Effect of sandblasted large grit acid etching and ultraviolet-c photofunctionalization of titanium on osteoblast cell viability: an in vitro study

Andreas Tjandra, Kwartarini Murdiastuti, Ahmad Kusumaatmaja, Heni Susilowati

Abstract


Introduction: Sandblasted Large Grit Acid Etching (SLA) is a widely used titanium implant surface modification technique that enhances osseointegration. Ultraviolet C (UV-C) photofunctionalization is a method that modifies the physicochemical properties of titanium surfaces by removing hydrocarbon contaminants and increasing hydrophilicity. However, the combined effects of these modification techniques on MC3T3-E1 osteoblast cell viability require further investigation. Therefore, this study aimed to analyze the effect of SLA treatment and its interaction with UV-C irradiation on MC3T3-E1 osteoblast cell viability on titanium disks. Methods: This in vitro experimental study used titanium disks (10 mm in diameter and 1 mm in thickness), which were divided into SLA-treated and non-SLA groups. Each group was further subdivided according to UV-C irradiation (90 W, 254 nm) into four subgroups: non-UV (control), 10 minutes, 30 minutes, and 48 hours (n = 4 per subgroup). MC3T3-E1 osteoblasts (1.2×104 cells/well) were seeded onto disk surfaces and cultured for 24 hours. Cell viability was assessed using the MTT assay. Data normality was evaluated using the Shapiro-Wilk test, homogeneity using the Brown-Forsythe test, and differences between groups using two-way ANOVA, with statistical significance set at p <0.05. Results: Two-way ANOVA revealed that UV irradiation had no significant effect on cell viability (F=2.388; p=0.093). Similarly, SLA treatment showed no significant effect (F= 1.679; p = 0.207), and no significant interaction was observed between SLA treatment and UV-C irradiation (F=0.041; p=0.988). The highest mean cell viability was observed in the non-SLA group exposed to UV-C irradiation for 10 minutes (88.53 ± 25.22), whereas the lowest mean cell viability was found in the SLA group without UV-C irradiation (45.55 ± 16.29). Conclusion: Neither UV-C irradiation nor SLA treatment significantly affected MC3T3-E1 osteoblast cell viability, and no significant interaction was observed between the two treatments.


Keywords


Acid etching, dental; ultraviolet rays; osteoblasts; cell survival; titanium

Full Text:

PDF

References


Tu C, Wang G, Hu Z, Wang S, Yan Q, Liu X. Burden of oral disorders, 1990–2019: estimates from the Global Burden of Disease Study 2019. Arch Med Sci. 2023;19(4):930-940. https://doi.org/10.5114/aoms/165962

Tuikampee S, Chaijareenont P, Rungsiyakull P, Yavirach A. Titanium Surface Modification Techniques to Enhance Osteoblasts and Bone Formation for Dental Implants: A Narrative Review on Current Advances. Metals (Basel). 2024;14(5):515. https://doi.org/10.3390/met14050515

Kido D, Komatsu K, Suzumura T, Matsuura T, Cheng J, Kim J, Park W, Ogawa T. Influence of Surface Contaminants and Hydrocarbon Pellicle on the Results of Wettability Measurements of Titanium. Int J Mol Sci. 2023;24(19):14688. https://doi.org/10.3390/ijms241914688

Rupp F, Liang L, Geis-Gerstorfer J, Scheideler L, Hüttig F. Surface characteristics of dental implants: A review. Dent Mater. 2017;34(1):40-57. https://doi.org/10.1016/j.dental.2017.09.007

Hanawa T. Titanium-Tissue Interface Reaction and Its Control With Surface Treatment. Front Bioeng Biotechnol. 2019;7:170. https://doi.org/10.3389/fbioe.2019.00170

Houshmand B, Rezaei Esfahroodi Z, Behnamghader A, Mohammadreza S, Azizi A, Ramezani K. Evaluation of UV photofunctionalization effect on ultrastructural properties of SLA titanium disks: An in vitro study. J Adv Periodontol Implant Dent. 2023;15(2):117-122. https://doi.org/10.34172/japid.2023.015

