Dentin permeability after pretreatment with titanium tetrafluoride and self-etching or universal adhesive systems

Abstract

Aim

To evaluate dentin permeability after pretreatment with 2.5% aqueous solution of titanium tetrafluoride (TiF₄), followed by a self-etching universal adhesive system.

Methods

Forty dentin discs (1.5 mm thick) were randomly divided into groups according to the application or non-application of a pretreatment, and the type of adhesive system to be tested (two-step self-etching/ Clearfil SE Bond/ Kuraray Medical, or universal adhesive system/ Single Bond Universal/ 3M ESPE). Both sides of the discs were conditioned with 37% phosphoric acid to remove the smear layer. The first hydraulic conductivity measurement (L1) was performed in a permeability machine, under 5 PSI pressure. The samples were sanded again to form a standardized smear layer. The teeth designated for pre-treatment with TiF₄ received the TiF₄ solutions applied actively for 60 seconds, and the adhesive systems were applied according to the manufacturer’s instructions. Then, a new hydraulic conductivity measurement (L2) was performed for the purpose of calculating the hydraulic conductance at a later time, considering the water viscosity and the thickness of the specimen. The percentage (L) of dentin permeability after application of the adhesive system was obtained (L (%) = [(L1-L2) x100] / L1). The Mann-Whitney non-parametric test was applied.

Results

There was no difference between the two adhesive systems, or between the groups with or without pretreatment, as regards dentin permeability (p>0.05).

Conclusion

Pretreatment with 2.5% TiF₄ did not influence dentin permeability, irrespective of the adhesive system used.

Dentin permeability; Dentin-bonding agents; Titanium; Fluorides

Introduction

Advancements in the development of products for adhesive restorative systems has allowed direct restorative procedures to be performed with minimally invasive cavity preparations¹. Although the methods for bonding contemporary adhesive systems to dental substrates are easy to perform, there is always room for improvement in their composition, with the aim of increasing their bond strength to different substrates, and thus increasing the longevity of the restorations².

Dentin has dental tubules filled with circulating dentinal fluid, under constant pulp pressure, which keeps the surface of this substrate permanently moist and makes adhesion difficult³, especially when there are active carious lesions in deep dentin⁴. In self-etching adhesive systems, the tubules remain partially occluded, because of incomplete smear layer removal; hence, the dentin surface is less susceptible to the effects of pulp pressure⁵. Since universal adhesive systems work by way of chemical bonding to the tooth structure, it is questionable whether the tooth permeability resulting from these adhesive systems is lower than it would be with older generation adhesives.

The suggestion has been that some products can be used together with adhesive systems to increase the longevity of the bond to dental tissues, especially dentin, among them chlorhexidine⁶, and some phenolic compounds present in green tea⁷. The aforementioned products have been evaluated for their potential of inhibiting endogenous enzymes, preventing proteolytic activity in collagen fibers, and maintaining the hybrid layer for a longer time. Their ability to increase cross-links between collagen fibrils has also been assessed with a view to promoting increased resistance to degradation by endogenous proteases⁸.

Titanium tetrafluoride (TiF₄) is an inorganic fluoride compound that has also been researched as a dentin pretreatment to increase the longevity of the hybrid layer⁹-¹¹. TiF₄ is usually used in the form of gel, varnish or aqueous solution, applied to the enamel surface to prevent caries lesions, reduce in vitro demineralization¹², and inhibit the progression of caries lesions in situ and in vivo¹³. It has the ability to prevent erosion and abrasion injuries¹⁴, and to reduce the risk of dentin hypersensitivity¹⁵.

When TiF₄ is hydrolyzed, it acquires a low pH rate, at which titanium tends to bind to an oxygen atom of a phosphate group on the tooth surface¹⁶. This leads to the formation of a solid element composed of titanium oxides or organometallic complexes, and a stable, modified, acid-resistant smear layer¹⁷,¹⁸. In this case, occlusion of the dentinal tubules occurs¹⁸, which is capable of reducing dentin permeability¹⁹.

When dentin pretreatment with 2.5% TiF₄ was applied, before or after acid etching, followed by the application of a conventional adhesive system, there were no changes in the bond strength to dentin²⁰, and similar results were reported with self-etching adhesives⁹-¹¹. However, the influence of dentin pretreatment with 2.5% TiF₄ associated with self-etching and universal adhesive systems on dentin permeability have not yet been evaluated. Therefore, the aim of this study was to evaluate dentin permeability after pretreatment with a 2.5% aqueous solution of titanium tetrafluoride (TiF₄), followed by the use of a self-etching of universal adhesive system.

Materials and methods

Sample Preparation

Forty healthy human third molars (approved by the Ethics Committee - CAAE 16187519.6.0000.5374) were used, cleaned with scalpel blades, washed with water, and stored frozen until ready to use. The crowns were sectioned perpendicular to the long axis of the tooth, 1.5 mm above the enamel-cement junction, using a precision saw (Isomet 1000, Buehler, Springfield, VA, USA) under cooling, to obtain the dentin discs²¹. The samples were sanded on both sides with 600 grit sandpaper to ensure uniformity and smoothness of the surfaces²². The occlusal surface of the sample was sanded until it was completely free of enamel. A diamond tip was used to mark the occlusal face that was used to adjust the slice of the permeability machine. The thickness of each sample was checked with a digital caliper (Mitutoyo Sul Americana, MIP/E, Suzano, SP, Brazil). The final thickness of the dentin discs was 1.5 mm.

