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1.
Artículo en Inglés | MEDLINE | ID: mdl-38976309

RESUMEN

Purpose: 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-α-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)-1ß, 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-1ß, 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.

2.
J Trace Elem Med Biol ; 84: 127466, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38692230

RESUMEN

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-1ß (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-1ß decreased cell viability that was overcome by all concentrations of BA at both 24 and 72 h. The IL-1ß addition to the media increased the expressions of the proinflammatory cytokines IL-1ß, IL-6, IL-8, and IL-17; the anti-inflammatory cytokine IL-10; and the oxidative stress enzymes superoxide dismutase (SOD0 and glutathione peroxidase (GPX). The IL-1ß induced increase mRNA expression of IL-1ß 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-1ß 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-1ß induced expression of IL-8 at both 24 and 72 h. The induced increase in IL-17 by IL-1ß was not significantly affected by the BA additions. The increase in the anti-inflammatory cytokine IL10 induced by IL-1ß 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-1ß 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.


Asunto(s)
Ácidos Bóricos , Supervivencia Celular , Fibroblastos , Encía , Interleucina-1beta , Humanos , Ácidos Bóricos/farmacología , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Encía/citología , Encía/efectos de los fármacos , Interleucina-1beta/metabolismo , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Inflamación/tratamiento farmacológico , Inflamación/metabolismo , Inflamación/inducido químicamente , Estrés Oxidativo/efectos de los fármacos , Citocinas/metabolismo , ARN Mensajero/metabolismo , ARN Mensajero/genética
3.
J Periodontal Res ; 59(1): 63-73, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38069670

RESUMEN

BACKGROUND/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, ß-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 ß-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.


Asunto(s)
Cemento Dental , beta Catenina , beta Catenina/metabolismo , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Ácido Hialurónico/farmacología , Línea Celular , Osteocalcina/metabolismo , Sialoproteína de Unión a Integrina/metabolismo , Diferenciación Celular , Movimiento Celular , ARN Mensajero/metabolismo
4.
J Periodontol ; 2023 Oct 03.
Artículo en Inglés | MEDLINE | ID: mdl-37787060

RESUMEN

BACKGROUND: 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 µL) 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.

5.
J Periodontol ; 94(11): 1351-1362, 2023 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-37322861

RESUMEN

BACKGROUND: Resolvins 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. METHODS: Immortalized 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-κB) (RANK), receptor activator of NF-κB 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-α}, interleukin {IL}-1ß, 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). RESULTS: Both 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). CONCLUSIONS: RvD1 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.


Asunto(s)
Cemento Dental , Ácidos Docosahexaenoicos , Ácido Eicosapentaenoico/análogos & derivados , Inhibidor Tisular de Metaloproteinasa-2 , Ratones , Animales , Cemento Dental/metabolismo , Inhibidor Tisular de Metaloproteinasa-2/metabolismo , Inhibidor Tisular de Metaloproteinasa-1/metabolismo , Metaloproteinasa 3 de la Matriz , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Sialoproteína de Unión a Integrina/metabolismo , ARN Mensajero/metabolismo
6.
Biol Trace Elem Res ; 201(3): 1174-1180, 2023 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-35441257

RESUMEN

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-1ß, 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-α, IL-1ß, 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.


Asunto(s)
Citocinas , Nicotina , Humanos , Citocinas/metabolismo , Nicotina/farmacología , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Fibroblastos , Células Cultivadas
7.
Int J Vitam Nutr Res ; 92(3-4): 231-239, 2022 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-32718217

RESUMEN

This study examined the effect of curcumin on T-helper (Th17) and T-regulatory (Treg) cells regarding the mRNA of cytokines/mediators in the gingiva. Thirty-five male albino Wistar rats were divided into four groups: Group 1: periodontitis (n = 9); Group 2: periodontitis with curcumin treatment (n = 8); Group 3: periodontally healthy with curcumin treatment (n = 10); and Group 4: periodontally healthy (n = 8). Curcumin was administered via oral gavage (30 mg/kg/day) for a total of 15 days. The gingival tissues were investigated regarding mRNA expressions of Th17/Treg cytokines with qRT-PCR. The distributional properties of the data were evaluated using the Anderson-Darling normality test. Kruskal-Wallis and Mann-Whitney U tests were employed for multiple group comparisons. Partial least squares regression discriminant analysis (PLS-DA) was used to evaluate the degree of contribution of each mRNA to the separation of treatment groups. When the periodontitis groups were compared, curcumin treatment resulted in lower IL-1ß (Group 2 median: 0.002, Group 1 median: 0.12) and IL-6 (Group 2 median: 0.031, Group 1 median: 0.078) and higher IL-17 (Group 2 median: 1.07, Group 1 median: 0.583) relative mRNA expression in Group 2 than in Group 1 (p < 0.001). Group 3 also had higher IL-10 relative expression (median: 0.067) than Groups 1 and 4 (median: 0.028, 0.007, respectively. p < 0.001). These results indicate that curcumin might be a promising agent for the prevention and/or treatment of periodontal diseases due to its decreasing effect on IL-1ß and IL-6 mRNA expression.