Taniyama T, Saruta J, Mohammadzadeh Rezaei N, Nakhaei K, Ghassemi A, Hirota M, Okubo T, Ikeda T, Sugita Y, Hasegawa M, Ogawa T. UV-Photofunctionalization of Titanium Promotes Mechanical Anchorage in A Rat Osteoporosis Model. Int J Mol Sci. 2020;21(4):1235. https://doi.org/10.3390/ijms21041235

Nicholson JW. Titanium Alloys for Dental Implants: A Review. Prosthesis. 2020;2(2):100-116. https://doi.org/10.3390/prosthesis2020011

Yin C, Zhang T, Wei Q, Cai H, Cheng Y, Tian Y, Leng H, Wang C, Feng S, Liu Z. Surface treatment of 3D printed porous Ti6Al4V implants by ultraviolet photofunctionalization for improved osseointegration. Bioact Mater. 2022;7:26-38. https://doi.org/10.1016/j.bioactmat.2021.05.043

Huang Y, Zhang H, Chen Z, Wang Y, Yang X, Yu H. Improvement in Osseointegration of Titanium Dental Implants After Exposure to Ultraviolet-C Light for Varied Durations: An Experimental Study in Beagle Dogs. J Oral Maxillofac Surg. 2022;80(8):1389-1397. https://doi.org/10.1016/j.joms.2022.04.013

Park G, Matsuura T, Komatsu K, Ogawa T. Optimizing implant osseointegration, soft tissue responses, and bacterial inhibition: A comprehensive narrative review on the multifaceted approach of the UV photofunctionalization of titanium. J Prosthodont Res. 2025;69(2):136-152. https://doi.org/10.2186/jpr.JPR_D_24_00086

Izumiya M, Haniu M, Ueda K, Ishida H, Ma C, Ideta H, Sobajima A, Ueshiba K, Uemura T, Saito N, Haniu H. Evaluation of MC3T3-E1 Cell Osteogenesis in Different Cell Culture Media. Int J Mol Sci. 2021;22(14):7752. https://doi.org/10.3390/ijms22147752

Wilkesmann S, Westhauser F, Fellenberg J. Combined Fluorescence-Based in Vitro Assay for the Simultaneous Detection of Cell Viability and Alkaline Phosphatase Activity during Osteogenic Differentiation of Osteoblast Precursor Cells. Methods Protoc. 2020;3(2):30. https://doi.org/10.3390/mps3020030

Präbst K, Engelhardt H, Ringgeler S, Hübner H. Basic colorimetric proliferation assays: MTT, WST, and Resazurin. Methods Mol Biol. 2017;1601:1-17. https://doi.org/10.1007/978-1-4939-6960-9_1

Ihwah A, Deoranto P, Wijana S, Dewi IA. Comparative study between Federer and Gomez method for number of replication in complete randomized design using simulation: study of Areca Palm (Areca catechu) as organic waste for producing handicraft paper. IOP Conf Ser Earth Environ Sci. 2018;131(1):012049. https://doi.org/10.1088/1755-1315/131/1/012049

Ghasemi M, Turnbull T, Sebastian S, Kempson I. The MTT Assay: Utility, Limitations, Pitfalls, and Interpretation in Bulk and Single-Cell Analysis. Int J Mol Sci. 2021;22(23):12827. https://doi.org/10.3390/ijms222312827

Jeon C, Oh KC, Park KH, Moon HS. Effects of ultraviolet treatment and alendronate immersion on osteoblast-like cells and human gingival fibroblasts cultured on titanium surfaces. Sci Rep. 2019;9(1):2581. https://doi.org/10.1038/s41598-019-39355-3

Razali M, Ngeow WC, Omar RA, Chai WL. An integrated overview of ultraviolet technology for reversing titanium dental implant degradation: Mechanism of reaction and effectivity. Appl Sci. 2020;10(5):1654. https://doi.org/10.3390/app10051654