The discs were rinsed with water and stored in a flask with 5 mL distilled water in an incubator for 24 h. After this period, the slices were removed from the flask and first dried with paper for 15 s, and then with air for 5 s. After this, the smear layer was created by removing the polish with 37% phosphoric acid for 15 s²¹ and washing with water, on both sides for 30 s. Acid etching was performed²³ to open the dentinal tubules and increase dentin permeability, as a way to ensure standardization of the first hydraulic conductivity measurement (L1) (considered 100%), corresponding to the maximum filtration. Afterwards, the occlusal surfaces were polished with 300 grit sandpaper for 30 s to form a standardized smear layer²².

A total of 40 samples were prepared and randomly distributed into four groups (n=10). Then they were dried with absorbent paper for 15 s, and received adhesive treatment on the occlusal surface, as specified for the particular group, according to the manufacturer’s recommendations.

Dentin treatments

The 40 dentin specimens were separated into groups (n = 10): Pretreatment + Clearfil SE Bond, Pretreatment + Single Bond Universal, Clearfil SE Bond, Single Bond Universal. The composition of the adhesive systems is described in Table 1.

Table 1
Adhesive systems evaluated in this study

Materials	Composition	pH	Manufacturer (city, state, country)
Clearfil SE Bond	Primer: HEMA, 10-MDP, hydrophilic aliphatic dimethacrylate, DL- camphorquinone, water, accelerators, dyes Bond: Bis-GMA, HEMA, 10-MDP, hydrophobic aliphatic dimethacrylate, silica, DL-camphorquinone, initiators, accelerators	Bond 1.55 Primer 1.55	Kuraray Medical Inc. (1621 Sakazu, Kurashiki, Okayama, Japan)
Single Bond Universal	HEMA, Bis-GMA, ethanol, water, 10-MDP, silica, copolymer of acrylic and itaconic acid, silane, camphorquinone, dimethylaminobenzoate	2.82	3M ESPE (Sumaré, São Paulo, Brazil)

• Bis-GMA: bisphenol A diglycidyl methacrylate; HEMA: 2-hydroxyethyl methacrylate; 10-MDP: 10-Methacryloyloxydecyl dihydrogen phosphate

The teeth assigned to receive the dentin pre-treatment were brushed with TiF₄ P.A. (Sigma Aldrich, Saint Louis, MO, USA) dissolved in distilled water to a final concentration of 2.5% (w/v; pH 1.0). The solution was actively applied to the dentin surface using a disposable brush (Microbrush Corporation, Grafton, WI, USA) for 60 s. After this, the adhesive systems for each group were applied according to the respective manufacturer’s instructions.

The primer assigned for use with the Clearfil SE Bond adhesive system was actively applied with a microbrush for 20 s, and then air dried for 5 s at a distance of 10 cm. Another microbrush was used to actively apply the adhesive over the primer for 20 s, and air was applied for 5 s immediately afterwards. The Single Bond Universal adhesive system was actively applied for 20 s and received an air spray for 5 s.

The adhesive system was light-cured with a photoactivation device (VALO, Ultradent, UT, USA) for 20 s with a light intensity of 1000 mW/cm². Then, a new hydraulic conductivity measurement was performed (L2).

Permeability Test

A permeability machine (THD, Odeme Dental Research, Luzerna, SC, Brazil) was used under 5 PSI pressure, equivalent to 351.54 cmH₂O²³. The dentin disc was attached to the filtration chamber device, and the machine system was adjusted. The water entered the dentinal tubules and exerted pressure toward the surface. Detachment of the liquid was marked by the difference in location of the air bubble inside the glass microtube of the equipment

Three measurements were performed during continuous movement of the liquid inside the glass microtube, to ensure that the amount of fluid that passed through the sample could be calculate using the following mathematical formula: Q=(ri²l)/ t, where Q (μL / min-1) was the amount of liquid passing through the sample, l(cm) was the linear displacement in the glass capillary, t (min) was the time, and ri (cm²) was the internal measurement of the glass microtube.

The hydraulic conductivity (L) was obtained considering the viscosity of the water and thickness of the constant specimen, L = Q / (AP), where L is the hydraulic conductivity (μL cm-² min-¹ cmH₂O-¹), A (cm²) was the dentin surface area, and P (cmH₂O) was the pressure imposed. The hydraulic conductivity of each dentin disc was evaluated at two time points: initially, after acid etching (L1), and after applying the adhesive system protocol with or without TiF₄ pretreatment (L2).

The percentage of dentin permeability after application of the adhesive system was obtained using the equation below, with each tooth being its own control: L (%) = [(L1-L2) x100] / L1²⁴, where L was the percentage of permeability, L1 was the hydraulic conductance after removal of the smear layer, and L2 was the hydraulic conductivity after application of the adhesive system.

