Biofilm formation of mixed Candida albicans and methicillin-sensitive Staphylococcus aureus and surface properties of a 3D-printed denture base resin under different printing parameters
DOI:
https://doi.org/10.1590/1678-7765-2025-0526Keywords:
Denture bases, Biofilms, Printing, Three-DimensionalAbstract
Objective To evaluate the formation of mixed-species biofilms of Candida albicans and methicillin-sensitive Staphylococcus aureus (MSSA) on the surface of a 3D-printed denture base resin, as well as its surface properties, under varying printing parameters. Methodology Discs (n=40 per group, 10×1.2 mm) of a denture base resin (priZma 3D Bio Denture) were fabricated using two 3D-printers—Liquid Crystal Display (LCD) and Digital Light Processing (DLP)—at three different angles (0°, 45°, or 90°). Surface roughness was measured using a digital profilometer and expressed as Ra (µm). For surface energy (SE) analysis, contact angles were measured using a tensiometer. Discs were incubated at 37 °C for 90 minutes and 48 hours to enable biofilm formation using C. albicans and MSSA inocula. Cell viability was assessed by colony-forming unit (CFU/mL) counts, and metabolic activity was evaluated using the XTT assay (absorbance). Microbial counts and XTT results were analyzed by three-way ANOVA (printer type, printing angle, incubation period). Surface roughness was analyzed by two-way ANOVA (printer type, printing angle), with Tukey’s test and a significance level of 0.05. Results For both CFU/mL and XTT assays, incubation period was the only significant factor (p<0.001 and p=0.006, respectively), while other factors and interactions were not statistically significant (p>0.05). Surface roughness was significantly influenced by printer type, printing angle, and their interaction (p=0.027). The LCD 0° and LCD 90° groups produced smoother surfaces compared with LCD 45° (p=0.002), which showed similar values to all DLP groups regardless of angle (p>0.05). The DLP printer did not show significant roughness variations across the tested angles (p>0.05). The LCD groups presented numerically lower SE values compared to the DLP groups. Conclusion The LCD system performs better than DLP in reducing surface roughness at 0° and 90°. Moreover, the analyzed factors did not significantly affect microbial adhesion or the formation of mixed-species biofilms.
Downloads
References
1- Alharbi N, Wismeijer D, Osman RB. Additive manufacturing techniques in prosthodontics: where do we currently stand? A critical review. Int J Prosthodont. 2017;30(5):474-84. doi: 10.11607/ijp.5079
» https://doi.org/10.11607/ijp.5079
2- Aati S, Akram Z, Shrestha B, Patel J, Shih B, Shearston K, et al. Effect of post-curing light exposure time on the physico-mechanical properties and cytotoxicity of 3D-printed denture base material. Dent Mater. 2022;38(1):57-67. doi: 10.1016/j.dental.2021.10.011
» https://doi.org/10.1016/j.dental.2021.10.011
3- Altarazi A, Haider J, Alhotan A, Silikas N, Devlin H. Assessing the physical and mechanical properties of 3D printed acrylic material for denture base application. Dent Mat. 2022;38(12):1841-54. doi: 10.1016/j.dental.2022.09.006
» https://doi.org/10.1016/j.dental.2022.09.006
4- Chen H, Cheng DH, Huang SC, Lin YM. Comparison of flexural properties and cytotoxicity of interim materials printed from mono-LCD and DLP 3D printers. J Prosthet Dent. 2021;126(5):703-8. doi: 10.1016/j.prosdent.2020.09.003
» https://doi.org/10.1016/j.prosdent.2020.09.003
5- Turanoglu OF, Talay Cevlik E, Vural C. Investigation of adhesion status of Candida species to the surface of resin materials produced at different angles with additive manufacturing. BMC Oral Health. 2024 Jun 27;24(1):738. doi: 10.1186/s12903-024-04505-1
» https://doi.org/10.1186/s12903-024-04505-1