Sanchez-Perez A, Cano-Millá N, Moya Villaescusa MJ, Montoya Carralero JM, Navarro Cuellar C. Effect of Photofunctionalization with 6 W or 85 W UVC on the Degree of Wettability of RBM Titanium in Relation to the Irradiation Time. Appl Sci. 2021;11(12):5427. https://doi.org/10.3390/app11125427

Horikawa H, Yui T, Nakanishi Y, Hirose Y, Kado T, Nezu T, Oh H, Ochi M. Storage of titanium dental implants in ozone nanobubble water retards biological aging and enhances osseointegration: an in vivo study. Materials (Basel). 2025;18(13):3156. https://doi.org/10.3390/ma18133156

Cho YD, Kim WJ, Kim S, Ku Y, Ryoo HM. Surface topography of titanium affects their osteogenic potential through dna methylation. Int J Mol Sci. 2021;22(5):2406. https://doi.org/10.3390/ijms22052406

Matsuura T, Komatsu K, Cheng J, Park G, Ogawa T. Beyond microroughness: novel approaches to navigate osteoblast activity on implant surfaces. Int J Implant Dent. 2024;10(1):35. https://doi.org/10.1186/s40729-024-00554-x

Lee JB, Jo YH, Choi JY, Seol YJ, Lee YM, Ku Y, Rhyu IC, Yeo ISL. The effect of ultraviolet photofunctionalization on a titanium dental implant with machined surface: An in vitro and in vivo study. Materials (Basel). 2019;12(13):2078. https://doi.org/10.3390/ma12132078

Osman MA, Alamoush RA, Kushnerev E, Seymour KG, Shawcross S, Yates JM. In-vitro phenotypic response of human osteoblasts to different degrees of titanium surface roughness. Dent J (Basel). 2022;10(8):140. https://doi.org/10.3390/dj10080140

Roy M, Corti A, Dorocka-Bobkowska B, Pompella A. positive effects of uv-photofunctionalization of titanium oxide surfaces on the survival and differentiation of osteogenic precursor cells—an in vitro study. J Funct Biomater. 2022;13(4):265. https://doi.org/10.3390/jfb13040265

Guo L, Zou Z, Smeets R, Kluwe L, Hartjen P, Cacaci C, Gosau M, Henningsen A. Time dependency of non-thermal oxygen plasma and ultraviolet irradiation on cellular attachment and mrna expression of growth factors in osteoblasts on titanium and zirconia surfaces. Int J Mol Sci. 2020;21(22):8598. https://doi.org/10.3390/ijms21228598

Suzumura T, Matsuura T, Komatsu K, Ogawa T. A novel high-energy vacuum ultraviolet light photofunctionalization approach for decomposing organic molecules around titanium. Int J Mol Sci. 2023;24(3):1978. https://doi.org/10.3390/ijms24031978

Agidigbi TS, Kim C. Reactive oxygen species in osteoclast differentiation and possible pharmaceutical targets of ros-mediated osteoclast diseases. Int J Mol Sci. 2019;20(14):3576. https://doi.org/10.3390/ijms20143576

Abdulhameed EA, Al-Rawi NH, Omar M, Khalifa N, Samsudin ABR. Titanium dioxide dental implants surfaces related oxidative stress in bone remodeling: a systematic review. PeerJ. 2022;10:e12951. https://doi.org/10.7717/peerj.12951

Ramakrishnan R, Daly AC. Revisiting ISO 10993-5 In Vitro Cytotoxicity Standard Tests for Evaluation of Extracellular Matrix-Based Biomaterials. Int J Biomater. 2026;2026:7395612. https://doi.org/10.1155/ijbm/7395612




DOI: https://doi.org/10.24198/pjd.vol38no2.70560

Refbacks

  • There are currently no refbacks.


 

Creative Commons License All publications by the Universitas Padjadjaran [e-ISSN: 2549-6212, p-ISSN: 1979-0201] are licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.This license requires that reusers give credit to the creator. It allows reusers to distribute, remix, adapt, and build upon the material in any medium or format, for noncommercial purposes only.

Visitor Stat