Ethylenediaminetetraacetic Acid Conditioning Research Articles

Fibroblast Viability Through Mechanical and Chemical Root Surface Modifications in Periodontal Healing: An In Vitro Comparative Study.

The wound-healing process incorporates a spectrum of periodontal therapeutic interventions that strive to restore the health and function of the periodontium. Fibroblasts play pivotal roles in tissue repair and regeneration. Extensive research has been focused on mechanical and chemical root surface modifications to enhance fibroblast adhesion, which is crucial for successful wound healing. This study aimed to assess the combined efficacy of mechanical and chemical root surface modifications in promoting fibroblast viability to root surfaces affected by periodontitis in comparison to chemical modifications alone. Root samples were collected from healthy individuals and those with advanced periodontitis. The specimens were prepared, and the experimental groups were categorized based on the type of surface modification with mechanical and/or chemical materials, including hyaluronic acid (HA), ethylenediaminetetraacetic acid(EDTA), enamel matrix derivatives (EMD), and EDTA/EMD. Fibroblasts were seeded onto previously treated root samples. Cell adhesion was assessed using a viability assay. Fibroblast viability was significantly higher on root surfaces treated with chemical agents than on those treated with mechanical and chemical modifications. Long-duration EDTA and short-duration EMD treatments were significantly effective in enhancing cell viability. EDTA/EMD surface treatments resulted in significantly higher cell viability in all groups compared to the periodontitis root surfaces. EDTA, EMD, and their combined application can potentially ameliorate periodontitis-induced surface structural impairments. Mechanical surface debridement can significantly affect the effectiveness of EDTA and EMD root conditioning agents.

Effect of chitosan irrigant solutions on the release of bioactive proteins from root dentin.

To identify the effect of two chitosan solutions on the release of root dentin matrix proteins and to describe the chemical changes observed following conditioning with chelating agents. The release of dentin sialoprotein (DSP), transforming growth factor-beta 1 (TGF-β1), vascular endothelial growth factor (VEGF), and platelet-derived growth factor-BB (PDGF-BB) with different chelating agents, including ethylenediaminetetraacetic acid (EDTA), chitosan solution (CS), and nanoparticulate chitosan (CSnp), was investigated. DSP was quantified using an enzyme-linked immunosorbent assay (ELISA). TGF-β1, VEGF, and PDGF-BB were quantified using a cytokine bead panel (CBA). Raman spectroscopy was performed to identify surface chemical changes. Statistical analysis was performed using Kruskal-Wallis test with Mann-Whitney-Wilcoxon rank-sum test (p < 0.05). TGF-β1, VEGF, and DSP solubilized in all irrigants tested. CSnp showed the highest concentration of DSP. PDGF-BB did not exceed the detection limits. Raman spectroscopy revealed a decrease in the phosphate and carbonate peaks, representing the chelating effect of EDTA, CS, and CSnp. Additionally, CSnp showed the greatest preservation of the amide I and III content. Proteins can be released from dentin via EDTA, CS, and CSnp conditioning. Raman spectroscopic revealed changes in the inorganic content of the root dentin after chelation. Furthermore, use of CSnp facilitated a preservation of the organic content. Chelation allows the release of proteins, justifying the use of chelating agents in regenerative endodontics. The chitosan-dentin matrix interaction also promotes the protection of the organic content as an additional benefit to its protein releasing effect.

Influence of ethylenediaminetetraacetic acid on regenerative endodontics: A systematic review.

