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Effect of citric acid concentration and etching duration on the surface roughness of prepared human tooth enamel: an in vitro study

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1. Title Title of document Effect of citric acid concentration and etching duration on the surface roughness of prepared human tooth enamel: an in vitro study
2. Creator Author's name, affiliation, country Stanislav Yu. Maksyukov; Rostov State Medical University; Russian Federation
2. Creator Author's name, affiliation, country Ekaterina S. Maksyukova; Rostov State Medical University; Russian Federation
2. Creator Author's name, affiliation, country Olga N. Risovannaya; Kuban State Medical University; Russian Federation
3. Subject Discipline(s)
3. Subject Keyword(s) citric acid; etching; tooth enamel
4. Description Abstract

BACKGROUND: Citric acid, commonly encountered through the consumption of fruits and juices, is a frequent factor affecting teeth in daily life. It contributes to enamel dissolution, exposing the underlying dentin in the crown region and potentially leading to significant tooth surface loss. On the other hand, acid etching is widely used in restorative dentistry to enhance the adhesion of restorative materials to tooth surfaces. This process partially dissolves and demineralizes the enamel’s inorganic matrix, creating micropores and microgrooves that increase surface roughness. Pre-etching of enamel improves the bond strength of resin-based adhesives compared to non-etched enamel. Orthodontists also employ acid etching to create retention surfaces, which enhances bracket adhesion but simultaneously weakens adjacent enamel prisms, increases permeability, promotes biofilm adhesion, and reduces enamel’s protective properties.

AIM: To evaluate the effect of citric acid exposure time and etching duration on the surface properties of dental hard tissues, including enamel and dentin.

MATERIALS AND METHODS: Three enamel samples were obtained from a maxillary molar extracted from a 21-year-old male patient. The tooth was embedded in epoxy resin for easier sectioning. Sectioning parameters on the IsoMet 4000 (Buehler, Switzerland) were set to a disk rotation speed of 2500 rpm and a feed rate of 10 mm/min. Three distinct areas were selected for analysis. Each sample was embedded in epoxy resin again for subsequent grinding and polishing, which was performed simultaneously for all three samples using the MetaServ 250 (Buehler, Switzerland). The prepared enamel surfaces were examined using an optical microscope (Stemi 305; Carl Zeiss, Germany) equipped with a color video camera (Axiocam 105; Carl Zeiss, Germany) under reflected light. The samples were etched as follows: Sample 1 with 0.5 wt.% citric acid (pH = 2.3), Sample 2 with 3 wt.% citric acid (pH = 1.86), and Sample 3 with 5 wt.% citric acid (pH = 1.71). The pH was measured using Cobra4 Mobile-Link (PHYWE, Germany). Atomic force microscopy of enamel surface topography before and after etching was performed using the NANOEDUCATOR microscope (NT-MDT, Russia) with a tungsten probe operating in non-contact mode. Surface roughness (Ra) was measured at each etching stage for each sample using the Gwyddion software (Czech Metrology Institute, Czech Republic).

RESULTS: Optical microscopy provided images of enamel, dentin, and the dentinoenamel junction, confirming the healthy condition of the examined samples. The findings suggest that 5 wt.% citric acid is more suitable for controlled etching than 37% phosphoric acid, as prolonged exposure to phosphoric acid can cause severe enamel damage. AFM analysis demonstrated the surface characteristics of each sample before and after etching at different time intervals. A linear increase in surface roughness was observed in all samples after 45 s of etching.

CONCLUSION: For achieving significant surface roughness and visualizing enamel prisms within a reasonable timeframe, etching with 5 wt.% citric acid is recommended. However, further studies on the adhesive properties of enamel post-etching with citric acid are necessary for clinical implementation.

5. Publisher Organizing agency, location Eco-Vector
6. Contributor Sponsor(s) Government of the Russian Federation (14.Z50.31.0046)
7. Date (DD-MM-YYYY) 01.03.2025
8. Type Status & genre Peer-reviewed Article
8. Type Type Research Article
9. Format File format PDF (Rus), PDF (Rus), PDF (Rus),
10. Identifier Uniform Resource Identifier https://rjdentistry.com/1728-2802/article/view/635284
10. Identifier Digital Object Identifier (DOI) 10.17816/dent635284
11. Source Title; vol., no. (year) Russian Journal of Dentistry; Vol 29, No 1 (2025)
12. Language English=en ru
13. Relation Supp. Files Fig. 1. Selected surface areas for atomic force microscopy scanning: a — sample 1; b — sample 2; c — sample 3. © Eco-Vector, 2025. (324KB)
Fig. 2. AFM images of surfaces before etching (a — sample 1, b — sample 2, c — sample 3) and after 1 s of etching (d — sample 1, e — sample 2, f — sample 3). The images do not reflect the details of the microstructure. AFM — atomic force microscopy. © Eco-Vector, 2025. (1MB)
Fig. 3. AFM images of surfaces after 15-second etching of samples (a — sample 1, b — sample 2, c — sample 3) and after 30-second etching (d — sample 1, e — sample 2, f — sample 3). Qualitative change in surface microgeometry. AFM — atomic force microscopy. © Eco-Vector, 2025. (1MB)
Fig. 4. AFM images of sample surfaces after 45 s of etching (a — sample 1, b — sample 2, c — sample 3) and after 60 s of etching (d — sample 1, e — sample 2, f — sample 3). AFM — atomic force microscopy. © Eco-Vector, 2025. (834KB)
Fig. 5. Surface profiles of the samples after each etching step: 1 — without etching, 2 — after etching for 1 s, 3 — for 15 s, 4 — for 30 s, 5 — for 45 s, 6 — for 60 s. © Eco-Vector, 2025. (560KB)
Fig. 6. High-resolution image of sample 1 after 60 s of etching. © Eco-Vector, 2025. (92KB)
Fig. 7. Horizontal surface profile of sample 1 after 60 s of etching (with high resolution). © Eco-Vector, 2025. (63KB)
Fig. 8. Dependence of the surface roughness Ra on the etching time in different directions (1 — horizontal, 2 — vertical, 3 — diagonal) for: а — sample 1, b — sample 2, c — sample 3. © Eco-Vector, 2025. (338KB)
Fig. 9. Dependence of the absolute value of the surface roughness of Raabsol on the etching time for all samples. The number on the graph corresponds to the sample number. © Eco-Vector, 2025. (107KB)
14. Coverage Geo-spatial location, chronological period, research sample (gender, age, etc.)
15. Rights Copyright and permissions Copyright (c) 2025 Eco-Vector
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