zuber, M., Critical green routing synthesis of silver NPs using jasmine flower extract for biological activities and photocatalytical degradation of methylene blue, J. Environ. Chem. Eng., 9(1), 104877(2021a).
https://doi.org/10.1016/j.jece.2020.104877
Aravind, M., Amalanathan, M. and Mary, S. M. M., Synthesis of TiO2 nanoparticles by chemical and green synthesis methods and their multifaceted properties, SN Appl. Sci., 3, 409 (2021b).
https://doi.org/10.1007/s42452-021-04281-5
Ayal, A. K., Effect of Anodization Duration in the TiO2 Nanotubes Formation on Ti Foil and Photoelectrochemical Properties of TiO2 Nanotubes, J. Sci., 29(3), 77–81 (2019).
https://doi.org/10.23851/mjs.v29i3.640
Bhadra, C. U., Davidson, D. J. and Raja, D. H., Fabrication of titanium oxide nanotubes by varying the anodization time, Mater. Today: Proce., 33, 2711-2715(2020).
https://doi.org/10.1016/j.matpr.2020.01.455
Brudzisz, A. M., Gizinski, D. and Stępniowski, W. J., Incorporation of Ions into Nanostructured Anodic Oxides—Mechanism and Functionalities, Mol., 26(21), 16378 (2021).
https://doi.org/10.3390/molecules26216378
Chen, S., Zhang, S., Tan, Z. and Zhang, S., Effect of Two-step Anodization on structure of TiO2 Nanotube Arrays, Adv. Func. Mater. Proce. China Mater. Conf., 18, 947-955(2017).
https://doi.org/10.1007/978-981-13-0110-0_102
Chernozem, R. V., Surmeneva, M. A. and Surmenev, R. A., Influence of Anodization Time and Voltage on the Parameters of TiO2 Nanotubes, IOP Conf. Ser.: Mater. Sci. Eng., 116, 012025 (2016).
https://doi.org/10.1088/1757-899X/116/1/012025
Farrell, D. J., File, T. M. and Jenkins, S. G., Prevalence and antibacterial susceptibility of mef (A)-positive macrolide-resistant Streptococcus pneumoniae over 4 years (2000 to 2004) of the PROTEKT US Study, J. Clinic. Microbio., 45(2), 290-293(2007).
https://doi.org/10.1128/JCM.01653-06
Garg, P., Attri, P., Sharma, R., Chauhan, M. and Chaudhary, G. R., Advances and Perspective on Antimicrobial Nanomaterials for Biomedical Applications, Front. Nanotech., 4, 898411 (2022).
https://doi.org/10.3389/fnano.2022.898411
Han, S. C., Doh, J. M., Yoon, J. K., Kim, G. H., Byun, J. Y., Han, S. H., Hong, K. T. and Kwun, S. I., Highly ordered self-organized TiO2 nanotube arrays prepared by a multi-step anodic oxidation process, Met. Mater. Int., 15, 493–499 (2009).
https://doi.org/10.1007/s12540-009-0493-x
Jedi-soltanabadi, Z., Pishkar, N. and Ghoranneviss, M., Enhanced physical properties of the anodic TiO2 nanotubes via proper anodization time, J. Theorey Appl. Phy., 12(2), 135-139(2018).
https://doi.org/10.1007/s40094-018-0290-3
Kahlmeter, G., An international survey of the antimicrobial susceptibility of pathogens from uncomplicated urinary tract infections: the ECO·SENS Project, J. Antimicrob. Chemother., 51, 69–76 (2003).
https://doi.org/10.1093/jac/dkg028
Kunrath, M. F., Hübler, R., Shinkai, R. and Teixeira, E. R., TiO2 nanotubes as an antibacterial nanotextured surface with potential for dental implants: Systematic review and meta-analysis, Mater. Sci. Eng. C, 40(6), 907-920(2024).
https://doi.org/10.1016/j.dental.2024.04.009
Lai, C. W. and Sreekantan, S., Photoelectrochemical Performance of Smooth TiO2 Nanotube Arrays: Effect of Anodization Temperature and Cleaning Methods, Int. J. Photoenergy, 2012(1), 356943 (2012).
https://doi.org/10.1155/2012/356943
Mahmud, A. H., Habiballah, A. S. and Jani, A. M. M., The Effect of Applied Voltage and Anodisation Time on Anodized Aluminum Oxide Nanostructures, Mater. Sci. Forum, Trans Tech Publ., 819, 103–108 (2015).
https://doi.org/10.4028/www.scientific.net/MSF.819.103
Mazzarolo, A., Lee, K., Vicenzo, A. and Schmuki, P., Anodic TiO2 nanotubes: Influence of top morphology on their photocatalytic performance, Electrochem. Commun., 22, 162–165 (2012).
