Human Gingival Fibroblast Cells Research Articles

Cytotoxicity of two different intercanal medicaments on human gingival fibroblasts - A Laboratory study

Aim: Intracanal medicaments are often recommended during endodontic sessions to eliminate the necrotic debris and microorganisms. The aim of the present study was to observe the cytotoxicity of calcium hydroxide (CH) and colchicine (COL), at different concentrations, on human gingival fibroblast cells.Materials and Methods: Human gingival fibroblasts were cultured on plastic flasks containing RPMI 1640 media and fetal calf serum 10% supplemented with antibiotic agents. Trypsin/ethylenediaminetetraacetic acid 0.2% enzyme was used to isolate the cells, and the suspension was transferred to tubes for centrifuging. Conventional CH and COL were separately mixed with sterile saline solution to prepare a stock media. By serial dilution of stock media, desired concentrations were prepared at 2, 1.75, 1.5, and 1.25 mg/ml, separately. After considering a control group, the cells were exposed to test materials. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was conducted at 24 h, 72 h, and 7 days later. Optical density (OD) was evaluated to attain cell viability percentage. Finally, the recorded data were analyzed by Kruskal–Wallis and Mann–Whitney tests using SPSS software version 19 at a significant level of 0.05.Results: The highest (1.40 ± 0.66) and lowest (0.15 ± 0.00) ODs were observed in CH 1.25 mg/ml and COL 1.5 mg/ml after 72 h, respectively. All of the concentrations of both CH and COL showed significant OD differences with the control group (all P = 0.001).Conclusion: Both CH and COL manifested similar cytotoxicity on human gingival fibroblast cells.

Phytochemical compound and non-cytotoxicity effect of sting bee and stingless bee honey against normal human gingival cell lines

Objective: Both honeybees (Apis spp.) and stingless bees (Trigona spp.) produce honeys which normally taken orally, have high nutritional and therapeutics value. Until recently, phytochemical comparison of both honey is still scarce and elucidating cytotoxicity effects on human gingival fibroblast cells (HGF) in oral cavity is of interest. Materials and Methods: Kelulut honey (KH), acquired from the stingless bees and acacia honey (AH) from the sting bees honey samples were underwent GC-MS analysis to ascertain their composition. HGF were exposed to various concentrations of KH and AH from the lowest 0.015% to the highest 5% by MTT assay for 24h, 48h and 72h. Results: GC-MS analysis determined various beneficial compounds such as flavonoids, furans, pyrans, levoglucosan and hydroxymethylfurfural from both of honey samples. MTT assay showed that the HGF cells demonstrated good viability up to percentages (v/v) as high as almost 2% in both honeys. The IC50 values for both honey for all time frames fall at above 2%. Conclusion: Both honey showed good survivability of HGF cells up to 2% of concentration. Bangladesh Journal of Medical Science Vol. 21(1) 2022 Page : 158-164

Cytotoxicity evaluation of different clear aligner materials using MTT analysis

Abstract Backround: The in vitro cytotoxic effects of six different clear aligner materials were evaluated using the MTT analysis. Methods: The clear aligner material samples [Duran (ScheuDental GmbH, Iserlohn, Germany), Zendura-Flx (Bay Materials LLC, Fremont, CA, USA), Taglus (Laxmi Dental Export Pvt. Ltd, Mumbai, India), Smart Track (Align Technology, San Jose, CA, USA), Zendura (Bay Materials LLC, Fremont, CA, USA), Essix C + (Essix® (Raintree Essix, Inc., 4001 Division St, Metairie, LA-USA)] were initially kept in a saline solution in airtight test tubes for 8 weeks at 37°C. According to the recommended ISO standards, the weights of the samples were divided by the volumes of the dilutions in the ratio of 0.1 g/ml. To evaluate the cytotoxicity of the samples, an MTT analysis was performed using a human gingival fibroblast cell line (HGF). To analyse the data, the Kruskal– Wallis test was applied (a=0.05). Results: Zendura was the most cytotoxic material resulting in 67.3 ± 16.20% cell viability, followed by Smart Track with 87.6 ± 5.53% cell viability. While Duran, Essix C + had 92.6 ± 26.34% and 94.9 ± 8.54% cell viability, Zendura-Flx, Taglus had 106.9 ± 12.76% and 113.183 ± 7.45% cell viability, respectively. Conclusion: While Zendura and Smart Track showed mild cytotoxicity, other materials showed greater cell viabilities. According to the ISO standards, the clinical use of each brand of aligners, except Zendura, may be considered reliable. Taking into account standard deviation, Zendura and Duran should be used with caution. The suppliers of aligners should adhere to the manufacturer’s recommendations since an increase in ion release might arise from material wear.

