Human Gingival Fibroblast Cell Lines Research Articles

Examination of cytotoxic and antimicrobial effect of whitening toothpastes: an in vitro study.

Toothpastes are widely used to protect oral and teeth health. This study aims to examine the cytotoxic and antimicrobial effects of whitening toothpastes. In this study, extracts were prepared according to ISO 10993-12:2021 standard (0.2 g/mL) using whitening and conventional toothpastes. The prepared extracts were added to human gingival fibroblast cell lines (HGF-1) in different dilutions (1:1, 1:2, 1:4, 1:8, 1:16, and 1:32) and a cytotoxicity test was performed. Antimicrobial analysis of toothpastes was performed on Streptococcus mutans, Staphylococcus aureus, and Candida albicans using the hole-plate diffusion method. Cell viability and microbial analysis data were examined using two-way analysis of variance (ANOVA) and Tukey post-hoc test (p < 0.05). Toothpastes with sodium lauryl sulfate (SLS) in their composition showed statistically more toxic effects (p < 0.05). The activated carbon toothpastes without SLS showed over 90% cell viability after dilution. Although the dilution rate of toothpastes containing SLS increased, cell viability remained below 70%. All toothpastes used in the study showed antimicrobial effects on S. mutans, S. aureus, and C. albicans. Toothpaste containing hydrogen peroxide and SLS produced more antibacterial effects than activated carbon, blue covarine, microparticles, and conventional toothpaste. SLS-containing toothpastes showed more toxicity on HGF-1 cells. Toothpaste containing hydroxyapatite did not show toxic effects on HGF-1 cells. SLS, sodium lauryl sarcosinate and hydrogen peroxide in toothpastes increase antimicrobial effects.

Insights into the antioxidant, anti-inflammatory and anti-microbial potential of Nigella sativa essential oil against oral pathogens

Oral disorders can exert systemic ramifications beyond their localized effects on dental tissues, implicating a wide array of physiological conditions. The utilization of essential oils (EOs) for protection of oral health represents a longstanding practice. Consequently, in this investigation, essential oil derived from Nigella sativa seeds (NSEO) underwent isolation via the hydro-distillation process, followed by a comprehensive evaluation of its antioxidant, anti-inflammatory, anti-fungal, antibacterial activities, and cytocompatibility. The isolated NSEO manifested as a pale-yellow substance and was found to harbor a diverse spectrum of bioactive constituents, including steroids, triterpenoids, flavonoids, phenols, proteins, alkaloids, tannin, sesquiterpenoid hydrocarbons, monoterpenoid alcohol, and monoterpenoid ketone (thymoquinone). Notably, the total phenolic content (TPC) and total flavonoid content (TFC) of NSEO were quantified at 641.23 μg GAE/gm and 442.25 μg QE/g, respectively. Furthermore, NSEO exhibited concentration-dependent inhibition of protein denaturation, HRBC membrane stabilization, and hemolysis inhibition. Comparative analysis revealed that NSEO and chlorhexidine (CHX) 0.2% displayed substantial inhibition of hemolysis compared to aspirin. While NSEO and CHX 0.2% demonstrated analogous antibacterial activity against Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa, NSEO showcased heightened efficacy against Lactobacillus acidophilus and Candida albicans. Additionally, NSEO exhibited pronounced effects against periodontal pathogens such as Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, and Prevotella intermedia. Importantly, no cytotoxicity was observed on human gingival fibroblast cell lines. These findings underscore the potential of NSEO as a potent antibacterial and antifungal agent in the management of oral microbial pathogens, thereby offering avenues for the development of innovative therapies targeting diverse oral inflammatory conditions. Nevertheless, further investigations are imperative to unlock its full therapeutic repertoire.