6- Silva MD, Carmo Viotto HE, Moisés LD, Coelho SR, Souza RF, Pero AC. Stainability of 3D-printed resins for denture base and artificial teeth. BMC Oral Health. 2025;25(1):1-9. doi: 10.1186/s12903-024-05367-3
» https://doi.org/10.1186/s12903-024-05367-3
7- Lee HE, Alauddin MS, Mohd Ghazali MI, Said Z, Mohamad Zol S. Effect of different vat polymerization techniques on mechanical and biological properties of 3D-printed denture base. Polymers. 2023;15(6):1463. doi: 10.3390/polym15061463
» https://doi.org/10.3390/polym15061463
8- Hada T, Kanazawa M, Iwaki M, Arakida T, Soeda Y, Katheng A, et al. Effect of printing direction on the accuracy of 3D-printed dentures using stereolithography technology. Materials (Basel). 2020;13(15):3405. doi: 10.3390/ma13153405
» https://doi.org/10.3390/ma13153405
9- Li P, Fernandez PK, Spintzyk S, Schmidt F, Beuer F, Unkovskiy A. Effect of additive manufacturing method and build angle on surface characteristics and Candida albicans adhesion to 3D printed denture base polymers. J Dent. 2022;116:103889. doi: 10.1016/j.jdent.2021.103889
» https://doi.org/10.1016/j.jdent.2021.103889
10- Unkovskiy A, Bui PH, Schille C, Geis-Gerstorfer J, Huettig F, Spintzyk S. Objects build orientation, positioning, and curing influence dimensional accuracy and flexural properties of stereolithographically printed resin. Dent Mater. 2018;34(12):e324-e333. doi: 10.1016/j.dental.2018.09.011
» https://doi.org/10.1016/j.dental.2018.09.011
11- Al-Dulaijan YA, Alsulaimi L, Alotaibi R, Alboainain A, Alalawi H, Alshehri S, et al. Comparative evaluation of surface roughness and hardness of 3D printed resins. Materials (Basel). 2022;15(19):6822. doi: 10.3390/ma15196822
» https://doi.org/10.3390/ma15196822
12- Goodacre BJ, Goodacre CJ. Additive manufacturing for complete denture fabrication: a narrative review. J Prosthod. 2022;31(S1):47-51. doi: 10.1111/jopr.13426
» https://doi.org/10.1111/jopr.13426
13- Radford DR, Sweet SP, Challacombe SJ, Walter JD. Adherence of Candida albicans to denture-base materials with different surface finishes. J Dent. 1998;26(7):577-83. doi: 10.1016/s0300-5712(97)00034-1
» https://doi.org/10.1016/s0300-5712(97)00034-1
14- Alghazzawi TF. Advancements in CAD/CAM technology: options for practical implementation. J Prosthod Res. 2016;60(2):72-84. doi: 10.1016/j.jpor.2016.01.003
» https://doi.org/10.1016/j.jpor.2016.01.003
15- Nair RG, Samaranayake LP. The effect of oral commensal bacteria on candidal adhesion to denture acrylic surfaces. An in vitro study. APMIS. 1996;104(5):339-49. doi: 10.1111/j.1699-0463.1996.tb00725.x
16- Akpan A, Morgan R. Oral candidiasis. Postgrad Med J. 2002;78(922):455-9. doi: 10.1136/pmj.78.922.45
» https://doi.org/10.1136/pmj.78.922.45
17- Chandra J, Mukherjee PK, Leidich SD, Faddoul FF, Hoyer LL, Douglas LJ, et al. Antifungal resistance of candidal biofilms formed on denture acrylic in vitro. J Dent Res. 2001;80(3):903-8. doi: 10.1177/00220345010800031101
» https://doi.org/10.1177/00220345010800031101
18- Morales DK, Hogan DA. Candida albicans interactions with bacteria in the context of human health and disease. PLoS pathogens. 2010;6(4):e1000886. doi: 10.1371/journal.ppat.1000886
» https://doi.org/10.1371/journal.ppat.1000886
19- Sánchez-Vargas LO, Estrada-Barraza D, Pozos-Guillen AJ, Rivas-Caceres R. Biofilm formation by oral clinical isolates of Candida species. Arch Oral Biology. 2013;58(10):1318-26. doi: 10.1016/j.archoralbio.2013.06.006
» https://doi.org/10.1016/j.archoralbio.2013.06.006
20- Baena-Monroy T, Moreno-Maldonado V, Franco-Martínez F, Aldape-Barrios B, Quindós G, Sánchez-Vargas LO. Candida albicans, Staphylococcus aureus and Streptococcus mutans colonization in patients wearing dental prosthesis. Med Oral Patol Oral Cir Bucal. 2005;10:E27-39. Spanish.