Asunto(s)
Curcumina , Periodontitis , Animales , Curcumina/farmacología , Citocinas , Interleucina-6/genética , Masculino , Periodontitis/tratamiento farmacológico , Periodontitis/metabolismo , ARN Mensajero/genética , Ratas
8.
Noncoding RNA Res ; 6(1): 15-22, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33385103

RESUMEN

MicroRNA (miRNA) expression is a dynamic process in the cell, and the proper time period for post-transcriptional regulation might be critical due to the gene-on/-off expression times of the cell. Here, we investigated the effect of different time-points on proliferation, invasion and miRNA expression profiles of human breast cancer cell lines MCF-7 (non-metastatic, epithelium-like breast cancer cell line with oestrogen receptor (ER) positive (+) and human breast cancer cell lines MDA-MB-435 (metastatic, invasive, ER negative (-). For this purpose, MCF-7 and MDA-MB-435 cells were seeded different number in E-plate 16 for proliferation experiment using an electrical impedance-based real-time cell analyzer system (RTCA) for 168 h. Similarly, invasion potential of MCF-7 and MDA-MB-435 were determined by RTCA for 90 h. Total RNAs including miRNAs were isolated at 2, 4, 6, 12, 24, 48 h from the MCF-7 and MDA-MB-435 cells. Afterward, the quantitative 84 miRNA expressions of MCF-7 and MDA-MB-435 were analyzed by Fluidigm Microfluidic 96.96 Dynamic Array. The results of these study demonstrated that both proliferation potential and invasion capacity of MDA-MB-435 is higher than MCF-7 as time-dependent manner. Furthermore, we detected that up/down expressions of 32 miRNAs at all time points in MDA-MB-435 compared to MCF-7 (at least ten-fold increased). Because of the high number of miRNAs, we more closely evaluated the expression of six of them (miR-100-5p, miR-29a-3p, miR-130a-3p, miR-10a-5p, miR-10b-5p, miR-203a), and determined that their levels were dramatically changed by at least 50-fold at different time points of the experiment (p < 0.01). The expression levels of five of these miRNAs (miR-100-5p, miR-10a-5p, miR-10b-5p, miR-130a-3p, and miR-29a-3p) started to increase from the fourth hour and continued to increase until the 48th hour in MDA-MB-435 cells compared to MCF-7 cells (p < 0.01). Simultaneously, the expression of one of these miRNAs (miR-203a) decreased from the sixth hour to the 48th hour in MDA-MB-435 as compared to MCF-7. We determined pathways associated with target genes using mirPath - DIANA TOOLS. Small RNAs including miRNA are essential regulatory molecules for gene expressions. In the literature, gene expressions have been published as burst and pulse in the form of discontinuous transcription. The data of the research suggested that time-dependent changes of miRNA expressions can be affected target gene transcriptional fluctuations in breast cancer cell and can be base for the further studies.

9.
Biol Trace Elem Res ; 199(4): 1534-1543, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-32594357

RESUMEN

The effects of boron on the formation and maintenance of mineralized structures at the molecular level are still not clearly defined. Thus, a study was conducted using MC3T3-E1 cells to determine whether boron affected mRNA expressions of genes associated with bone/alveolar bone formation around the teethMC3T3-E1 (clone 4) cells were cultured in media treated with boric acid at concentrations of 0, 0.1, 10, 100, or 1000 ng/ml. Total RNAs of each group were isolated on day 3. Gene expression profiles were determined by using RT2 Profiler PCR micro-array that included 84 genes associated with osteogenic differentiation. Tuftelin1 mRNA expression was upregulated by all boron treatments. The upregulation was confirmed by quantitative RT-PCR using the tuftelin probe. While 100 ng/ml had no effect on the integrin-α2 (Itga2) transcript and 1 ng/ml boric acid induced Itga2 mRNA expression (2.1-fold), 0.1, 10, and 1000 ng/ml boric acid downregulated the integrin-α2 gene transcript 2.2-, 1.5-, and 2.1-fold respectively. While 0.1 ng/ml boric acid induced BMP6, increased BMP1r mRNA expression (1.5 fold) was observed in 1000 ng/ml boric acid treatment. The findings suggest that boron affects the regulation of the tuftelin1 gene in osteoblastic cells. Further studies are needed to establish that the beneficial actions of boron on alveolar bone and tooth formation and maintenance include an effect on the expression of the tuftelin1 gene.