Changes in cuproptosis-related gene expression in periodontitis: An integrated bioinformatic analysis

Periodontitis causes inflammatory destruction of tooth-supporting tissues; however, the complex mechanism underlying its etiology remains unclear. Cuproptosis is a type of cell death caused by an imbalance in intracellular copper homeostasis that leads to excess copper. However, changes in the expression and biological function of cuproptosis-related genes (CRGs) in periodontitis are not yet fully understood. This study investigated the comprehensive effects of differentially expressed CRGs (DE-CRGs) on periodontitis via bioinformatic analysis. Nine DE-CRGs were discovered using normal and periodontitis gingival samples, and single-cell RNA sequencing data were analyzed to identify them changes in diverse cell clusters. We then detected the correlation between DE-CRGs and immune infiltration, immune factors, mitochondrial dysfunction, diagnostic efficacy, and predicted drugs. Moreover, changes of DE-CRG in whole periodontitis tissue and a human gingival fibroblast cell line (HGF-1) were confirmed and copper content changes in HGF-1 cells were investigated. Most DE-CRG expression trends were reversed between the periodontal tissues and cell clusters, which may be related to the proportion of cell clusters changes caused periodontitis. Furthermore, most DE-CRG trends in periodontitis cell clusters were inconsistent with the effects of cuproptosis. In HGF-1 cells treated with Porphyromonas gingivalis lipopolysaccharide (Pg-LPS), the intracellular copper content increased by more than threefold, indicating that although some periodontitis cells had excess copper, the amount may not have been sufficient to trigger cuproptosis. Additionally, DE-CRGs were closely associated with multiple biological functions, antibiotic drugs, and natural herbal medicines. Our findings may provide an overview of DE-CRGs in the pathogenesis and treatment of periodontitis.

Non-Invasive Physical Plasma Reduces the Inflammatory Response in Microbially Prestimulated Human Gingival Fibroblasts.

Non-invasive physical plasma (NIPP), an electrically conductive gas, is playing an increasingly important role in medicine due to its antimicrobial and regenerative properties. However, NIPP is not yet well established in dentistry, although it has promising potential, especially for periodontological applications. The aim of the present study was to investigate the effect of NIPP on a commercially available human gingival fibroblast (HGF) cell line and primary HGFs in the presence of periodontitis-associated bacteria. First, primary HGFs from eight patients were characterised by immunofluorescence, and cell numbers were examined by an automatic cell counter over 5 days. Then, HGFs that were preincubated with Fusobacterium nucleatum (F.n.) were treated with NIPP. Afterwards, the IL-6 and IL-8 levels in the cell supernatants were determined by ELISA. In HGFs, F.n. caused a significant increase in IL-6 and IL-8, and this F.n.-induced upregulation of both cytokines was counteracted by NIPP, suggesting a beneficial effect of physical plasma on periodontal cells in a microbial environment. The application of NIPP in periodontal therapy could therefore represent a novel and promising strategy and deserves further investigation.

The Apoptotic Activity of Curcumin Against Oral Cancer Cells Without Affecting Normal Cells in Comparison to Paclitaxel Activity.

Until now, chemotherapy, which has a series of side effects, has been the most widely employed treatment for different types of cancer. However, bioactive products have been utilized as alternative medicines for tumors due to their bioactivities with low or no side effects in normal cells. This research reported for the first time that curcumin (CUR) and paclitaxel (PTX) have significant anti-cancer activity against normal human gingival fibroblast (HGF) and tongue squamous cell carcinoma fibroblast (TSCCF) cell lines. The results showed that CUR (13.85µgmL-1) and PTX (8.17µgmL-1) significantly inhibited TSCCF cell viability, with no significant effect on normal HGF cells. SEM showed morphological changes in cells treated with CUR and PTX, especially with TSCCF cells, compared to HGF normal cells. For TSCCF, the results showed the highest necrosis was achieved with CUR (58.8%) and PTX (39%) as compared to the control (2.99%). For normal HGF cells, the highest early and late apoptosis was achieved with PTX. Further, DCFH-DA analyses showed no significant ROS stimulation in TSCCF and HGF cell lines treated with CUR and PTX. The 1H NMR analysis results show the presence of methoxy and hydroxyl groups and aromatic hydrogens in the CUR structure. In conclusion, the results confirmed that CUR is more specific to the oral cancer cells but not normal cells by inducing apoptosis in a dose- and time-dependent manner, with decreased TSCCF cell viability, and the cytotoxicity of CUR and PTX is not through the ROS pathway.