Statistical Analysis

Data distribution was evaluated by the Shapiro-Wilk test. Exploratory analysis indicated that the data did not meet the assumptions of analysis of variance (ANOVA). The Mann Whitney non-parametric test was then applied. The results were summarized and presented with median, minimum and maximum values. All the analyses were performed using the R program, with a significance level of 5%.

Results

There was wide variability (%) in permeability among the dentin discs in the same group (Table 2). No significant differences were found between the two groups studied, or between the groups with or without dentin pretreatment, as regards permeability (p>0.05).

Table 2
Median (%) (minimum and maximum value) dentin permeability according to application of dentin pretreatment and type of adhesive system

Adhesive System	Dentin Pretreatment		p-value
	None	2.5% TiF4 aqueous solution
Single Bond Universal	43.84 (17.68; 81.59)	47.76 (-64.42; 77.32)	0.5967
Clearfil SE Bond	40.26 (-7.57; 71.01)	53.67 (-73.23; 70.84)	0.6501
p-value	0.3258	0.8206

Discussion

The formation of a hybrid layer with greater permeability causes greater degradation that compromises the tooth-restoration interface and dentinal sealing²⁵. Instead, lower permeability of the hybrid layer is desirable because it leads to less elution of resin components²⁶, and minimum sorption and solubility rates?/levels?/. Nevertheless, dentin pretreatment with 2.5% TiF₄ did not influence dentin permeability, irrespective of the association with either of the two adhesive systems.

Use of TiF₄ in an aqueous solution has been shown to form a layer with a vitreous aspect and promote a reduction in dentin permeability when used in concentrations of 0.1, 0.5, 1.0²⁷ and 4%²⁸. This mechanism has been explained by the occlusion of the dentinal tubules¹⁸, which reduced dentin permeability¹⁹, especially when the smear layer was removed before the application of TiF₄²⁸. In contrast, this procedure differed from that of the present study, in which the smear layer was preserved. However, it should be noted that the adhesives used in this study had a mild pH (about 1.55 for Clearfil SE Bond), and ultra-mild (about 2.82 for Single Bond Universal)²⁹. It could be suggested that these pH values caused the adhesives to dilute the vitreous layer, especially considering that the adhesive systems were actively applied (by brushing the dentin)⁹. Although Sen and Büyükyilmaz¹⁷(1998) showed that the smear layer treated with TiF₄ was resistant to treatments with 17% EDTA and 5.25% sodium hypochlorite, their results cannot be compared with those of the present study, since the concentration of TiF₄ was 4%, whereas a 2.5% solution was used in the present study.

The ability of self-etching adhesives to establish a mechanically resistant bond to dentin surfaces treated with an aqueous solution of 2.5% TiF₄ has been reported in previous studies⁹-¹¹,³⁰, despite this pretreatment solution tending to flocculate, and having a higher-than-average particle size³⁰. These adhesive systems have been observed to be capable of penetrating the vitreous layer formed by the application of an aqueous solution of TiF₄. The acidity of the self-etching primer probably caused demineralization of the modified dentin surface layer, leading to the formation of a hybrid layer⁹-¹¹,³⁰. Furthermore, the hydrophilic functional monomer 10-MDP most likely contributed significantly to the chemical bonding to dentin³¹.

Single Bond Universal is a single-bottle adhesive, containing hydrophilic components believed to promote greater permeability, by contributing to the formation of a semi-permeable hybrid layer²⁵. Whereas Clearfil SE Bond adhesive has fewer hydrophilic components when compared with Single Bond. However, both adhesives are self-etching, a type of system that does not require etching with 37% phosphoric acid. Cruz et al.³²(2021) also compared etch-and-rinse vs. self-etch adhesive system application modalities and observed similar results to those found in the present study, with no reduction in dentin permeability for Single Bond Universal or Clearfil SE adhesives, irrespective of the application modality. In the present study, both adhesive systems were used without removing the smear layer, hence favoring a decrease in permeability⁵,³³. Both adhesives also promoted a chemical interaction with hydroxyapatite through the 10-methacryloxidecyl phosphate monomer (10-MDP), thereby favoring a more stable adhesive interface³⁴,³⁵. Thus, the self-etching application mode and the presence of 10-MDP in both adhesive systems may have contributed to their similar permeability level.

The results of the present study suggested that dentin pretreatment with 2.5% TiF₄ did not reduce dentin permeability. However, future studies must be conducted to assess whether this permeability is affected over time, especially in relation to reducing hybrid layer degradation.

In conclusion, the use of 2.5% TiF₄ as a dentin pretreatment, combined with self-etching and universal adhesive systems, did not influence dentin permeability.

Acknowledgements

The authors would like to thank the CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico) for providing grants (registration number 800280/2018-0).

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• Data availability

  Datasets related to this article are available from the corresponding author upon request.

Edited by

Editor: Dr. Altair A. Del Bel Cury

Data availability

Datasets related to this article are available from the corresponding author upon request.

Publication Dates

• Publication in this collection
  09 June 2023
• Date of issue
  2023

History

• Received
  11 Nov 2021
• Accepted
  2 Apr 2022