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      Attachment of human periodontal ligament fibroblasts to root dentin conditioned with different endodontic irrigants: An experimental study

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          Abstract

          Background. Periradicular surgery is the last treatment option for teeth with persistent periradicular endodontic lesions. This study aimed to assess the adhesion of fibroblasts to root dentin conditioned with ethylenediaminetetraacetic acid (EDTA), MTAD, and QMix.

          Methods. Twelve dentin discs were fabricated of 6 human single-rooted teeth. Fibroblasts were isolated from the periodontal ligament (PDL) of a premolar tooth. The teeth were healthy and freshly extracted from the socket. The samples were divided into four groups for surface conditioning with (I) EDTA, (II) MTAD, (III) QMix, and the control group. Fibroblasts were cultured on conditioned dentin discs at 37°C, 95% air, and 5% CO 2 for 4 hours and then rinsed with PBS three times to eliminate unattached cells from the surface. The mean counts of attached cells were calculated using a Neubauer chamber. Also, the attachment of fibroblasts was evaluated using scanning electron microscopy (SEM).

          Results. The mean counts of fibroblasts attached to root dentin in EDTA, QMix, MTAD, and control groups were 303±46, 243±41, 213±33, and 347±38, respectively. No significant difference was noted in the number of fibroblasts attached between MTAD, EDTA, and QMix and the control group ( P>0.05). Under SEM, the fibroblasts were flat and spindle-shaped, with cytoplasmic processes covering the untreated dentin surface. In the experimental groups, the cells were rounder with fewer processes. All the three groups showed weaker adhesion to dentin compared to the control (untreated dentin) group.

          Conclusion. Under the limitations of this study, it was concluded that treating the dentin surface with EDTA, MTAD, or QMIX might not be an effective way to improve the adhesion of human PDL fibroblasts.

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          Most cited references33

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          Effect of chlorhexidine digluconate on different cell types: a molecular and ultrastructural investigation.

          Although several studies have shown that chlorhexidine digluconate (CHX) has bactericidal activity against periodontal pathogens and exerts toxic effects on periodontal tissues, few have been directed to evaluate the mechanisms underlying its adverse effects on these tissues. Therefore, the aim of the present study was to investigate the in vitro cytotoxicity of CHX on cells that could represent common targets for its action in the surgical procedures for the treatment of periodontitis and peri-implantitis and to elucidate its mechanisms of action. Osteoblastic, endothelial and fibroblastic cell lines were exposed to various concentrations of CHX for different times and assayed for cell viability and cell death. Also analysis of mitochondrial membrane potential, intracellular Ca2+ mobilization and reactive oxygen species (ROS) generation were done in parallel, to correlate CHX-induced cell damage with alterations in key parameters of cell homeostasis. CHX affected cell viability in a dose and time-dependent manners, particularly in osteoblasts. Its toxic effect consisted in the induction of apoptotic and autophagic/necrotic cell deaths and involved disturbance of mitochondrial function, intracellular Ca2+ increase and oxidative stress. These data suggest that CHX is highly cytotoxic in vitro and invite to a more cautioned use of the antiseptic in the oral surgical procedures.
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            Substantivity of chlorhexidine to human dentin.

            To better comprehend the role of CHX in the preservation of resin-dentin bonds, this study investigated the substantivity of CHX to human dentin. Dentin disks (n=45) were obtained from the mid-coronal portion of human third molars. One-third of dentin disks were kept mineralized (MD), while the other two-thirds had one of the surfaces partially demineralized with 37% phosphoric acid for 15 s (PDD) or they were totally demineralized with 10% phosphoric acid (TDD). Disks of hydroxyapatite (HA) were also prepared. Specimens were treated with: (1) 10 microL of distilled water (controls), (2) 10 microL of 0.2% chlorhexidine diacetate (0.2% CHX) or (3) 10 microL of 2% chlorhexidine diacetate (2% CHX). Then, they were incubated in 1 mL of PBS (pH 7.4, 37 degrees C). Substantivity was evaluated as a function of the CHX-applied dose after: 0.5 h, 1 h, 3 h, 6 h, 24 h, 168 h (1 week), 672 h (4 weeks) and 1344 h (8 weeks) of incubation. CHX concentration in eluates was spectrophotometrically analyzed at 260 nm. Significant amounts of CHX remained retained in dentin substrates (MD, PPD or TDD), independent on the CHX-applied dose or time of incubation (p<0.05). High amounts of retained CHX onto HA were observed only for specimens treated with the highest concentration of CHX (2%) (p<0.05). The outstanding substantivity of CHX to dentin and its reported effect on the inhibition of dentinal proteases may explain why CHX can prolong the durability of resin-dentin bonds. Copyright 2010 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.
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              Antibacterial and smear layer removal ability of a novel irrigant, QMiX.