Antimicrobial and Anti-inflammatory Effects of α-Mangostin Soluble Film.

ABSTRACTObjectives:Plant-derived compounds are a major source of medicinal agents. Common oral diseases, including dental caries, periodontal disease, and candidiasis, are caused by biofilms. The nature of biofilm formations is complex, emphasizing the importance of finding novel products that possess bioactivity against microbes associated with those oral infections. The aims of this study were to determine the antimicrobial activity and antibiofilm formation of α-mangostin (α-MG) soluble film.Materials and Methods:Antimicrobial assays against Streptococcus mutans, Porphyromonas gingivalis, and Candida albicans were performed by identifying the minimal growth inhibition concentration and the minimal bactericidal concentration. Time-killing kinetic studies against the organisms and inhibition of biofilm formation were determined by the broth microdilution method. Human gingival fibroblast cell line and macrophage RAW267.4 cells were cultured, and the cell viability was assessed by the MTT assay. The anti-inflammatory effect of the α-MG film was investigated by measuring the inhibition of nitric oxide production.Results:The α-MG film demonstrated antimicrobial activity against the oral pathogens tested. The formulation reduced microbial growth about 1–3 Log CFU/mL at 2–4 h and complete killing at 24 h. No significant difference in inhibiting the biofilm formation of those three microorganisms was noted. In addition, the film containing α-MG demonstrated anti-inflammatory activity through the inhibition of nitric oxide production in a dose-dependent manner. The formulation was safe and showed no cytotoxicity at therapeutic dose.Conclusions:The α-MG film is effective against S. mutans, P. gingivalis, and C. albicans without significant cytotoxicity in vitro. Thus, this new product may have potential advantage in preventing those common oral infections.

Effect of nano zinc oxide on proliferation and toxicity of human gingival cells.

Periodontal dressing is used to cover the gum surface and protect the wound after periodontal surgery. Nanomaterials have been widely applied in dentistry in recent years. Zinc oxide (ZnO) is one of the main components of periodontal dressing. This study aims to explore the toxicity ZnO nanoparticles (ZnO NPs) causes to human gingival fibroblast cells (HGF-1) and its effect on cell proliferation. First, we identified and analyzed HGF-1, including cell morphology, growth curve, and immunohistochemistry staining. Then, we treated HGF-1 with ZnO NP. Cell viability, the integrity of the cell membrane, oxidative damage, and apoptosis were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) release assay, fluorescent probe, and flow cytometry. Furthermore, the expression of murine double minute 2 (MDM2) and p53 was determined by quantitative real-time polymerase chain reaction (qPCR) and Western blotting. We finally overexpressed MDM2 in HGF-1 to verify the relationship between MDM2 and cell proliferation. Our research indicated ZnO NPs did not affect cell proliferation at low concentrations. However, high-concentration ZnO NP inhibited cell proliferation, destroyed the integrity of cell membranes, and induced oxidative stress and apoptosis. In addition, high concentration of ZnO NPs inhibited the proliferation of HGF-1 by regulating the expression of MDM2 and p53. High concentration of ZnO NP caused toxicity to HGF-1 cells and inhibited cell proliferation by regulating MDM2 and p53 expression.

The Application of a Recombinant Antimicrobial Peptide of Thrombocidin-1 Expressed in Pichia pastoris as a Novel Approach Against Some Oral Pathogenic Bacteria: An In Vitro Study.