Asunto(s)
Boro , Osteogénesis , Ácidos Bóricos , Boro/farmacología , Diferenciación Celular , Proteínas del Esmalte Dental , Osteoblastos , ARN Mensajero/genética
10.
J Endod ; 46(7): 957-963, 2020 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-32439225

RESUMEN

INTRODUCTION: The aim of this study was to determine the effect of the continuous wave of condensation technique (CWCT) and the thermoplastic gutta-percha injection (TGI) technique on the messenger RNA (mRNA) expressions of heat shock proteins (HSPs) and mineralized tissue-associated proteins of the immortalized mouse cementoblasts (OCCM.30). METHODS: Crowns of human premolar teeth with single and straight canals were removed. The root canals were prepared up to the ProTaper Next X5 file (Dentsply Maillefer, Ballaigues, Switzerland) in combination with 2 mL 2.5% sodium hypochlorite solution. Roots (12 ± 2 mm height) were sterilized (121°C for 20 minutes) and placed vertically to the cell culture dishes using a tissue culture insert by opening holes according to the root diameter after the removal of 1 mm from the apex for appropriate adaptation to the petri dish surfaces. Six groups were created: control 1 (without teeth), control 2 (with teeth), AH Plus (Dentsply DeTrey, Konstanz, Germany), single-cone obturation (SC), CWCT, and thermoplastic gutta-percha injection technique (TGI). The viability of the OCCM.30 cells was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell viability experiments at 24 and 96 hours. The mRNA expression of HSP27, HSP70, and HSP90 and mineralized tissue markers (bone sialoprotein, osteocalcin, runt-related transcription factor 2, type I collagen, and alkaline phosphatase) was evaluated by real-time polymerase chain reaction. RESULTS: Reduced OCCM.30 cell viability was observed in all groups except the control groups. When the SC technique and CWCT and TGI groups were compared, it was observed that heat had a significant negative effect on cell viability (P < .05). A reduction in the mRNA expressions of HSP27, HSP70, and HSP90 was recognized in all test groups when compared with the control 1 group (P < .01). When the warm gutta-percha techniques were compared with the SC technique, a decrease in mRNA expression of HSP27 and HSP90 was noted (P < .01). The HSP70 transcript was similar in the CWCT group and the SC group. Higher HSP70 mRNA expression was observed in the TGI group compared with the SC group. In all groups except the control 1 group, bone sialoprotein, osteocalcin, runt-related transcription factor 2, type I collagen, and alkaline phosphatase mineralized tissue markers were affected, but this negative effect was higher in the heat-treated groups (P < .05). CONCLUSIONS: Within the limitations of this study, it was concluded that warm gutta-percha techniques reduced the mRNA expressions of the genes for HSPs and mineralized tissue-associated proteins of cementoblasts. Further animal studies are needed to clarify the effect of heat on the behavior of cementoblasts histologically in short- and long-term periods.


Asunto(s)
Gutapercha , Materiales de Obturación del Conducto Radicular , Animales , Supervivencia Celular , Cemento Dental , Cavidad Pulpar , Alemania , Respuesta al Choque Térmico , Humanos , Ratones , Obturación del Conducto Radicular , Preparación del Conducto Radicular
11.
J Periodontol ; 91(6): 800-808, 2020 06.
Artículo en Inglés | MEDLINE | ID: mdl-31489997

