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Öğe Boric acid alleviates periodontal inflammation induced by IL-1? in human gingival fibroblasts(Elsevier Gmbh, 2024) Bozkurt, Serife Buket; Hakki, Sema S.; Nielsen, Forrest H.Background: Boric acid (BA) has been found to have therapeutic effects on periodontal disease through beneficially affecting antibacterial, anti-viral, and anti-inflammatory actions. Methods: This study was conducted to determine the effect of BA on cell viability and on mRNA expressions of proinflammatory and anti-inflammatory cytokines and on oxidative stress enzymes induced by IL-1 beta (1 ng/mL) in Human Gingival Fibroblasts (HGF) cultured for 24 and 72 h in DMEM media. The BA concentrations added to the media were 0.09 %, 0.18 %, 0.37 %, and 0.75 %. Results: All of the BA concentrations increased the viability of cell cultured in DMEM media only, indicating that these concentrations were not toxic and actually beneficial to cell viability. The addition of 1 ng/m: of IL-1 beta decreased cell viability that was overcome by all concentrations of BA at both 24 and 72 h. The IL-1 beta addition to the media increased the expressions of the proinflammatory cytokines IL-1 beta, IL-6, IL-8, and IL-17; the antiinflammatory cytokine IL-10; and the oxidative stress enzymes superoxide dismutase (SOD0 and glutathione peroxidase (GPX). The IL-1 beta induced increase mRNA expression of IL-1 beta was decreased at 24 h by the 0.37 % and 0.75 % BA additions to the media and decreased in a dose-dependent manner by all concentrations of BA at 72 h. The IL-1 beta induced increase in the expression of IL-6 was decreased in dose-dependent manner at 72 h by BA. All BA concentrations decreased the IL-1 beta induced expression of IL-8 at both 24 and 72 h. The induced increase in IL17 by IL-1 beta was not significantly affected by the BA additions. The increase in the anti-inflammatory cytokine IL10 induced by IL-1 beta was increased further by all BA additions in dose dependent manner at both 24 and 72 h. The mRNA expressions of SOD and GPX increased by IL-1 beta were further increased by the 0.37 % and 0.75 % BA concentrations at 72 h. Conclusions: These findings indicate that BA can significantly modulate the cytokines that are involved in inflammatory stress and reactive oxygen species action and thus could be an effective therapeutic agent in the treatment of periodontal disease.Öğe Boric Acid Reverses Nicotine-Induced Cytokine Expressions of Human Gingival Fibroblasts(Springernature, 2023) Bozkurt, Serife Buket; Nielsen, Forrest H.; Hakki, Sema S.Nicotine, the major bioactive ingredient in tobacco, is a major risk factor for periodontal disease and destruction. Nicotine has been shown to stimulate the production of cytokines that are priming agents for inflammation that induces tissue destruction, such as IL-1 beta, IL-6, and IL-8, by gingival keratinocytes and human gingival fibroblasts (HGF). Boron as boric acid has been found to decrease pro-inflammatory cytokines and increase anti-inflammatory cytokines in cells with inflammatory stress. Thus, a study was performed to determine whether boric acid reverses negative effects of nicotine on human gingival fibroblasts (HGFs). The viability and cytokine expressions of HGFs cultured for 24 and 72 h in control medium with no nicotine or boric acid added and in media containing only nicotine, only boric acid, or a combination of BA and nicotine were determined. Nicotine in concentrations of 10(-1), 10(-2), 10(-3),10(-4), 10(-5), and 10(-6) mM significantly reduced cell viability compared to the control. Boric acid at 10 and 50 ng/mL in the media partially restored and 100 ng/mL in the media fully restored the nicotine-depressed HGF cell viability to the same level as the control group. Nicotine elevated the expression of pro-inflammatory cytokines TNF-alpha, IL-1 beta, IL-6, IL-8, and IL-17 and decreased the anti-inflammatory IL-10 in HGFs at 24 and 72 h. Boric acid at 100 ng/mL in the medium prevented the changes induced by nicotine alone. The findings indicate that boric acid can inhibit or reverse nicotine-induced pathology in periodontal tissue and thus may help maintain oral and periodontal health in tobacco users.