Evaluation of bioactivity and biodegradability of a biomimetic soft tissue scaffold for clinical use: An in vitro study.

Autogenous soft-tissue graft is the gold-standard approach to augment oral soft tissues. However, tissue engineering is increasingly surveyed to overcome its substantial drawbacks, including the secondary site of operation, patient's pain and discomfort, limited tissue of donor site, and so on. Chitosan and gelatin have been utilized in this field over the years due to their great biological virtues. Zeolite, another remarkable candidate for tissue engineering, possesses outstanding biological and mechanical properties, thanks to its nanostructure. Therefore, this study aimed to investigate the biodegradability and DNA content of seeded human gingival fibroblasts on a New Chitosan-Gelatin-Zeolite Scaffold for the perspective of oral and mucosal soft tissue augmentation. DNA contents of the human gingival fibroblast cell line (HGF.1) seeded on the chitosan-gelatin (CG) and CGZ scaffolds were evaluated by propidium iodide staining on days 1, 5, and 8. Scaffolds' biodegradations were investigated on days 1, 7, 14, 28, 42, and 60. Although both scaffolds provided appropriate substrates for HGF.1 growth, significantly higher DNA contents were recorded for the CGZ scaffold. Among experimental groups, the highest mean value was recorded in the CGZ on day 8. CGZ showed a significantly lower biodegradation percentage at all time points. The incorporation of zeolite into the CG scaffold at a ratio of 1:10 improved the cell proliferation and stability of the composite scaffold. CGZ scaffold may offer a promising alternative to soft-tissue grafts due to its suitable biological features.

Comparison of the Cytotoxic Effect of 3D-Printed Resins, Resin-based CAD/CAM blocks and Composite Resin.

This study investigated the cytotoxic effects of 3D-printed permanent resins, resin-based CAD/CAM blocks and composite resin on human gingival fibroblast (HGF-1) and mouse fibroblast (L929) cell line. 3D-printed permanent resins (Crowntec and Permanent Crown), resin-based CAD/CAM blocks (Vita Enamic and Brilliant Crios) and composite resin (Clearfill Majesty Posterior) were used in the study. Samples were prepared from the planned materials and kept in DMEM according to ISO 10993-12:2021 standard (3 cm2/ml). The cytotoxic effect of the materials on HGF-1 and L929 cells was examined by MTT test at the end of 24 and 72 h. Two-way analysis of variance test (ANOVA) was used to analyze cell viability data. 3D-printed permanent resins, resin-based CAD/CAM blocks and composite resin extracts showed similar cell viability on HGF-1 and L929 cells at the end of 24 h (p>0.05). Resin-based CAD/CAM block (Vita Enamic) produced the highest cell viability on HGF-1 and L929 cells at the end of 72 h (p<0.05). Cell viability values of samples produced in 3D printers with different printing properties did not differ significantly (p>0.05). 3D-printed permanent restoration resins showed similar cell viability on HGF-1 and L929 cells to resin-based CAD/CAM blocks and composite resin. Key words:3D-printed resin, CAD/CAM block, Composite resin, Cytotoxicity, Human Gingival Fibroblast.

Tackling Antibiotic Resistance: Influence of Aliphatic Branches on Broad-Spectrum Antibacterial Polytriazoles against ESKAPE Group Pathogens.