Oral infections and dental caries are considered serious health problems. Therefore, searching for new agents with antimicrobial properties seems to be crucial. This study aimed to evaluate the antimicrobial activity of the recombinant Thrombocidin-1 [TC-1] peptide on some oral pathogens. Also, the cytotoxicity of this peptide on human gingival fibroblast cells was investigated. In this study, Pichia pastoris was used for the expression of recombinant TC-1. The microbroth dilution method was used to determine the minimum inhibitory concentration [MIC] and minimum bacterial concentration [MBC]. It tested against four main oral pathogens; Streptococcus mutans, Streptococcus salivarius, Streptococcus oralis, and Enterococcus faecalis. Moreover, the cytotoxicity analysis was done on gingival fibroblast cells by the MTT method. The data were analyzed using a two-way analysis of variance [ANOVA] and Tukey's HSD tests. The most bactericidal effect of TC-1 was against S. salivarius, the highest bacteriostatic effect was against S. salivarius, and S. oralis had the lowest MIC value of 1.512 μg/ml. The Thrombocidin- 1 peptide showed lower antibacterial properties against E. faecalis compared with CHX, unlike the stronger antimicrobial effect on examined streptococci. According to cytotoxicity examination, no concentration of TC-1 presented over 50% growth inhibition [IC50] of the fibroblasts cells. Based on antimicrobial tests and cytotoxicity results, the Thrombocidin-1 peptide may be useful as a safe antibacterial agent against some oral pathogens in dental materials.

In Vitro Antimicrobial and Cytotoxicity Activities of Some Medicinal Plant Extracts against Oral Microbial Pathogens.

Medicinal plants have long been of great interest to scientists in the search for the best treatment of diseases, especially the infectious diseases. In recent years, the use of herbal medicines has become more well-known because of their antimicrobial, antifungal, anti-cancer and less side effects. The aim of this study was to investigate the antimicrobial and antifungal effects of Urtica dioica, Equisetum arvense, and Punica Granatum peel extracts on two common oral microorganisms, Streptococcus mutans and Candida albicans. The study investigated the hydro-alcoholic extract of the plants. The antimicrobial activity of the extracts was evaluated using the method of measuring the inhibition of microorganisms, and the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were determined using different concentrations of the extracts and also biofilm assay and SEM were determined. Also cell viability was assessed by MTT assay on human gingival fibroblast cells. The lowest MIC against S. mutants and C. albicans was related to the hydro-alcoholic extract of U. dioica. There was a significant reduction in the microbial biofilms by all three extracts. Among them, U. dioica could decrease the biofilms of S. mutans and C. albicans more than other extracts. In addition, the best results for growth inhibition zone were the hydro-alcoholic extracts of E. arvense and U. dioica with 35 and 30 mm growth zone, respectively. The results of SEM showed that P. granatum peel, U. dioica and E. arvense could destroy microbial biofilms without exerting any cytotoxic effects on HGF cell. The results of the study suggest that U. dioica, E. arvense, and P. Granatum peel extracts can be used as mouthwash with the least significant difference with routine mouthwashes. Also, the plant-based mouthwashes may be more suitable substitutes for chemical types in the future.

Effects of streptococcus mutans on the expression of inflammatory cytokine TNF-α and p53 by human gum fibroblast cells

Oral bacteria are involved in the etiology of oral cancer. In this study, Streptococcus mutans were examined for the gene’s expression involved in inflammation, carcinogenicity, and tumor inhibition among these species. This study aimed to determine the association between microbiome and oral cancer. Human Gingival Fibroblast (HGF) and S. mutans were cultured on their relative mediums. Then, HGF was preconditioned whit several concentrations of S. mutans extract, and the cell viability was determined by MTT assay. HGF cells were treated with an optimum dose of bacterial extract, and then the total RNA was extracted from treated and untreated HGF cells. Lastly, p53 and TNF-α gene expression was evaluated by Real-Time PCR. The results implied that in the S. mutans presence, TNF-α and P53 genes expression in the HGFs increased and decreased, respectively. This result suggests that S. mutans may invasively enhance oral cancer cells. These findings more designate the association between oral microbiota and oral cancer. This relation is of clinical importance for the prevention and treatment of oral cancer in the future. Keywords: streptococcus mutans; human gingival fibroblast (HGF); oral cancer; P53; TNF-α.