Öğe Coenzyme Q10 and Vitamin E Regulate the Bioactivity of Human Corneal Fibroblast Cells(Mary Ann Liebert, Inc, 2024) Zor, Kursad Ramazan; Yilmaz, Ugur; Bozkurt, Serife BuketPurpose: Corneal fibroblasts are involved in the wound healing of the cornea with proliferation, migration, and differentiation processes. Coenzyme Q10 (CoQ10) and vitamin E can enhance corneal wound healing when applied after a corneal lesion as an eye drop. Thus, this study was performed to determine the potential efficiency of a CoQ10 ophthalmical solution containing a CoQ10 and vitamin E D-alpha-tocopherol polyethylene glycol 1000 succinate (TPGS)-derived formulation in human corneal fibroblasts (HCFs) in vitro.Methods: Primary HCFs were obtained from cadaveric corneal tissue, and cell viability was determined using MTT assay at 24 and 72 h. Cell migration was evaluated using an in vitro wound healing assay, and mRNA expressions of collagen type I (COL-I), collagen type III (COL-III), lumican, hyaluronan, matrix metalloproteinase (MMP)-1, MMP-2, MMP-9, tissue inhibitors of MMP (TIMP)-1, TIMP-2, interleukin (IL)-1 beta, IL-6, IL-8, and IL-10 were assessed using reverse transcription polymerase chain reaction at 24 and 72 h.Results: At various concentrations of CoQ10 ophthalmical solution (CoQ10-os), cell viability and wound healing rates of HCFs increased compared with the control group. The expressions of COL-I, COL-III, lumican, and hyaluronan were increased by CoQ10-os, whereas those of MMP-1, MMP-2, MMP-9, TIMP-1, TIMP-2, and TIMP-3 were not affected by CoQ10-os at 24 and 72 h. In treating HCFs with a CoQ10-os medium, IL-1 beta, IL-6, and IL-8 decreased, whereas IL-10 was significantly increased in a time- and dose-dependent manner.Conclusions: The findings indicate that CoQ10 and vitamin E-TPGS are potent regulators of the bioactivity of HCFs, thus supporting their potential application as ophthalmical solutions in therapies aimed at the fast regeneration of damaged cornea tissues.Öğe Differential effects of resolvin D1 and resolvin E1 on cementoblast function(Wiley, 2023) Bozkurt, Serife Buket; Hakki, Sema Sezgin; Kantarci, AlpdoganBackgroundResolvins are endogenous mediators of the resolution of inflammation. They are derived from omega-3 polyunsaturated fatty acid precursors. Resolvin D1 (RvD1) and Resolvin E1 (RvE1) are the best-characterized members for actively promoting periodontal regeneration in experimental animal models. Here, we evaluated the efficacy of RvD1 and RvE1 on cementoblasts, the key cells involved in dental cementum regeneration and the attachment of the tooth to the alveolar bone. MethodsImmortalized mouse cementoblasts (OCCM-30) were treated with different concentrations (0.1-1000 ng/mL) of RvD1 and RvE1. Cell proliferation was measured using an electrical impedance-based real-time cell analyzer. Mineralization was evaluated with von Kossa staining. The mRNA expression of mineralized tissue-associated markers of bone sialoprotein (BSP), Type I collagen (COL I), osteocalcin (OCN), osteopontin (OPN), runt-related transcription factor 2 (RunX2), alkaline phosphatase (ALP), osteoprotegerin (OPG), receptor activator of nuclear factor kappa B (NF-kappa B) (RANK), receptor activator of NF-kappa B ligand (RANKL), and extracellular matrix-degrading enzymes [matrix metalloproteinase (MMP)-1, MMP-2, MMP-3, MMP-9, and their tissue inhibitors (TIMP-1, TIMP-2)], RvE1 receptor (ChemR23) and RvD1 receptor (ALX/PFR2), cytokines (tumor necrosis factor-alpha {TNF-alpha}, interleukin {IL}-1 beta, IL-6, IL-8, IL-10, IL-17), oxidative stress enzymes [superoxide dismutase (SOD), glutathione peroxidase (GPX), and cyclooxygenase-2 (Cox-2)] were analyzed using quantitative polymerase chain reaction (qPCR). ResultsBoth RvD1 and RvE1 (10-100 ng/mL) significantly increased the proliferation of cementoblasts and mineralized nodules at all concentrations (p < 0.05). RvE1 increased BSP, RunX2, and ALP compared with the RvD1 dose and time-dependently, while RvD1 and RvE1 differentially regulated COL-I. RvE1 increased OPG mRNA expression, whereas RANK-RANKL mRNA expression decreased by RvE1. MMP-2, MMP-3, MMP-9, TIMP-1, and TIMP-2 expressions were reduced by RvE1 compared with RvD1. Treatment of cementoblasts with RvD1 and RvE1 differentially affected cytokine and oxidative stress enzymes while significantly increasing their receptor expressions (ChemR23 and ALX/PFR2). ConclusionsRvD1 and RvE1 regulate proliferation, mineralization, and gene expression in cementoblasts using similar pathways while differentially affecting tissue degradation, suggesting a targeted therapeutic approach for cementum turnover during periodontal regeneration.