The effects of ethylenediaminetetraacetic acid (EDTA) on regenerative endodontic procedures (REPs) are controversial, because, despite releasing growth factors from dentine, some studies show negative effects on cell behaviour. The aim of the study was to investigate the influence of the use of EDTA in REP on the growth factors' release, cell behaviour and tissue regeneration. A systematic search was conducted (PubMed/Medline, Scopus, Cochrane Library, Web of Science, Embase, OpenGrey and reference lists) up to February 2021. Only in vivo and in vitro studies evaluating the effects of EDTA on the biological factors of dentine, pulp/periapical tissues and cell behaviour were eligible. Studies without a control group or available full text were excluded. The growth factors' release was the primary outcome. Risk of bias in the in vitro and in vivo studies was performed according to Joanna Briggs Institute's Checklist and SYRCLE's RoB tool, respectively. Of the 1848 articles retrieved, 36 were selected. Amongst these, 32 were in vitro, three animal studies and one with both models. The EDTA concentrations ranged from 3% to 15%, at different times. Regarding growth factors' release (17 studies), 15 studies found significant transforming growth factor (TGF)-β release after dentine conditioning with EDTA, and most found no influence on vascular endothelial growth factor release. Regarding cell behaviour (26 studies), eight studies showed no influence of EDTA-treated dentine on cell viability, whereas, five, nine and six studies showed higher cell migration, adhesion and differentiation respectively. No influence of EDTA conditioning was observed in animal studies. In vitro studies had a low risk of bias, whereas animal studies had high risk of bias. Meta-analysis was unfeasible. This review found that EDTA increased TGF-β release and improved cell activity. However, well-designed histological analyses using immature teeth models are needed. High-quality in vitro evidence suggests that EDTA-treated dentine positively influences TGF-β release, cell migration, attachment and differentiation; further research to evaluate its influence on tissue regeneration is necessary due to low methodological quality of the animal studies.

LPCGF and EDTA conditioning of the root surface promotes the adhesion, growth, migration and differentiation of periodontal ligament cells.

Liquid-phase concentrated growth factor (LPCGF), a new-generation platelet concentrate, may potently stimulate human periodontal ligament (PDL) cells. This study assessed the effectiveness of ethylenediaminetetraacetic acid (EDTA) and/or LPCGF on periodontally diseased root surfaces through their effects on PDL cells. Dentin blocks prepared from periodontal teeth were divided into four groups and treated as follows: group I, scaling and root planning (SRP); group II, SRP + EDTA; group III, SRP + LPCGF; and group IV, SRP+EDTA + LPCGF. PDL cells were cultured on dentin blocks, and LPCGF-induced biological effects were evaluated by migration and cell adhesion/proliferation assays. Furthermore, PDL cell differentiation was assessed by real-time polymerase chain reaction (PCR). Significantly more adherent cells were observed in the EDTA, LPCGF and combination treatment groups than in the control group. Root conditioning with EDTA and/or LPCGF enhanced cell proliferation and migration more than SRP did. Compared with the control group, the combined treatment group exhibited significant upregulation of cell differentiation-related genes. Electron microscopy of the tooth surface revealed removal of the smear layer and exposed dentin holes in the EDTA-treated group but not in the control group. EDTA and LPCGF application to periodontitis-affected root surfaces forms a surface suitable for cell attachment, growth, migration and differentiation. Thus, LPCGF is promising in clinical periodontics applications. Further studies to support these results are necessary.

Effect of root conditioning agents hyaluronic acid, EDTA and chlorhexidine on the attachment of human gingival fibroblasts to healthy root surface

BackgroundThe gold standard treatment of periodontal diseases is scaling and root planing (SRP). Different adjunctive root conditioning agents such as hyaluronic acid (HA), ethylenediaminetetraacetic acid(EDTA), and chlorhexidine (CHX) have been used with SRP to improve the smear layer removal and the healing of periodontal tissues. The aim of this study was to compare the effect of manual scaling with or without HA, EDTA, or CHX root conditioning on the attachment and viability of human gingival fibroblasts (GF). MethodsFifteen healthy single rooted teeth were co llected and divided randomly into a scaled (n = 12) and non-scaled control group (n = 3). The scaled roots were subdivided equally into four groups; the first group did not receive any chemical treatment, while the remaining groups were treated with the conditioning agents HA or 17% EDTA or 0.2% CHX gel. Gingival fibroblasts were seeded on the top of each root and incubated for 48 h to allow attachment to the roots. The viability of fibroblasts attached to the root surface was assessed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) cell viability assay. ResultsThe cell viability was the highest in the scaled only group (p = 0.0001) while the lowest was in the scaled with EDTA group (p > 0.05). The scaled group was the highest followed by the HA and CHX groups, while the EDTA group showed the lowest mean value. ConclusionSRP remains the superior method for regaining cell attachment to the root surface, leading to better periodontal health, and adjunctive therapies did not enhance the GF attachment to the root surface beyond the effect of SRP. Further studies are needed to investigate the effect of root conditioning agents on periodontally diseased teeth in vitro and compare them in vivo.