Comparison of physical and biological properties of a flowable fiber reinforced and bulk filling composites

ObjectiveTo evaluate in vitro the mechanical, biological, and polymerization behavior of a flowable bulk-fill composite with fibers as a dispersed phase. MethodsEverX Flow™ (GC Corporation) (EXF), one conventional bulk-fill composite (Filtek™ Bulk Fill Posterior Restorative, 3 M (FBF)), and one flowable bulk composite without fibers (SDR® flow+, Dentsply (SDR)) were tested. Samples were characterized in terms of flexural strength (ISO 4049), fracture toughness (ISO 20795-1), and Vickers hardness. Polymerization stress and volumetric shrinkage were evaluated. The in vitro biological assessment was achieved on cultured primary Human Gingival Fibroblast cells (HGF). The cell metabolic activity was evaluated using Alamar Blue assay at 1, 3, and 5 days of contact to the 3 tested composite extracts (ISO 10993) and cell morphology was evaluated by confocal microscopy. Data were submitted to One-Way analysis of variance (ANOVA) and independent t-test (α = 0.05). ResultsFBF showed statistically higher Vickers hardness and flexural modulus than EXF and SDR. However, EXF showed statistically higher KIC than FBF and SDR. EXF had the statistically highest shrinkage stress values and FBF the lowest. Archimedes volumetric shrinkage showed significantly lower values for FBF as compared to the other two composites. Slight cytotoxic effect was observed for the three composites at day one. An enhancement of metabolic activity at day 5 was observed in cells treated with EXF extracts. SignificanceEXF had a significantly higher fracture toughness validating its potential use as a restorative material in stress bearing areas. EXF showed higher shrinkage stress values, and less cytotoxic effect.Fiber reinforced flowable composite is mainly indicated for deep and large cavities, signifying the importance for assessing its shrinkage stress and biological behavior.

Influence of cold atmospheric plasma on dental implant materials \u2014 an in vitro analysis

Background and objectivesAlterations in the microenvironment of implant surfaces could influence the cellular crosstalk and adhesion patterns of dental implant materials. Cold plasma has been described to have an influence on cells, tissues, and biomaterials. Hence, the mechanisms of osseointegration may be altered by non-thermal plasma treatment depending on different chemical compositions and surface coatings of the biomaterial. The aim of the present study is to investigate the influence of cold atmospheric plasma (CAP) treatment on implant surfaces and its biological and physicochemical side effects.Materials and methodsDental implant discs from titanium and zirconia with different surface modifications were treated with CAP at various durations. Cell behavior and adhesion patterns of human gingival fibroblast (HGF-1) and osteoblast-like cells (MG-63) were examined using scanning electron microscopy and fluorescence microscopy. Surface chemical characterization was analyzed using energy-dispersive X-ray spectroscopy (EDS). Quantitative analysis of cell adhesion, proliferation, and extracellular matrix formation was conducted including real-time PCR.ResultsCAP did not affect the elemental composition of different dental implant materials. Additionally, markers for cell proliferation, extracellular matrix formation, and cell adhesion were differently regulated depending on the application time of CAP treatment in MG-63 cells and gingival fibroblasts.ConclusionsCAP application is beneficial for dental implant materials to allow for faster proliferation and adhesion of cells from the surrounding tissue on both titanium and zirconia implant surfaces with different surface properties.Clinical relevanceThe healing capacity provided through CAP treatment could enhance osseointegration of dental implants and has the potential to serve as an effective treatment option in periimplantitis therapy.

Effect of nano zinc oxide on proliferation and toxicity of human gingival cells.