              To assess in a laboratory experimental model the efficacy of a novel root canal irrigant, QMiX, against Enterococcus faecalis and mixed plaque bacteria in planktonic phase and biofilms. In addition, its ability to remove smear layer was examined. Enterococcus faecalis and mixed plaque bacteria were exposed to QMiX, 2% chlorhexidine (CHX), MTAD and 1% sodium hypochlorite (NaOCl) for 5 s, 30 s and 3 min. Following exposure, samples were taken, serially diluted and grown aerobically and anaerobically on tryptic soy agar (TSA) plates or on blood agar plates for 24 and 72 h, respectively, to measure killing of bacteria. E. faecalis and plaque biofilms were grown for 3 weeks on collagen-coated hydroxyapatite or dentine discs and exposed for 1 and 3 min to QMiX, 2% CHX, MTAD, 1% and 2% NaOCl. The amount of killed bacteria in biofilms was analysed by confocal laser scanning microscopy using viability staining. Dentine blocks were exposed to QMiX and 17% EDTA for 5 min. The effectiveness of smear layer removal by the solution was evaluated using scanning electron microscopy. For statistical analysis, one-way analysis of variance and comparison of two proportions were used. QMiX and 1% NaOCl killed all planktonic E. faecalis and plaque bacteria in 5 s, while 2% CHX and MTAD were unable to kill all plaque bacteria in 30 s, and some E. faecalis cells survived even 3 min of exposure. QMiX and 2% NaOCl killed up to 12 times more biofilm bacteria than 1% NaOCl (P < 0.01), 2% CHX (P < 0.05; P < 0.001) and MTAD (P < 0.05; P < 0.001). QMiX removed smear layer equally well as EDTA (P = 0.18 × 10(-5)). QMiX and NaOCl were superior to CHX and MTAD under laboratory conditions in killing E. faecalis and plaque bacteria in planktonic and biofilm culture. Ability to remove smear layer by QMiX was comparable to EDTA. © 2011 International Endodontic Journal.
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                Author and article information

                Journal
                J Dent Res Dent Clin Dent Prospects
                J Dent Res Dent Clin Dent Prospects
                J Dent Res Dent Clin Dent Prospects
                TBZMED
                Journal of Dental Research, Dental Clinics, Dental Prospects
                Tabriz University of Medical Sciences
                2008-210X
                2008-2118
                Winter 2022
                29 May 2022
                : 16
                : 1
                : 11-17
                Affiliations
                1Department of Endodontics, School of Dentistry, Hamadan University of Medical Sciences, Hamadan, Iran
                2Research Center for Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran
                3Department of Biostatistics, School of Public Health and Research Center for Health Sciences, Hamadan University of Medical Sciences, Hamadan, Iran
                Author notes
                [* ] Corresponding author: Elham Khoshbin, Email: maryamkhalili698@ 123456gmail.com
                Author information
                https://orcid.org/0000-0002-5068-8080
                https://orcid.org/0000-0002-0122-4650
                Article
                10.34172/joddd.2022.002
                9339743
                38fa0f5a-2b5f-4ad3-a113-a59b0a699b7d
                ©2022 The Author(s).

                This is an open-access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium provided the original work is properly cited.

                History
                : 22 June 2021
                : 07 January 2022
                Page count
                Figures: 4, Tables: 1, References: 34, Pages: 7
                Categories
                Original Article

                Dentistry
                cell adhesion,ethylenediaminetetraacetic acid,periradicular surgery,scanning electron microscopy

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