Microtensile Bond Strength of Resin-Modified Glass Ionomer Cement to Sound and Artificial Caries-Affected Root Dentin With Different Conditioning.

In this laboratory study, the microtensile bond strengths (μTBS) of resin-modified glass ionomer cement (RM-GIC) to sound and artificial caries-affected bovine root dentin (ACAD) using three different conditioning agents were evaluated after 24 hours and three months. The fractured interface was examined with a scanning electron microscope (SEM). Specimens were created on bovine root dentin that was embedded in epoxy resin. For the ACAD specimens, artificial carious lesions were created. The RM-GIC (Fuji II LC) was applied either directly (no treatment), after application of self conditioner, cavity conditioner, or 17% ethylenediamine tetraacetic acid (EDTA) applied for 60 seconds, on sound dentin and ACAD, then light cured. They were stored in artificial saliva for 24 hours or three months. Following this, the specimens were cut into sticks for the μTBS test, and the failure mode of the debonded specimens was examined by using SEM. Pretest failures were excluded from the statistical analysis of the μTBS values because of their high incidence in some groups. Results showed that the μTBS values were significantly affected by the dentin substrate as well as the conditioning agent. Self conditioner provided the highest and most stable μTBS values, while cavity conditioner showed stable μTBS values on sound dentin. Both self conditioner and cavity conditioner had significantly higher μTBS values than the no treatment groups. EDTA conditioning reduced the μTBS after three months to sound dentin, while it showed 100% pretest failure with ACAD for both storage periods.

A zinc oxide-modified hydroxyapatite-based cement facilitated new crystalline-stoichiometric and amorphous apatite precipitation on dentine.

To evaluate the remineralization ability of two endodontic sealer cements. Mid-coronal dentine surfaces were subjected to: (i) 37% phosphoric acid (PA) or (ii) 0.5mol L-1 ethylenediaminetetraacetic acid (EDTA) conditioning prior to the application of two experimental hydroxyapatite-based cements, containing sodium hydroxide (calcypatite) or zinc oxide oxiapatite respectively. Samples were stored in simulated body fluid for 24h or 21days. Remineralization of the dentine surfaces were studied by Raman spectroscopy (mapping with K-means cluster and hierarchical cluster analysis) was undertaken. Nanoroughness and collagen fibril width measurements were performed with an atomic force microscopy. ANOVA and Student-Newman-Keuls test were performed (α=0.05). Phosphoric acid+oxiapatite promoted both the highest dentine mineralization (P<0.05) and crystallographic maturity at the dentine surface. Noncrystalline amorphous-like apatites were also formed. Dentine treated with PA+calcypatite attained the roughest surface (P<0.05) with minimal fibril width (P<0.05). Cross-linking of collagen only became greater in the group PA+oxiapatite after 21days. The maximum relative mineral concentration and structure of collagen linked to the amide I and ratio amide III/AGEs was obtained after using PA+calcypatite at 21-days time-point (P<0.05). EDTA produced a lower stoichiometric hydroxyapatite (P<0.05) with decreased maturity, at the expense of carbonate band widening, although it favoured the nucleation of carbonated calcium phosphate. Dentine surfaces treated with PA+oxiapatite attained the highest dentine remineralization with both crystalline-stoichiometric and amorphous apatites, at 21days. EDTA conditioning facilitated amorphous-bulk mineral precipitation. The amorphization was more intense after using oxiapatite and provided an ion-rich environment favouring insitu dentine remineralization.