Periodontal dressing is used to cover the gum surface and protect the wound after periodontal surgery. Nanomaterials have been widely applied in dentistry in recent years. Zinc oxide (ZnO) is one of the main components of periodontal dressing. This study aims to explore the toxicity ZnO nanoparticles (ZnO NPs) causes to human gingival fibroblast cells (HGF-1) and its effect on cell proliferation. First, we identified and analyzed HGF-1, including cell morphology, growth curve, and immunohistochemistry staining. Then, we treated HGF-1 with ZnO NP. Cell viability, the integrity of the cell membrane, oxidative damage, and apoptosis were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) release assay, fluorescent probe, and flow cytometry. Furthermore, the expression of murine double minute 2 (MDM2) and p53 was determined by quantitative real-time polymerase chain reaction (qPCR) and Western blotting. We finally overexpressed MDM2 in HGF-1 to verify the relationship between MDM2 and cell proliferation. Our research indicated ZnO NPs did not affect cell proliferation at low concentrations. However, high-concentration ZnO NP inhibited cell proliferation, destroyed the integrity of cell membranes, and induced oxidative stress and apoptosis. In addition, high concentration of ZnO NPs inhibited the proliferation of HGF-1 by regulating the expression of MDM2 and p53. High concentration of ZnO NP caused toxicity to HGF-1 cells and inhibited cell proliferation by regulating MDM2 and p53 expression.

In Vitro Biocompatibility of CPP-ACP and Fluoride-containing Desensitizers on Human Gingival Cells.

To analyze the biocompatibility of different desensitizers containing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) and fluoride in their composition: MI Varnish (MV), Clinpro White Varnish (3M Oral Care), Profluorid Varnish (VOCO), Duraphat (Colgate) and Embrace Varnish (Pulpdent) on human gingival fibroblast cells (hGF). Human gingival fibroblast (hGF) cells were exposed to several desensitizer extracts at different concentrations (0.1%, 1%, and 4% eluates). Then, in vitro biocompatibility was studied by analyzing the IC50 value, cell proliferation (MTT assay and cell cycle), cell migration (wound healing assay), cell morphology and F-actin content (immunocytofluorescence), and induction of apoptosis/necrosis (flow cytometry). Data were analyzed by one-way analysis of variance (ANOVA) followed by Tukey test. The lowest cell viability and IC50 were observed in all concentrations of Embrace Varnish-treated hGFs (p<0.001), whereas the highest were exhibited by those treated with Clinpro White Varnish. Similar effects were evidenced when induction of apoptosis/necrosis and cell migration assays were assessed. Finally, MI Varnish, Profluorid Varnish, Duraphat, and Embrace Varnish extracts showed lower numbers of attached cells, some of them with an unusual fibroblastic morphology when cultured with 4% concentration of the varnishes, while Clinpro White Varnish exhibited a similar number of cells with an evident actin cytoskeleton compared to the control group. The results obtained in this study indicate that hGFs show better in vitro biocompatibility after exposure to Clinpro White Varnish, even at the highest concentration employed, making it the most eligible for topical applications. In contrast, Embrace Varnish exhibited a high cytotoxicity towards hGFs that could potentially delay the healing process and regeneration of the oral mucosa, although more studies are needed to confirm this hypothesis.

A New Approach Against Some Oral Pathogenic Bacteria Using a Chimeric Antimicrobial Peptide Derived from the Camel Milk; Lactoferrampin - Lactoferricin Chimer.