One of the most important threats to public health is the appearance of multidrug-resistant pathogenic bacteria, since they are the cause of a high number of deaths worldwide. Consequently, the preparation of new effective antibacterial agents that do not generate antimicrobial resistance is urgently required. We report on the synthesis of new linear cationic antibacterial polytriazoles that could be a potential source of new antibacterial compounds. These polymers were prepared by thermal- or copper-catalyzed click reactions of azide and alkyne functions. The antibacterial activity of these materials can be modulated by varying the size or nature of their side chains, as this alters the hydrophilic/hydrophobic balance. Antibacterial activity was tested against pathogens of the ESKAPE group. The P3TD polymer, which has butylated side chains, was found to have the highest bactericidal activity. The toxicity of selected polytriazoles was investigated using human red blood cells and a human gingival fibroblast cell line. The propensity of prepared polytriazoles to induce resistance in certain bacteria was studied. Some of them were found to not produce resistance in methicillin-resistant Staphylococcus aureus or Pseudomonas aeruginosa. The interaction of these polytriazoles with the Escherichia coli membrane produces both depolarization and disruption of the membrane.

Antioxidant, Immunomodulatory and Potential Anticancer Capacity of Polysaccharides (Glucans) from Euglena gracilis G.A. Klebs

The present study was carried out to determine the bioactivity of polysaccharides extracted from Euglena gracilis (EgPs). These were characterized by FT-IR and GC-MS. Cytotoxicity analyses (MTT) were performed on healthy human gingival fibroblast cell lines (HGF-1), obtaining an IC50 of 228.66 µg mL-1, and cell lines with anticancer activity for colon cancer (HCT-116), breast cancer (MCF-7), human leukemia (U-937, HL-60) and lung cancer (NCl-H460), showing that EgPs have anticancer activity, mainly in HTC-116 cells (IC50 = 26.1 µg mL-1). The immunological assay determined the immunomodulatory capacity of polysaccharides for the production of proinflammatory cytokines IL-6 and TNF-α in murine macrophages (RAW 264.7) and TNF-α in human monocytes (THP-1). It was observed that the EgPs had a stimulating capacity in the synthesis of these interleukins. The antioxidant capacity of polysaccharides and their biomass were analyzed using the ABTS method (18.30 ± 0.14% and (5.40 ± 0.56%, respectively, and the DPPH method for biomass (17.79 ± 0.57%). We quantitatively profiled HGF-1 proteins by liquid chromatography-tandem mass spectrometry analysis, coupled with 2-plex tandem mass tag labelling, in normal cells. In total, 1346 proteins were identified and quantified with high confidence, of which five were considered to be overexpressed. The data is available through ProteomeXchange, under identifier PXD029076.

Comparative study of effective antibiofilm activity of beneficial microbes-mediated zirconia nanoparticles.

In the present study, beneficial microbes-mediated zirconia nanoparticles were prepared using endophytic bacteria isolated from the seed of Terminalia chebula which were evaluated on inhibition of bacterial adherence and promotion to exhibit antibiofilm properties. The structureand distribution of the zirconia nanoparticles were examined through SEM (Scanning Electron Microscopy), EDS (Energy-Dispersive X-Ray spectroscopy), and XRD (X-ray diffraction analysis), which reveal the distribution of the particles. The morphology of biogenic zirconia nanoparticles was monoclinic and cubic. The formation of zirconia particle was confirmed using UV spectrum and the functional groups were intensified in FTIR (Fourier-transform infrared spectroscopy). The antibiofilm activity of the synthesized nanoparticles was tested in oral pathogens that cause biofilm by membrane integrity and leads to periodontal associated disease. The results showed that the particles had a significant effect on biofilm removal caused by oral pathogens. For determined concentration, the cytotoxicity of the endophytic bacterial facilitated zirconia nanoparticle (Zr NPs) was examined in HGF (Human gingival fibroblast cell line).