Effect of Different Protocols in Preconditioning With EDTA in Sclerotic Dentin and Enamel Before Universal Adhesives Applied in Self-etch Mode.

The aim of this study was to investigate the effect of different protocols of 17% ethylene diamine tetra-acetic acid (EDTA) conditioning on the etching pattern and immediate bond strength of universal adhesives to enamel and sclerotic dentin. Forty bovine teeth with sclerotic dentin and 20 human third molars were randomly divided into eight groups resulting from the combination of the main factors surface treatment (none, two-minute EDTA conditioning manual application, 30-second EDTA manual application, 30-second EDTA sonic application) and adhesives systems (Scotchbond Universal Adhesive [SBU] and Prime & Bond Elect [PBE]). Resin-dentin and enamel-dentin bond specimens were prepared and tested under the microtensile bond strength (μTBS) and microshear bond strength (μSBS) tests, respectively. The etching pattern produced on the unground enamel and the sclerotic dentin surfaces under the different protocols and adhesive systems was evaluated under scanning electron microscopy. For enamel, only the main factor adhesive was significant (p<0.0001), with SBU showing the highest μSBS. In sclerotic dentin, the lowest mean was observed for the group without EDTA application and the highest mean in the group with EDTA application with the sonic device for 30 seconds. Regardless of the EDTA protocol, the highest means of μTBS were observed for SBU (p<0.05). EDTA conditioning improves the bonding performance of universal adhesives in the self-etch mode on sclerotic dentin, mainly when applied for 30 seconds with the aid of a sonic device. EDTA pretreatment also improves the retentive etching pattern of enamel, but it does not result in higher enamel bond strength.

Ultrasonic activation of irrigants increases growth factor release from human dentine.

Bioactive proteins are sequestered in human dentine and play a decisive role in dental pulp regeneration and repair. They can be released and exposed on the dentine surface by acids, but also chelators, such as ethylenediaminetetraacetic acid (EDTA). The objectives of this study were (i) to evaluate whether ultrasonic activation of irrigants in the root canal will promote growth factor release from dentine and (ii) to collect bioactive proteins in a physiological solution. Human dentine disks underwent irrigation with and without ultrasonic activation. The protocols included treatment by either a single or two consecutive steps with 10% EDTA and phosphate-buffered saline (PBS), where each sample was treated three times. To mimic clinical conditions, selected irrigation regimens were applied to root canals of extracted human teeth after preparation. Amounts of transforming growth factor β1 (TGF-β1) in solution were quantified using enzyme-linked immunosorbent assays. Nonparametric statistical analysis was performed to compare different groups as well as repetitions within a group (Mann-Whitney U test, α=0.05). Additionally, morphological changes of dentine surfaces were visualized by scanning electron microscopy (SEM). TGF-β1 was not detectable after irrigation of dentine with PBS, neither with nor without ultrasonic activation. Irrigation with EDTA released TGF-β1, and ultrasonic activation of EDTA enhanced this effect. However, preceding EDTA conditioning enabled the release of bioactive proteins into PBS solution. Similar results were observed in dentine disks and root canals. Visualization of dentine surfaces after different treatment revealed superficial erosion after ultrasonic activation irrespective of the irrigant solution, but different degrees of exposure of organic substance. Ultrasonic activation enhances growth factor release from human dentine. Bioactive proteins can be isolated in physiological solvents and may act as autologous supplements for regenerative endodontic treatment or pulp tissue engineering. Autologous growth factors from human dentine can advance treatment strategies in dental pulp tissue engineering.