The present study was conducted to evaluate the antimicrobial effects of the recombinant chimer present in the lactoferrampin-lactoferricin [LFA-LFC] derived from the camel milk on some oral bacteria responsible for dental caries and endodontic failures. The antimicrobial activity was assessed on the Streptococcus mutans [ATCC 35668], Streptococcus salivarius [ATCC 9222], Streptococcus oralis [ATCC 35037], and Enterococcus faecalis [ATCC 29212], using the microbroth dilution method. The cytotoxicity analysis was done through the MTT method on the human gingival fibroblasts. The data were reported using the descriptive methods and were analyzed by the one-way analysis of variance (ANOVA) and Tukey HSD test. Results showed that the chimeric peptide had the highest bacteriostatic effect on S. salivarius with the lowest minimum inhibitory concentration value of 1.22 μg/Ml. Also, LFA-LFC chimer was more effective against S. mutans and S. salivarius compared to using 0.2% chlorhexidine mouthwash. The minimum bactericidal concentration analysis showed the most bactericidal effect against S. mutans [1.256 μg/mL]. In spite of the greater antibacterial effect on the evaluated streptococci, this peptide showed lower bacteriostatic and bactericidal properties against E. faecalis compared to the chlorhexidine. Based on cytotoxicity assay, over 50% of the cells were viable in all the evaluation times, demonstrating the biocompatibility of the peptide. The LFA-LFC chimer revealed comparable or even more effective antibacterial properties compared to the chlorhexidine mouthwash against the caries-inducing bacteria with no toxicity on the human gingival fibroblast cells. So, this peptide can be used as a safe alternative to chlorhexidine and other chemicals in dental applications for the prevention and management of dental caries.

Mucoadhesive film containing \u03b1-mangostin shows potential role in oral cancer treatment

BackgroundOral cancer is often preceded by a mucosal lesion called an oral potentially malignant disorder (OPMD). Many plant-derived compounds are of value in medicine. The objectives of this study were to develop a soluble mucoadhesive film containing α-mangostin (α-MG), a compound extracted from the peel of mangosteen fruit, and determine its activities against oral cancer cells, against human papillomavirus type 16 (HPV-16) pseudovirus, and its anti-inflammatory properties.MethodsA soluble mucoadhesive film containing α-MG was prepared. Oral squamous carcinoma cell line (SCC25), murine macrophage cells (RAW264.7), and human gingival fibroblast cell line were cultured. Anticancer activity and viability of SCC25 cells in response to α-MG film solution were determined by MTT assay. HPV-16 pseudovirus was constructed and effects of the film solution on attachment and post-attachment steps of the infection were investigated. Anti-inflammatory activity was assessed by nitric oxide (NO) inhibition. Fibroblast cell migration was determined by in vitro scratch assay.ResultsThe soluble α-MG film showed cytotoxic effects on SCC25 cells in concentration > 125 µg/ml with IC50 of 152.5 µg/ml. Antiviral activity against HPV-16 pseudovirus was observed at attachment step, but not at post-attachment step. The film also possessed a strong anti-inflammatory effect and promoted wound healing without cytotoxicity.ConclusionsMucoadhesive film containing α-MG has a cytotoxic effect on oral squamous carcinoma cell line and an inhibitory effect on HPV-16 pseudovirus at attachment step. The α-MG film also shows a potent anti-inflammatory activity and enhances wound healing. Thus, the soluble α-MG film may have a potential role in treating oral cancer.

Fibroblast behavior on conventionally processed, milled, and printed occlusal device materials with different surface treatments

Statement of problemOcclusal devices can be either conventionally processed, milled, or printed. However, little is known about the biocompatibility of 3D printing resin materials. PurposeThe purpose of this in vitro study was to compare the viability and morphology of human gingival fibroblast cells (HFG-1) after cultivation on conventionally processed, milled, and printed occlusal device materials with different surface treatments. Material and methodsDisks of a conventionally processed (PalaXpress Clear [pP]), milled (Yamahachi PMMA Clear [sY]), and 2 different printed materials (Dental LT Clear Resin [aD]; Freeprint splint [aF]) were prepared. The surfaces of the specimens were finished by using 2 different treatments (unpolished and polished with P1200-grit silicon carbide paper). HGF-1 cells were cultivated on the specimens for 24 hours, and a viability assay was performed by using polystyrene disks as a control (n=9 disks per group). Cell morphology and the topography of the specimens were examined with scanning electron microscopy (n=3 disks per group). Two-way analysis of variance was applied to determine the effect of material and surface treatment followed by the post hoc Fisher least significant difference test (α=.05). ResultsOverall, material (P<.001) and surface treatment (P<.001) significantly influenced the viability of HGF-1 cells. The viability of cells on all specimens displayed mean values between 0.85 and 1.01 compared with the control except for unpolished aD (0.00 ±0.07) and aF (0.02 ±0.05) that had only a few cells with a round shape. ConclusionsThe behavior of HGF-1 cells on conventionally processed and milled specimens was similar and not dependent on the surface treatment. Unpolished printed specimens had a cytotoxic effect. However, after polishing, cell behavior was similar to that of the conventionally processed and milled specimens.