Bioactivity of Ionic Liquids Based on Valproate in SH-SY5Y Human Neuroblastoma Cell Line

The search for alternative and effective therapies to fight cancer is one of the main goals of the pharmaceutical industry. Recently, ionic liquids (ILs) have emerged as potential therapeutic agents with antitumor properties. The goal of this study was to synthesize and evaluate the bioactivity of different ILs coupled with the active pharmaceutical ingredient (API) valproate (VPA) as an antitumor agent. The toxicity of the prepared ionic liquids was evaluated by the MTT cell metabolic assay in human neuroblastoma SH-SY5Y and human primary Gingival Fibroblast (GF) cell lines, in which they showed inhibitory effects during the study period. In addition, low cytotoxicity against GF cell lines was observed, suggesting that these compounds are not toxic to human cell lines. [C2OHDMiM][VPA] demonstrated an outstanding antitumor activity against SH-SY5Y and lower activity against the non-neoplastic GF line. The herein assessed compounds played an important role in the modulation of the signaling pathways involved in the cellular behavior. This work also highlights the potential of these ILs-API as possible antitumor agents.

Effects of Litsea cubeba Essential Oil Incorporated into Denture Soft Lining Materials.

The antimicrobial properties, cell cytotoxicity and surface hardness of soft lining materials (GC soft liner, Viscogel and Coe comfort) incorporated with various concentrations of Litsea cubeba essential oil (LCEO) were evaluated. The minimum inhibitory concentrations of LCEO against Candida albicans and Streptococcus mutans were 1.25% v/v and 10% v/v, respectively. However, when LCEO was incorporated into the three soft lining materials (GC soft liner, Viscogel and Coe comfort), 10% v/v and 30% v/v of LCEO could inhibit the growth of C. albicans and S. mutans, respectively. The extracts of soft lining materials with 10% and 30% v/v LCEO, 2% chlorhexidine, 30% v/v nystatin and no additive were used for cytotoxicity tests on a human gingival fibroblast cell line. There was no significant difference in cell viability in all groups with additives compared to the no additive group (p > 0.05). Surface hardness increased significantly between 2 h and 7 day incubation times in all groups, including the controls (p < 0.05). A higher LCEO concentration had a dose-dependent effect on the surface hardness of all soft lining materials (p < 0.05). However, the surface hardness of materials with additive remained in accordance with ISO 10139-1. LCEO could be used as a natural product against oral pathogens, without having a negative impact on soft lining materials.

Phytochemical analysis and cytotoxicity of Clinacanthus nutans on human gingival fibroblast cell line

Introduction and Aim: Clinacanthus nutans (C. nutans) possesses antimicrobial, anticancer, and anti-inflammatory activities. The present study aimed to carry out phytochemical analyses of aqueous and ethanolic extracts of C. nutans leaves and also its cytotoxic potential on human gingival fibroblast cell line HGF-1. Materials and Methods: Phytochemical analyses was conducted using Gas Chromatography Mass Spectrometry (GCMS). Cytotoxicity testing was carried out by treating the HGF-1 with different concentrations (50,100, 200, 400, 800, 1600, 3200, 6400 µg/ml) of both the aqueous and ethanolic extracts of C. nutans leaves using MTT assay. The control comprised only the cell suspension and culture medium. Results: Phytochemical study revealed 53 compounds. Furfural, 2-furanmethanol, butyrolactone, phenol, benzeneacetaldehyde, 4H-Pyran-4-one,2,3-dihydro-3,5-dihydroxy-6-methyl-, vitamin E, campesterol, stigmasterol,.gamma.-Sitosterol and lupeol were the compounds identified in both extracts. Both the extracts did not exhibit any cytotoxicity on HGF-1 based on MTT assay with IC50 values of 1099 µg/ml (ethanol extract) and 2627 µg/ml (aqueous extract). Conclusion: This study showed that ethanol and aqueous extracts of C. nutans leaves contained varied phytochemical compounds and were non-cytotoxic on human gingival fibroblast cell line under the present test conditions.