https://doi.org/10.1016/j.elecom.2012.05.037
Meyer, E., Jonas, D., Schwab, F., Rueden, H., Gastmeier, P. and Daschner, F. D., Design of a surveillance system of antibiotic use and bacterial resistance in German intensive care units (SARI), Infect., 31(4), 208-215(2003).
https://doi.org/10.1007/s15010-003-3201-7
Meyer, E., Schwab, F., Gastmeier, P., Rueden, H. and Daschner, F. D., Surveillance of antimicrobial use and antimicrobial resistance in German intensive care units (SARI): a summary of the data from 2001 through 2004, Infect., 34(6), 303-309(2006).
https://doi.org/10.1007/s15010-006-6619-x
Mohan, L., Dennis, C., Padmapriya, N., Anandan, C., and Rajendran, N. J. M. T. C., Effect of electrolyte temperature and anodization time on formation of TiO2 nanotubes for biomedical applications, Mater. Today Commun., 23, 101103(2020).
https://doi.org/10.1016/j.mtcomm.2020.101103
Montakhab, E., Rashchi, F. and Sheibani, S., Modification and photocatalytic activity of open channel TiO2 nanotubes array synthesized by anodization process, Appl. Surf. Sci., 534, 147581 (2020).
https://doi.org/10.1016/j.apsusc.2020.147581.
Opolot, E. E., Zhang, L., Abogodiri, S. O., Kumar, R., and Truong, V. T., Synergistic antibacterial activity and inhibition of TiO2 nanotube array against multi-drug-resistant bacteria, Front. Microbiol., 15,616649(2024).
https://doi.org/10.3389/fmicb.2024.618849
O'Toole, G. A., Microtiter dish biofilm formation assay, J. Visual. Exp., (47), 2437(2011).
Pollán, M., Pérez-Gómez, B., Pastor-Barriuso, R., Oteo, J., Hernán, M. A., Pérez-Olmeda, P., Jose, L. S., Aurora, F. G., Isreal, C., Nerea, F. D. L., Marta, M., Francisco, R. C., Mariano, M. Paloma, M. A., Joose, L. P., Juan, F. M. M., Faustino, B. and Raquel, Y., 396(10250), 535–544 (2020).
https://doi.org/10.1016/S0140-6736(20)31483-5
Rahimpour, N., Mollavali, M. and Rashidi, F., A Comprehensive Study on Synthesis Parameters of TiO2 Nanotube Arrays: The Effects on Morphology and Photocatalytic Degradation of Para-NitroPhenol, Phy. Chem, Res., 9(2), 261-279(2021).
https://doi.org/10.22036/pcr.2021.257129.1853
Raja, D. H. and Arsha, A., Optical and morphological studies of Titania nanotubes and their characterizations, Next Mater., 4, 100215 (2024).
https://doi.org/10.1016/j.nxmate.2024.100215
Rathi, A. K., Kmentová, H., Naldoni, A., Goswami, A., Gawande, M. B., Varma, R. S. and Zbořil, R., Significant enhancement of photoactivity in hybrid TiO2/g-C3N4 nanorod catalysts modified with Cu–Ni-based nanostructures, ACS Appl. Nano Mater., 1(6), 2526-2535(2018).
https://doi.org/10.1021/acsanm.8b00078
Regonini, D., Satka, A., Jaroenworaluck, A., Allsopp, D. W. E., Bowen, C. R. and Stevens, R., Factors influencing surface morphology of anodized TiO2 nanotubes, Electrochim. Acta, 74, 244–253 (2012).
https://doi.org/10.1016/j.electacta.2012.04.076
Rojviroon, T., Rojviroon, O. and Sirivithayapakorn, S., Photocatalytic decolourisation of dyes using TiO2 thin film photocatalysts, Surf. Eng., 32(8), 562-569(2016).
https://doi.org/10.1179/1743294415Y.0000000096
Sun, M., Fang, Y., Kong, Y., Sun, S., Yu, Z. and Umar, A., Graphitic carbon nitride (g-C3N4) coated titanium oxide nanotube arrays with enhanced photo-electrochemical performance, Dalton Trans., 45, 12702–12709 (2016).
https://doi.org/10.1039/C6DT02071B
Surya, B., Ahmad, D. N. A., Sakti, H. H. and Sahban, H., Land Use Change, Spatial Interaction, and Sustainable Development in the Metropolitan Urban Areas, Land, 9(3), 95 (2020).
https://doi.org/10.3390/land9030095
Torres, Á. I. P., Souza, R. M., Chino-Ulloa, A., Ruiz-Trabolsi, P. A., Tadeo-Rosas, R., Carrera-Espinoza, R. and Hernández, S. E., Crystals, 13(7), 1059 (2023).
