Human Gingival Fibroblast Cells Research Articles

Antimicrobial efficacy of direct air gas soft jet plasma for the in vitro reduction of oral bacterial biofilms

The aim of this study was to evaluate the antimicrobial efficacy of an air gas soft jet CAP for its potential use in removing oral biofilms, given that plasma-based technologies have emerged as promising methods in periodontology. Two types of biofilms were developed, one by Streptococcus mutans UA 159 bacterial strain and the other by a complex mixture of saliva microorganisms isolated from a patient with periodontitis. This latter biofilm was characterized via Next Generation Sequencing to determine the main bacterial phyla. The CAP source was applied at a distance of 6 mm for different time points. A statistically significant reduction of both CFU count and XTT was already detected after 60 s of CAP treatment. CLSM analysis supported CAP effectiveness in killing the microorganisms inside the biofilm and in reducing the thickness of the biofilm matrix. Cytotoxicity tests demonstrated the possible use of CAP without important side effects towards human gingival fibroblasts cell line. The current study showed that CAP treatment was able to significantly reduce preformed biofilms developed by both S. mutans and microorganisms isolated by a saliva sample. Further studies should be conducted on biofilms developed by additional saliva donors to support the potential of this innovative strategy to counteract oral pathogens responsible for periodontal diseases.

Evaluation of Biological Response Elicited by Two Novel Tooth Cream Formulations of Cocos nucifera- Cell Line Studies and MTT Assay on Human Gingival Fibroblast

The current research endeavors to assess the biological reactions induced by two newly formulated tooth creams from Cocos nucifera. Freshly harvested coconut from the Tirupur district of Tamil Nadu underwent processing to create two distinct formulations: one utilizing coconut milk and the other lyophilized coconut extract. Tooth cream samples were prepared and tested against commercial tooth cream following ISO 10993-5 recommendations. Human gingival fibroblast cells were isolated and cultured according to approved protocols, and cytotoxicity evaluations were conducted through MTT assay and live/dead staining. Results indicated high cell viability in both coconut-based formulations, comparable to the commercial tooth cream. Live/dead staining revealed predominantly live cells with minimal cytotoxic effects. Novel coconut-derived tooth creams exhibit comparable biocompatibility to commercial formulations, showing high cell viability and minimal cytotoxicity with human gingival fibroblasts. This suggests coconut-based tooth creams as safe alternatives for oral care, advocating further investigation into their efficacy and tissue safety. Overall, our findings endorse natural compound utilization in dental care formulations.

Antibiofilm activities of denture cleaning tablets against Streptococcus anginosus - Cytotoxic effects on human gingival fibroblast cells

Antibiofilm activities of denture cleaning tablets against Streptococcus anginosus - Cytotoxic effects on human gingival fibroblast cells

Optimization of toothtablets incorporating Albizia myriophylla Benth. stem extract obtained by eco-friendly microwave-assisted extraction

Albizia myriophylla Benth. stems are composed of the bioactive compound lupinifolin, which has exhibited anti-Streptococcus mutans activity. Extracting this plant for incorporation into oral healthcare products is challenging. This study aimed to optimize the eco-friendly microwave-assisted extraction of A. myriophylla stems. It also optimized toothtablets incorporating A. myriophylla stem extract. First, lupinifolin was isolated, and its structure was determined for use as a biomarker in this study. Next, three extraction parameters—the solvent-to-solid ratio, microwave power, and microwave time—were screened using a one-factor-at-a-time approach before being optimized using a Box–Behnken design. The results showed that the optimized conditions that maximized the extraction yield and lupinifolin content were a solvent-to-solid ratio of 23:1, microwave power of 800 W, and microwave time of 70 s. This extract contained 16.27 ± 0.43 % lupinifolin. The extract exhibited a minimum inhibitory concentration of 32 μg/mL and a minimum bactericidal concentration of 256 μg/mL. The half-maximal inhibitory concentration with human gingival fibroblast cells was 102.70 ± 1.02 μg/mL for the extract and 76.75 ± 1.03 μg/mL for lupinifolin. The extract was solidified using microcrystalline cellulose and colloidal silicon dioxide before being incorporated into the toothtablet formulation. Three factors in toothtablet production—compression force, microcrystalline cellulose content, and hydrated silica content—were also screened using a one-factor-at-a-time approach before being optimized using a Box–Behnken design. The results showed that the optimized conditions that approached the desired properties were a compression force of 2500 psi, microcrystalline cellulose content of 12 %, and hydrated silica content of 10 %. The developed toothtablets contained 35.64 ± 0.45 mg of lupinifolin per 650 mg toothtablet (or approximately 5.5 % lupinifolin per toothtablet) and retained their anti-S. mutans activity. They appeared safe for human gingival fibroblasts, even when treated for 2 min, with a maximum concentration of 2000 μg/mL. In conclusion, the developed toothtablets incorporating A. myriophylla stem extract can potentially serve as a high-quality, antibacterial, and safe alternative to traditional toothpaste.