Öğe Hyaluronic acid enhances cell migration, viability, and mineralized tissue-specific genes in cementoblasts(Wiley, 2024) Hakki, Sema S.; Bozkurt, Serife Buket; Sculean, Anton; Bozic, DarkoBackground/Objectives: It has been repeatedly demonstrated that cementum formation is a crucial step in periodontal regeneration. Hyaluronic acid (HA) is an important component of the extracellular matrix which regulates cells functions and cell-cell communication. Hyaluronic acid/derivatives have been used in regenerative periodontal therapy, but the cellular effects of HA are still unknown. To investigate the effects of HA on cementoblast functions, cell viability, migration, mineralization, differentiation, and mineralized tissue-associated genes and cementoblast-specific markers of the cementoblasts were tested.Materials and Methods: Cementoblasts (OCCM-30) were treated with various dilutions (0, 1:2, 1:4, 1:8, 1:16, 1:32, 1:64, 1:128) of HA and examined for cell viability, migration, mineralization, and gene expressions. The mRNA expressions of osteocalcin (OCN), runt-related transcription factor 2 (Runx2), bone sialoprotein (BSP), collagen type I (COL-I), alkaline phosphatase (ALP), cementum protein-1 (CEMP-1), cementum attachment protein (CAP), and small mothers against decapentaplegic (Smad) -1, 2, 3, 6, 7, beta-catenin (Ctnnb1) were performed with real-time polymerase chain reaction (RT-PCR). Total RNA was isolated on days 3 and 8, and cell viability was determined using MTT assay on days 1 and 3. The cell mineralization was evaluated by von Kossa staining on day 8. Cell migration was assessed 2, 4, 6, and 24 hours following exposure to HA dilutions using an in vitro wound healing assay (0, 1:2, 1:4, 1:8).Results: At dilution of 1:2 to 1:128, HA importantly increased cell viability (p < .01). HA at a dilution of 1/2 increased wound healing rates after 4 h compared to the other dilutions and the untreated control group. Increased numbers of mineralized nodules were determined at dilutions of 1:2, 1:4, and 1:8 compared with control group. mRNA expressions of mineralized tissue marker including COL-I, BSP, RunX2, ALP, and OCN significantly improved by HA treatments compared with control group both on 3 days and on 8 days (p < .01). Smad 2, Smad 3, Smad 7, and beta-catenin (Ctnnb1) mRNAs were up-regulated, while Smad1 and Smad 6 were not affected by HA administration. Additionally, HA at dilutions of 1:2, 1:4, and 1:8 remarkably enhanced CEMP-1 and CAP expressions in a dilution- and time-dependent manner (p < .01).Conclusions: The present results have demonstrated that HA affected the expression of both mineralized tissue markers and cementoblast-specific genes. Positive effects of HA on the cementoblast functions demonstrated that HA application may play a key role in cementum regeneration.Öğe Local application of gingiva-derived mesenchymal stem cells on experimental periodontitis in rats(Wiley, 2024) Balaban, Yunus Emre; Akbaba, Sema; Bozkurt, Serife Buket; Buyuksungur, Arda; Akgun, E. Ece; Gonen, Zeynep Burcin; Salkin, HasanBackground: Stem cell-based approaches in regenerative periodontal therapy have been used in different experimental models. In this study, the effect of local application of gingival mesenchymal stem cells (GMSC) in fibroin/chitosan oligosaccharide lactate hydrogel (F/COS) on periodontal regeneration was evaluated using experimental periodontitis model in rats.Methods: Mesenchymal stem cells were isolated from the gingiva of rats and characterized. Viability tests and confocal imaging of GMSC in hydrogels were performed. Healthy control without periodontitis (Health; H; n=10), control with periodontitis but no application (Periodontitis; P; n=10), only hydrogel application (F/COS; n=10), and GMSC+F/COS (n=10) four groups were formed for in vivo studies. Experimental periodontitis was created with silk sutures around the maxillary second molars. GMSC labeled with green fluorescent protein (GFP) (250,000 cells/50 mu L) in F/COS were applied to the defect. Animals were