RESUMEN

BACKGROUND: Previous studies reported that nicotine, which is the prominent constituent of tobacco, has negative effects on periodontium cells. However, the precise role of nicotine in cementoblast functions remains unclear. In the present study, we investigated the effects of nicotine on the functions of cementoblasts (OCCM-30) in terms of proliferation, migration, and mineralized tissue-associated gene expression. METHODS: Immortalized murine cementoblasts were exposed to various concentrations (0, 10-6 , 10-5 , 10-4 , 10-3 , 10-2 , 10-1 , 1, 2.5, 5, and 10 mM) of nicotine, and cementoblast proliferation was then evaluated using a real-time cell analyzer for 142 hours. Using an in vitro wound healing assay, cell migration was evaluated 2, 4, 6, and 24 hours after exposure to different concentrations of nicotine (1, 2.5, 5, and 10 mM). The mRNA expressions of bone sialoprotein (BSP), collagen type I (COL-I), osteocalcin (OCN), runt-related transcription factor 2 (Runx2), and alkaline phosphatase (ALP) were assessed in the nicotine-treated (0, 10-3 , 10-2 , 10-1 , 1, 2.5, 5, and 10 mM) OCCM-30 cells by reverse transcription quantitative polymerase chain reaction at 8 and 24 hours exposure. RESULTS: At concentrations of 1 to 10 mM, nicotine significantly reduced cementoblast proliferation (P <0.01). Exposure to nicotine at other concentrations (1, 2.5, and 5 mM) significantly reduced wound healing rates, whereas nicotine at a concentration of 10 mM immediately decreased the viability of OCCM-30 cells. Similar results were observed in inverted microscopy images at the highest nicotine concentrations. All concentrations of nicotine decreased the transcripts of BSP and COL-I in a dose- and time-dependent manner (P <0.001). Nicotine concentrations higher than 1 mM reduced the expression of OCN, RunX2, and ALP in a time-dependent manner (P <0.001). CONCLUSIONS: This study indicated that nicotine inhibited the proliferation, migration, and mineralized tissue-associated gene expression of OCCM-30 cells. These findings suggest that nicotine negatively affects cementoblast function and the formation of new cementum, which is critical for new attachment.


Asunto(s)
Cemento Dental , Nicotina , Animales , Diferenciación Celular , Proliferación Celular , Cemento Dental/metabolismo , Sialoproteína de Unión a Integrina/genética , Ratones , Nicotina/farmacología , Osteocalcina/genética
12.
Lasers Med Sci ; 32(4): 911-919, 2017 May.
Artículo en Inglés | MEDLINE | ID: mdl-28332131

RESUMEN

The aim of this study was to evaluate the effects of diode laser biostimulation on cementoblasts (OCCM.30). A total of 40 root plates were obtained from healthy third molar teeth and assigned to the following two groups: (1) control group and (2) laser-treated group. Root plates were placed into the cell culture inserts, and OCCM.30 cells were seeded onto root plates. Cells were irradiated with a low level of diode laser (power: 0.3 W in continuous wave, 60 s/cm2). Proliferation and mineralized tissue-associated gene's and BMP's messenger RNA (mRNA) expressions of cementoblasts were evaluated. Total RNAs were isolated on day 3 and integrin-binding sialoprotein (Ibsp), bone gamma-carboxyglutamate protein (Bglap), Type I collagen (Col1a1), osteoblastic transcription factor, runt-related transcription factor (Runx2), and Bone Morphogenetic Protein (BMP)-2, 3, 4, 6, and 7 mRNA expressions were determined using quantitative RT-PCR. von Kossa staining was performed to evaluate biomineralization of OCCM.30 cells. In the proliferation experiment, while there was no significant difference until 96 h, laser irradiation retarded the decrease in cell proliferation trend after 96 h compared to the untreated control group. Statistically significant increase in Ibsp, Bglap, and BMP-2,3,6,7 mRNA expressions were noted in the laser groups when compared to the untreated control group (p < 0.05). Laser irradiation induced mineralized nodule formation of cementoblasts. The results of this study reveal that the biostimulation setting of diode laser modulates the behavior of cementoblasts inducing mineralized tissue-associated gene's mRNA expressions and mineralization. Therefore, biostimulation can be used during regenerative periodontal therapies to trigger cells with periodontal attachment apparatus.


Asunto(s)
Cemento Dental/efectos de la radiación , Láseres de Semiconductores , Terapia por Luz de Baja Intensidad , Animales , Calcificación Fisiológica/genética , Calcificación Fisiológica/efectos de la radiación , Adhesión Celular/efectos de la radiación , Línea Celular , Proliferación Celular/efectos de la radiación , Supervivencia Celular/efectos de la radiación , Regulación de la Expresión Génica/efectos de la radiación , Ratones , Diente Molar/efectos de la radiación , ARN Mensajero/genética , ARN Mensajero/metabolismo , Raíz del Diente/química , Raíz del Diente/efectos de la radiación
13.
J Oral Maxillofac Surg ; 73(8): 1562.e1-10, 2015 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-25930954