Maleimide-functionalized carboxymethyl cellulose: A novel mucoadhesive polymer for transmucosal drug delivery

The objective of this research was to develop a novel mucoadhesive polymer for drug delivery applications based on N-(2-aminoethyl) maleimide-functionalized carboxymethyl cellulose in which the weight ratios of the materials were tuned to explore the condition providing the highest maleimide content on the polymer. The polymers were synthesized from N-(2-aminoethyl) maleimide that was conjugated to carboxymethyl cellulose with their mucoadhesive properties examined by tensile testing, rheology, and flow-through analysis and their biocompatibilities evaluated on the human gingival fibroblast cell line (HGF-1). The anti-inflammatory drug benzydamine was loaded into mucoadhesive-polymer-based tablets and used to demonstrate the application of the synthesized polymer. The polymer exhibited superior mucoadhesive capability compared to carboxymethyl cellulose through the interaction between maleimide moiety and mucin. The functionalized polymer also possessed the ability to control the release of benzydamine with Higuchi's release model and was proven to be a potential candidate in mucoadhesive drug delivery.

The efficiency of natural wound healing and bacterial biofilm inhibition of Aloe vera and Sodium Chloride toothpaste preparation

BackgroundPrevention is a preliminary focus of periodontitis treatment. Rather than giving complicated treatment to a periodontitis patient, a variety of toothpastes have been suggested to prevent periodontal disease progression. Herbal toothpastes containing natural plant components for maintaining or increasing healing might be a treatment modality for improving oral hygiene. Aloe vera is a medicinal plant with active ingredients that have antioxidant and anti-inflammatory effects. Additionally, increased sodium in the environment inhibits microorganism growth. A toothpaste containing salt and aloe vera may be an option to provide good oral hygiene.AimTo assess the in vitro cell migration of human gingival fibroblasts and antimicrobial effects of an herbal toothpaste containing A. vera and Sodium chloride.MethodsThe cytotoxicity of 0.02% or 0.2% toothpaste solution on human gingival fibroblast cell line was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The cell migration after treatment with 0.2% (v/v) toothpaste was determined using a Boyden Chamber assay. The effect of the toothpaste on inhibiting Porphylomonas gingivalis planktonic and biofilm growth was compared with Chlohexidine (CHX) using a Disk Diffusion and Biofilm susceptibility test, respectively. The results of the cytotoxicity assay, inhibition zone and percentage of live cells in the biofilm were statistically analyzed with One-way analysis of variance. Cell migration and biofilm inhibition were evaluated using the independent sample t-test and multiple t-test, respectively (p = 0.05).ResultsNeither test concentration of the toothpaste solution was toxic to the target cells. The 0.2% concentration was selected for the cell migration experiment. The herbal toothpaste formulation significantly increased cell migration compared with the control group (culture medium) (p = .02) The antimicrobial effect of this formulation on the P. gingivalis planktonic form was lower compared with 0.12% CHX (positive control group), however, it demonstrated greater P. gingivalis biofilm formation inhibition compared with the 0.12% CHX group.ConclusionsThe alternative use of an herbal toothpaste instead of a non-herbal toothpaste formulation should be considered for promoting oral health care. However, further clinical studies are necessary before it can be considered for patient use.

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.

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.

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

Enhanced Antibiofilm Activity of Endophytic Bacteria Mediated Zirconium Nanoparticles

Unraveling the biofilm formation of oral pathogens by endophytic bacteria-mediated zirconia nanoparticles promotes to exhibits as a dental restorative agent by enhancing antibiofilm activity. To demonstrate the mechanistic studies of biogenic zirconium nanoparticle restricts the growth of oral pathogens. The endophytic bacteria isolated from Terminalia chebula have valuable secondary metabolites used as reducing agents for the synthesis of zirconia nanoparticles. Characterization studies were done for the application of dental restorative material. The nanospheres' shape and size were confirmed by SEM/EDAX followed by XRD and FTIR for their chemical groups that contribute as the antagonist for biofilm. Parallelly evaluation of antioxidant, antibacterial, anti-inflammatory, and antibiofilm activity is opposing the disease-causing pathogens and examined for biocompatibility on Human Primary Gingival fibroblast cell lines. Therefore, the endophytic bacteria mediated zirconia nanoparticles were biologically assessed against oral pathogens, promoting results to exhibit dental restorative biomaterial.