sacrificed at 2nd and 8th weeks and maxillae of the animals were evaluated by micro-computed tomography (micro-CT) and histologically. The presence of GFP-labeled GMSC was confirmed at the end of 8 weeks.Results: Micro-CT analysis showed statistically significant new bone formation in the F/COS+GMSC treated group compared with the P group at the end of 8 weeks (p < 0.05). New bone formation was also observed in the F/COS group, but the statistical analysis revealed that this difference was not significant when compared with the P group (p > 0.05). Long junctional epithelium formation was less in the F/COS+GMSC group compared with the P group. Periodontal ligament and connective tissue were well-organized in F/COS+GMSC group.Conclusion: The results showed that local GMSC application in hydrogel contributed to the formation of new periodontal ligament and alveolar bone in rats with experimental periodontitis. Since gingiva is easly accessible tissue, it is promising for autologous cell-based treatments in clinical applications.Öğe RESOLVIN D1 (RvD1) REGULATES PORPHYROMONAS GINGIVALIS LIPOPOLYSACCHARIDE-INDUCED Del-1 AND CYTOKINE EXPRESSIONS IN HUMAN GINGIVAL FIBROBLASTS(Dokuz Eylul Univ Inst Health Sciences, 2023) Bozkurt, Serife Buket; Hakki, Sema S.Purpose: To detect the effect of Resolvin D1 (RvD1) on Developmental endothelial locus-1 (Del-1) and cytokine expressions of human gingival fibroblast cells exposed to Porphyromonas gingivalis lipopolysaccharide (P. gingivalis-LPS).Material and Methods: The effect of RvD1 on cell viability of human gingival fibroblasts exposed to P. gingivalis-LPS was determined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Meanwhile, the effect of RvD1 on Del-1 and cytokine (IL-1 & beta;, IL-6, IL-8, IL-10, IL-17) expressions of human gingival fibroblasts exposed to P. gingivalis-LPS (1000 ng/mL) were studied by real-time PCR experiment, statistical analysis was performed using GraphPad Prism version 5 for Windows.Results: Cell viability assay results demonstrated that RvD1 concentrations upregulated cell number compared to control group at 24 and at 72 h. While RvD1 reduced the IL-6, IL-8, and IL-17 mRNA expressions, the IL-10 and Del-1 mRNA expressions increased in a time-and dose-dependent manner. Also, IL-1 & beta; was not affected by RvD1 treatments.Conclusion: The increased expression of Del-1 and IL-10 by RvD1 down-regulated the pro-inflammatory cytokine expressions induced by P. gingivalis-LPS in gingival fibroblast. Resolvin D1displayed regulatory effects on gingival inflammation in P. gingivalis LPS-induced cell culture experiment. In particular, results of this study show that Del-1 induced by RvD1 may have therapeutic potential to modulate periodontal inflammation.Öğe The effect of boric acid on copine-7 expression and bioactivity in dental pulp stem cells(Turkish Energy Nuclear and Mining Research Institute, 2022) Bozkurt, Serife Buket; Hakki, Sema S.Boron has significant impact on the mineral composition of teeth. Copine-7 (Cpne-7) is secreted by pre-ameloblasts and induces dentin formation via differentiation of mesenchymal cells of dental. The goal of this study was to see how boric acid affected the bioactivity and expression of Cpne-7, collagen type I in dental pulp stem cell. The expression of Cpne-7, COL-I were assessed in the boric acid treated pulp cells by molecular method on 3 and 8 days exposure. When comparing the different boric acid concentrations to the control group in a proliferation experiment, no significant differences were noted. At 10 ng/mL boric acid, a rise in the number of mineralized nodules was observed. Boric acid concentrations (1 ng/mL, 10 ng/mL) increased the transcripts of Cpne-7 on day 3 and 8. Additionally, 1 ng/ mL boric acid concentration significantly upregulated of Cpne-7 expression compared to control group on day 3 and 8. When the 1-10 ng boric acid was compared to control group COL-I expression remarkably enhanced in cell. According to the current findings that boric acid may be a potent regulator of Cpne-7, which is a promising candidate for new dentin formation in regenerative dentistry, in dental pulp stem cells and provide osteogenic efficacy in therapies aimed at dentin formation. © 2022, Turkish Energy Nuclear and Mining Research Institute. All rights reserved.