RESUMEN

PURPOSE: To evaluate the effects of bioresorbable fixation screws (BFSs) on human gingival fibroblast (HGF) and mouse osteoblast (MC3T3-E1) cell viability. MATERIALS AND METHODS: The KLS Martin SonicPins Rx, Synthes RapidSorb Cortex Screws, and Inion CPS Bioabsorbable Fixation System each were incubated in Dulbecco's Modified Eagle Medium for 72 hours according to ISO 10993-5 standards. A real-time cell analyzer was used to evaluate cell survival. After seeding 200-µL cell suspensions in the wells of an E-plate View 96, HGF and MC3T3-E1 cells were treated with the bioactive components released by the bioresorbable materials and monitored every 15 minutes for 96 hours. Statistical significance was determined using 1-way analysis of variance and Tukey-Kramer tests. RESULTS: There were significant differences in the HGF responses to the untreated control conditions and the Synthes (P < .01), Inion (P < .05), and KLS Martin (P < .05) treatments over 48 hours. The Synthes (P < .01) and Inion (P < .01) treatments produced lower HGF cell index values than the untreated control at 72 hours, whereas the KLS Martin treatment did not. When left to elute for 96 hours, there were no significant differences in values among the control and study groups for HGFs (P > .05). All tested BFSs decreased cell survival rates of M3T3C1 cells for 48 hours (P < .01), 72 hours (P < .001), and 96 hours (P < .001). CONCLUSION: Differences in the sensitivities of the 2 tested cell lines to the different BFSs might be the result of the different materials used to manufacture the screws. These results provide fundamental knowledge and new insights for the future design and development of new biocompatible BFSs for oral and maxillofacial surgery.


Asunto(s)
Materiales Biocompatibles , Tornillos Óseos , Encía/citología , Osteoblastos/citología , Células 3T3 , Animales , Supervivencia Celular , Fibroblastos/citología , Humanos , Ratones , Pruebas de Toxicidad
14.
Dent Mater J ; 34(2): 154-60, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-25736260

RESUMEN

To evaluate the cytotoxicity of resin cements on dental pulp-derived cells (bDPCs), Bifix QM (BQM), Choice 2(C2), RelyX U200(RU200), Maxcem Elite(ME), and Multilink Automix(MA) were tested. The materials were incubated in DMEM for 72 h. A real-time cell analyzer was used to evaluate cell survival. The statistical analyses used were one-way ANOVA and Tukey-Kramer tests. BQM, RU200, and ME demonstrated a significant decrease in the bDPCs' index at 24 and 72 h (p≤0.001). These materials were found to be the most toxic resin cements, as compared to the control and other tested materials (C2 and MA). However, C2 and MA showed a better survival rate, compared to BQM, RU200, and ME, and had lower cell index than the control group. The cytotoxic effects of resin cements on pulpa should be evaluated during the selection of proper cements.


Asunto(s)
Pulpa Dental/efectos de los fármacos , Cementos de Resina/efectos adversos , Animales , Bovinos , Células Cultivadas , Pulpa Dental/citología , Ensayo de Materiales , Cementos de Resina/farmacología
15.
J Adv Prosthodont ; 7(1): 21-6, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25722833

RESUMEN

PURPOSE: To evaluate the cytotoxicity of temporary luting cements on bovine dental pulp-derived cells (bDPCs). MATERIALS AND METHODS: Four different temporary cements were tested: Rely X Temp E (3M ESPE), Ultratemp (Ultradent), GC Fuji Temp (GC), and Rely X Temp NE (3M ESPE). The materials were prepared as discs and incubated in Dulbecco's modified eagle's culture medium (DMEM) for 72 hours according to ISO 10993-5. A real-time cell analyzer was used to determine cell vitality. After seeding 200 µL of the cell suspensions into the wells of a 96-well plate, the bDPCs were cured with bioactive components released by the test materials and observed every 15 minutes for 98 hours. One-way ANOVA and Tukey-Kramer tests were used to analyze the results of the proliferation experiments. RESULTS: All tested temporary cements showed significant decreases in the bDPCs index. Rely X Temp E, GC Fuji Temp, and Rely X Temp NE were severely toxic at both time points (24 and 72 hours) (P<.001). When the cells were exposed to media by Ultratemp, the cell viability was similar to that of the control at 24 hours (P>.05); however, the cell viability was significantly reduced at 72 hours (P<.001). Light and scanning electron microscopy examination confirmed these results. CONCLUSION: The cytotoxic effects of temporary cements on pulpal tissue should be evaluated when choosing cement for luting provisional restorations.

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