Retentive strength of fiber-reinforced composite posts with composite resin cores: Effect of remaining coronal structure and root canal dentin conditioning protocols

Statement of problemThe prognosis of a fixed dental prosthesis cemented to endodontically treated teeth is primarily determined by the presence of a ferrule on the tooth. Adhesion of the post in the root canal, conditioning methods for the canal and the amount of coronal structure could also be decisive on survival of reconstructions cemented on endodontically treated teeth. PurposeThe purpose of this in vitro study was to test the effect of remaining coronal structure on the retention of airborne-particle abraded fiber-reinforced composite resin posts built up with composite resin cores after the treatment of root canal dentin with different conditioning protocols. Material and methodsOne hundred and fifty extracted human teeth with single root canal space were endodontically treated and divided into 3 groups as follows: group CEJ: the teeth were sectioned at the level of cementoenamel junction (CEJ); group CEJ1: the teeth were sectioned 1 mm above the CEJ; group CEJ2: the teeth were sectioned 2 mm above the CEJ. Each group was further divided into 5 subgroups (n=10 per group) according to the root canal treatments as follows: group C: no conditioning (control); group PH: conditioning with 37% phosphoric acid gel for 15 seconds; group E: conditioning with 17% ethylenediaminetetraacetic acid (EDTA) for 60 seconds; group CHX: conditioning with 2% chlorhexidine (CHX) for 60 seconds; group Q: conditioning with combination of 2% CHX with 17% EDTA and a surfactant solution for 60 seconds. Glass fiber-reinforced composite resin posts were airborne-particle abraded and luted to the root canal dentin with a self-adhesive resin cement (RelyX Unicem). The retentive force was tested by applying a tensile load parallel to the long axis of these posts at a crosshead speed of 2 mm/min. Two-way ANOVA and the Tukey HSD post hoc test were used to analyze the data. ResultsThe highest retention (N) was obtained with the CHX-EDTA conditioned group (374.7 ±29.8) followed by 17% EDTA (367.9 ±33.3) conditioning when 2 mm remaining coronal structure was available. Conditioning with the CHX-EDTA showed comparable retention values to 17% EDTA conditioned groups when 0 or 1 mm coronal structure was present that differed significantly compared to 37% PH and 2% CHX conditioning (P<.05). ConclusionsConditioning root canal either with CHX-EDTA or 17% EDTA delivered superior retention values for fiber-reinforced composite resin posts with composite resin cores that were luted with self-adhesive resin cement to endodontically treated teeth with 2 mm remaining coronal structure.

Effect of EDTA Conditioning and Carbodiimide Pretreatment on the Bonding Performance of All-in-One Self-Etch Adhesives.

Objective. This study evaluated the effect of ethylenediaminetetraacetic acid (EDTA) conditioning and carbodiimide (EDC) pretreatment on the shear bond strength of two all-in-one self-etch adhesives to dentin. Methods. Flat coronal dentin surfaces were prepared on one hundred and sixty extracted human molars. Teeth were randomly divided into eight groups according to two different self-etch adhesives used [G-Bond and OptiBond-All-In-One] and four different surface pretreatments: (a) adhesive applied following manufacturer's instructions; (b) dentin conditioning with 24% EDTA gel prior to application of adhesive; (c) EDC pretreatment followed by application of adhesive; (d) application of EDC on EDTA conditioned dentin surface followed by application of adhesive. Composite restorations were placed in all the samples. Ten samples from each group were subjected to immediate and delayed (6-month storage in artificial saliva) shear bond strength evaluation. Data collected was subjected to statistical analysis using three-way ANOVA and post hoc Tukey's test at a significance level of p < 0.05. Results and Conclusion. EDTA preconditioning as well as EDC pretreatment alone had no significant effect on the immediate and delayed bond strengths of either of the adhesives. However, EDC pretreatment on EDTA conditioned dentin surface resulted in preservation of resin-dentin bond strength of both adhesives with no significant fall over six months.