Influence of dual-cure and self-cure abutment cements for crown implants on human gingival fibroblasts biological properties

AimsTo analyze the biological effects of the cements Relyx Unicem 2, Panavia V5, Multilink Hybrid Abutment and SoloCem on human gingival fibroblast cells (HGFs). Materials and methodsHGFs were exposed to different eluates (n = 40) of the studied resin-based cements. Their cytotoxic effects and influence on cell migration were assessed using MTT and wound-healing assays, respectively. Level of HGF attachment, cell morphology and F-actin cytoskeleton content after exposition to the different eluates were analyzed by scanning electron microscopy (SEM) and confocal microscopy analysis, respectively. The levels of intracellular reactive oxygen species (ROS) produced by the eluates of the different cements were also determined by flow cytometry. Data were analyzed by one-way analysis of variance (ANOVA) followed by Tukey´s test. ResultsEluates of SoloCem significantly reduces the viability of HGFs (69% reduction compared to control at 48 h). Cell migration of HGFs in presence of undiluted SoloCem eluates was significantly lower than in the control (88% open wound area at 24 h). Contrarily, migration speed with Multilynk eluates was similar to that of the control group at all periods of time and all dilutions studied. SEM analysis showed very few cells in SoloCem group, and a moderate cell growth in Multilink, Panavia and Relyx groups were detected. Finally, ROS levels detected in HGFs treated with the more concentrated SoloCem and Relyx dilutions were significantly enhanced compared with that in the control cells or the other groups (44% and 11% ROS positive cells, respectively). ConclusionsThe results obtained in the present work suggest that Multilink hybrid abutment has better biological properties and lower cytotoxicity for cementing implant crowns on abutments.

Comparative evaluation of the cytotoxic effect of different intracanal medicaments

ABSTRACTIntroduction: intracanal medicaments have an important role in disinfecting ofthe pulp space and in endodontic regenerative procedures. These medications comein close contact with the periradicular tissue, thus their degradation products must bebiocompatible with the periapical tissue. Otherwise, these medicaments will resultin intense inflammatory reaction. Aim of the study: The purpose of this in vitrostudy was to evaluate and compare the cytotoxicity of three intracanal medications;Neem oil, double antibiotic paste (DAP) and calcium hydroxide (Ca(OH)2).Materials and Methods: Immortalized human gingival fibroblast cells were retrievedfrom the cell bank and then cultured. Three different intracanal medications (Neemoil, DAP, Ca(OH)2) were added to the cultured cells. Cell viability was evaluatedusing 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) andApoptosis/necrosis assays. Statistical analysis was performed using One-way ANOVAand post-hoc Tukey test. Results: Ca(OH)2 recorded the highest significant cytotoxicityfollowed by Neem oil, then DAP which recorded the lowest significant cytotoxicity.Cytotoxicity of the three intracanal medicaments used were directly proportional totheir concentrations Conclusion: DAP could be better recommended as intracanalmedicament in regenerative endodontic procedures because of its biocompatibilitycompared to the other tested medications.

Cytotoxic Evaluation of Mala aluation of Malaysian K ysian Kelulut Hone elulut Honey on Human y on Human Gingival Fibr al Fibroblast Cell Line using M oblast Cell Line using MTT Assay