21- Ponde NO, Lortal L, Ramage G, Naglik JR, Richardson JP. Candida albicans biofilms and polymicrobial interactions. Crit Rev Microbiol. 2021;47(1):91-111. doi: 10.1080/1040841X.2020.1843400
» https://doi.org/10.1080/1040841X.2020.1843400
22- De Foggi CC, Machado AL, Zamperini CA, Fernandes D, Wady AF, Vergani CE. Effect of surface roughness on the hydrophobicity of a denture-base acrylic resin and Candida albicans colonization. J Investig Clin Dent. 2016;7(2):141-8. doi: 10.1111/jicd.12125
» https://doi.org/10.1111/jicd.12125
23- Shim JS, Kim JE, Jeong SH, Choi YJ, Ryu JJ. Printing accuracy, mechanical properties, surface characteristics, and microbial adhesion of 3D-printed resins with various printing orientations. J Prosthet Dent. 2020;124(4):468-75. doi: 10.1016/j.prosdent.2019.05.034
» https://doi.org/10.1016/j.prosdent.2019.05.034
24- Janssen D, De Palma R, Verlaak S, Heremans P, Dehaen W. Static solvent contact angle measurements, surface free energy and wettability determination of various self-assembled monolayers on silicon dioxide. Thin Solid Films. 2006;515(4):1433-8.
25- Behroozi F. A fresh look at the young-laplace equation and its many applications in hydrostatics. The Physics Teacher. 2022;60(5):358-61.
26- Yacob N, Ahmad NA, Safii SH, Yunus N, Razak FA. Is microbial adhesion affected by the build orientation of a 3-dimensionally printed denture base resin? J Prosthet Dent. 2023;130(1):131.e1-131.e7. doi: 10.1016/j.prosdent.2023.04.017
» https://doi.org/10.1016/j.prosdent.2023.04.017
27- Nunes TS, Silva MD, Coelho SR, Viotto HE, Pero AC. Effectiveness of disinfectant solutions associated or not with brushing on the biofilm control of a 3D printed-denture base resin. J Appl Oral Sci. 2023;31:e20230104. doi: 10.1590/1678-7757-2023-0104
» https://doi.org/10.1590/1678-7757-2023-0104
28- Paleari AG, Marra J, Pero AC, Rodriguez LS, Ruvolo-Filho A, Compagnoni MA. Effect of incorporation of 2-tert-butylaminoethyl methacrylate on flexural strength of a denture base acrylic resin. J Appl Oral Sci. 2011;19(3):195-9. doi: 10.1590/s1678-77572011000300003
» https://doi.org/10.1590/s1678-77572011000300003
29- Rodriguez LS, Paleari AG, Giro G, Oliveira NM Junior, Pero AC, Compagnoni MA. Chemical characterization and flexural strength of a denture base acrylic resin with monomer 2-tert-butylaminoethyl methacrylate. J Prosthodont. 2013;22(4):292-7. doi: 10.1111/j.1532-849X.2012.00942.x
30- Song F, Koo H, Ren D. Effects of material properties on bacterial adhesion and biofilm formation. J Dent Res. 2015;94(8):1027-34. doi: 10.1177/0022034515587690.