Impact of oxalate desensitizer combined with ethylene-diamine tetra acetic acid-conditioning on dentin bond strength of one-bottle adhesives during dry bonding

Background:Elimination of water entrapment in hybrid layer during bonding procedure would increase bonding durability.Aims:This study evaluated the effect of oxalate desensitizer (OX) pretreatment on bond strength of three one-bottle adhesives to ethylene-diamine tetra acetic acid (EDTA)-conditioned dentin under dry bonding.Materials and Methods:Three adhesive systems, One-Step Plus (OS), Optibond Solo Plus (OP) and Adper Single Bond (SB) were bonded on dentin surfaces under four bonding conditions: (1) Wet-bonding on acid-etched dentin, (2) wet bonding on EDTA-conditioned dentin, (3) dry bonding on EDTA-conditioned dentin, (4) dry bonding associated with OX on the EDTA-conditioned dentin. After storage and thermo cycling, shear bond strength test was performed. Data were analyzed using two-way analysis of variance and Tukey tests.Results:Wet bonding with EDTA or acid etching showed similar bond strength for test adhesives. Dry bonding with EDTA significantly decreased the bond strength of OS, but it had no effect on the bonding of OP and SB. OX application in the forth bonding condition, in comparison with the third condition, had a negative effect on the bond strength of OP, but not influence on OS and SB.Conclusions:The use of an OX on EDTA-conditioned dentin compromised the bonding efficacy of OS and OP under dry bonding but compatible for SB.

A double‐blind randomized clinical evaluation of enamel matrix derivative proteins for the treatment of proximal class‐II furcation involvements

The aim of the present randomized, double-blind study was to evaluate the clinical response of proximal furcations treated with enamel matrix derivative proteins (EMD). Fifteen patients, each with a pair of contralateral class-II proximal furcation involvements, presenting probing depths (PDs) >/=5 mm and bleeding on probing (BOP) were selected. The patients were randomly assigned to: control group (n=15) - open flap debridement (OFD)+24% ethylenediaminetetraacetic acid (EDTA) conditioning; test group (n=15) - OFD+24% EDTA conditioning+EMD application. Plaque index (PI), BOP, PD, gingival margin position (GMP), relative vertical and horizontal clinical attachment level (RVCAL and RHCAL), vertical and horizontal bone level (VBL and HBL) and furcation closure were evaluated immediately before and 2, 4 and 6 months after the surgeries. At 6 months, the RVCAL gains of the control and test group were 0.39 +/- 1.00 and 0.54 +/- 0.95 mm, while the RHCAL gains were 1.21 +/- 2.28 and 1.36 +/- 1.26 mm (p>0.05). The VBL and HBL gains of the control group were 1.04 +/- 1.12 and 1.00 +/- 1.79 mm, and 0.82 +/- 1.82 and 1.17 +/- 1.38 mm for the test group (p>0.05). In addition, a statistical difference was observed in the number of the remaining class-II furcations between the test and control groups (p<0.05) in this period. It may be concluded that the use of EMD in proximal furcations did not promote a superior reduction in PD or a gain in clinical and osseous attachment levels, but resulted in a higher rate of class-II to class-I furcation conversion.

Shear strength after ethylenediaminetetraacetic acid conditioning of dentin

On the basis of previous studies, it was hypothesized that a chelating agent such as ethylenediaminetetraacetic acid (EDTA) in a saturated aqueous solution (24%)can function as a dentin conditioning agent with exposure times comparable to that of phosphoric acid without compromising shear bond strength. Thirty caries-free human third molars, divided equally between two groups, were used. In group one, four experimental surfaces were prepared on each tooth, and cylindrical copper matrixes with a diameter of 5 mm were attached to the prepared surfaces. The experimental surfaces were then treated with a 24% EDTA gel for 30, 60, 120, or 240 sec, respectively. Dentin was bonded with All Bond 2, after which a flowable composite was added and light-cured. In group two, which served as control, two surfaces were prepared on each tooth. One surface was left unetched, whereas the other side was treated with 24% EDTA-gel for 30 sec. A shear bond strength test was performed at a crosshead speed of 1 mm/min until the composite debonded. There was no statistically significant difference (P < 0.89) between results of the shear bond strength test for the different EDTA conditioning times. The control group showed a significant difference in shear bond strength between untreated surfaces and surfaces conditioned for 30 sec with EDTA. Thus, the results indicate that the duration of EDTA gel conditioning of dentin surfaces need not exceed that of phosphoric acid in clinical practice to obtain an acceptable level of bond strength.