Kelulut honey or stingless bee honey is a type of honey produced by stingless bees of the Trigona species where the nest is found in living trees. Objective: The aim of this study was to evaluate the cytotoxic potential of Malaysian Kelulut honey by employing MTT assay on a human gingival fibroblast cell line. Methods: Human gingival fibroblast cell line was cultured in minimal essential medium alpha (α-MEM) with 10% foetal bovine serum and 1% penicillin-streptomycin solution in a 5% CO2 incubator at 37°C in a humidified atmosphere. The cells were seeded at a cell density of 5x103 cells/well in a 96-well culture plate for 24 hours. The cells were treated with seven different concentrations (200, 100, 50, 25, 12.5, 6.25, and 3.125mg/ml) of Malaysian Kelulut honey and incubated in a CO2 incubator. The negative control comprised cells treated with growth media alone. The cell viability was assessed using MTT assay at 24, 48, and 72 hours. The test plate was shaken using a microplate shaker and the absorbance of the solution was measured at 570nm using an ELISA reader with the Magellan software. Statistical analysis of the data was carried out using Kruskal-Wallis test and SPSS 24.0.0 for Windows. A p value &lt;0.05 was considered as statistically significant. Results: There was no cytotoxic effect of Malaysian Kelulut honey on HGF-1 based on the MTT assay at different concentrations and at different time points tested as the cell viability was above 70%. The highest percentage of cell viability at all three different durations of treatment were observed at 3.125mg/ml, whereas the lowest cell viability was observed at 200mg/ml of Kelulut honey concentration. However, statistically significant differences were seen between some of the concentrations at various time points. Conclusion: Since the cell viability of HGF-1 treated with Malaysian Kelulut honey was more than 70% at all concentrations ranging from 3.125mg/ml to 200mg/ml at three different time points (24, 48 and 72 hours), Malaysian Kelulut honey can be considered as non-cytotoxic on human gingival fibroblasts based on MTT assay under the present test conditions

Peptoids with Antibiofilm Activity against the Gram Negative Obligate Anaerobe, Fusobacterium nucleatum.

Peptoids (oligo N-substituted glycines) are peptide analogues, which can be designed to mimic host antimicrobial peptides, with the advantage that they are resistant to proteolytic degradation. Few studies on the antimicrobial efficacy of peptoids have focused on Gram negative anaerobic microbes associated with clinical infections, which are commonly recalcitrant to antibiotic treatment. We therefore studied the cytotoxicity and antibiofilm activity of a family of peptoids against the Gram negative obligate anaerobe Fusobacterium nucleatum, which is associated with infections in the oral cavity. Two peptoids, peptoid 4 (NaeNpheNphe)4 and peptoid 9 (NahNspeNspe)3 were shown to be efficacious against F. nucleatum biofilms at a concentration of 1 μM. At this concentration, peptoids 4 and 9 were not cytotoxic to human erythrocytes or primary human gingival fibroblast cells. Peptoids 4 and 9 therefore have merit as future therapeutics for the treatment of oral infections.

Effect of Leukocyte-Platelet Rich Fibrin (L-PRF) on Tissue Regeneration and Proliferation of Human Gingival Fibroblast Cells Cultured Using a Modified Method.

An in vitro study on rapid culturing method of human gingival fibroblast cells (HGFCs) was established to investigate the potential use of the leukocyte-platelet rich fibrin (L-PRF) in tissue engineering technology, different medical fields, including periodontology and implantology. Eight biopsies were obtained from eight different donors and a modified culturing technique was developed to obtain HGFCs. The modified 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide MTT assay was used to compare the cell viability when the modified culturing method was used in comparison to the standard method. Blood samples were collected from the same patients and L-PRF was isolated using a standard protocol. The releases of platelet-derived growth factor-AA and transforming growth factor-beta1 at various time intervals were observed using enzyme-linked immunosorbent assay (ELISA) kit. The proliferative effect of L-PRF on HGFCs was assessed by the cell counting kit-8 assay. A simple and rapid modified method for in vitro HGFC culture yielded a cellular monolayer within three to nine days after cell culture. L-PRF with three-dimensional polymer fibers released growth factors that peaked during the first three hours and continued to produce up to 10days. The L-PRF presented a dose-dependent effect on HGFCs proliferation where HGFCs proliferation increased with an increase in L-PRF concentration. The modified technique for the culture of HGFCs might be useful for the development of future experimental and clinical studies, besides L-PRF has great therapeutic potential in oral surgery fields.