31- Minagi S, Miyake Y, Inagaki K, Tsuru H, Suginaka H. Hydrophobic interaction in Candida albicans and Candida tropicalis adherence to various denture base resin materials. Infect Immun. 1985;47(1):11-4. doi: 10.1128/iai.47.1.11-14.1985
» https://doi.org/10.1128/iai.47.1.11-14.1985
32- Cerca N, Pier GB, Vilanova M, Oliveira R, Azeredo J. Quantitative analysis of adhesion and biofilm formation on hydrophilic and hydrophobic surfaces of clinical isolates of Staphylococcus epidermidis Res Microbiol. 2005;156(4):506-14. doi: 10.1016/j.resmic.2005.01.007
» https://doi.org/10.1016/j.resmic.2005.01.007
33- Gad MM, Abualsaud R, Khan SQ. Hydrophobicity of denture base resins: a systematic review and meta-analysis. J Int Soc Prev Community Dent. 2022;12(2):139-59. doi: 10.4103/jispcd.JISPCD_213_21
» https://doi.org/10.4103/jispcd.JISPCD_213_21
34- Lee MJ, Kim MJ, Oh SH, Kwon JS. Novel dental poly (methyl methacrylate) containing phytoncide for antifungal effect and inhibition of oral multispecies biofilm. Materials (Basel). 2020;13(2):371. doi: 10.3390/ma13020371
» https://doi.org/10.3390/ma13020371
35- Revilla-León M, Meyers MJ, Zandinejad A, Özcan M. A review on chemical composition, mechanical properties, and manufacturing workflow of additively manufactured current polymers for interim dental restorations. J Esthet Restor Dent. 2019;31(1):51-7. doi: 10.1111/jerd.12438
» https://doi.org/10.1111/jerd.12438
36- Pereira-Cenci T, Cury AA, Cenci MS, Rodrigues-Garcia RC. In vitro Candida colonization on acrylic resins and denture liners: influence of surface free energy, roughness, saliva, and adhering bacteria. Int J Prosthodont. 2007;20(3):308-10.
37- Dutra D, Pereira G, Kantorski KZ, Valandro LF, Zanatta FB. Does finishing and polishing of restorative materials affect bacterial adhesion and biofilm formation? A systematic review. Oper Dent. 2018;43(1):E37-E52. doi: 10.2341/17-073-L
» https://doi.org/10.2341/17-073-L
38- Zissis AJ, Polyzois GL, Yannikakis SA, Harrison A. Roughness of denture materials: a comparative study. Int J Prosthodont. 2000;13(2):136-40.
39- Gulati M, Nobile CJ. Candida albicans biofilms: development, regulation, and molecular mechanisms. Microbes Infect. 2016;18(5):310-21. doi: 10.1016/j.micinf.2016.01.002
» https://doi.org/10.1016/j.micinf.2016.01.002
40- Uppuluri P, Chaturvedi AK, Srinivasan A, Banerjee M, Ramasubramaniam AK, Köhler JR, et al. Dispersion as an important step in the Candida albicans biofilm developmental cycle. PLoS Pathog. 2010;6(3):e1000828. doi: 10.1371/journal.ppat.1000828
Downloads
Published
Issue
Section
License
Copyright (c) 2026 Larianne S. Moisés, Hamile Emanuella do Carmo Viotto, Sabrina Romão Gonçalves Coelho, Danny Omar Mendoza Marin, Raphael Freitas Souza, Ana Carolina Pero

This work is licensed under a Creative Commons Attribution 4.0 International License.
Todo o conteúdo do periódico, exceto onde está identificado, está licenciado sob uma Licença Creative Commons do tipo atribuição CC-BY.