Investigating the Viability of Human Gingival Fibroblast Cells in a Polyhydroxybutyrate/Chitosan/Bioglass Nanoparticles Scaffold

Background: The primary method for addressing dental problems traditionally involves the removal and replacement of damaged tissue with synthetic substitutes. However, these materials often lack the physical and chemical properties of natural teeth and are prone to mechanical failures. Recent advancements in tissue engineering have highlighted an alternative approach that allows for the regeneration of teeth using scaffolds. Objectives: This study explored the viability of human gingival fibroblast cells in a scaffold composed of polyhydroxybutyrate, chitosan, and bioglass nanoparticles (PHB/chitosan/nBG). Methods: This study is experimental and was conducted in a laboratory setting. Scaffolds of PHB, PHB/chitosan, and PHB/chitosan/nBG were previously prepared from PHB polymer, chitosan, and bioglass using the electrospinning method. Human gingival fibroblast cells (C165) were acquired from the Pasteur Institute and cultured. Twenty thousand cells were seeded onto each scaffold and into the wells of a 24-well plate for periods of 3, 5, and 7 days. Subsequently, cell viability and proliferation were assessed using the MTT assay method, with each experiment repeated three times. Results: Cell viability and proliferation on the PHB/chitosan/nBG scaffold demonstrated a significant increase compared to the other scaffolds (P ≤ 0.01). Conclusions: The PHB/chitosan/nBG scaffold was found to be non-toxic to human gingival fibroblast cells and exhibited good biocompatibility.

Viability and Proliferation Assessment of Gingival Fibroblasts Cultured on Silver Nanoparticle-Doped Ti-6Al-4V Surfaces.

To investigate the biocompatibility of silver nanoparticle (AgNP)-doped Ti-6Al-4V surfaces by evaluating the viability and proliferation rate of human gingival fibroblasts (HGFs)-as the dominant cells of peri-implant soft tissues-seeded on the modified surfaces. AgNPs (sizes 8 nm and 30 nm) were incorporated onto Ti-6Al-4V specimen surfaces via electrochemical deposition, using colloid silver dispersions with increasing AgNP concentrations of 100 ppm, 200 ppm, and 300 ppm. One control and six experimental groups were included in the study: (1) control (Ti-6Al-4V), (2) 8 nm/100 ppm, (3) 8 nm/200 ppm, (4) 8 nm/300 ppm, (5) 30 nm/100 ppm, (6) 30 nm/200 ppm, and (7) 30 nm/300 ppm. HGF cell primary cultures were isolated from periodontally healthy donor patients and cultured in direct contact with the group specimens for 24 and 72 hours. The cytotoxicity of AgNP-doped Ti-6Al-4V specimens toward HGF was assessed by the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) and BrdU (5-bromo-2'-deoxyuridine) assay tests. Calcein AM and ethidium homodimer (EthD-1) fluorescent stains were used to determine the live and dead cells. The morphology and attachment properties of the HGFs were determined via scanning electron microscopy (SEM). Energy dispersive x-ray (EDX) analysis confirmed the presence of AgNPs on the specimens. The MTT test revealed that AgNPs of both sizes and all concentrations presented a decreased cellular metabolic activity compared to the control discs. All concentrations of both sizes of AgNPs affected the cell proliferation rate compared to the control group, as revealed by the BrdU assay. Overall, cytotoxicity of the modified Ti-6Al-4V surfaces depended on cell exposure time. Observation via confocal microscopy confirmed the results of the MTT and BrdU assay tests. Specifically, most cells remained alive throughout the 72-hour culture period. SEM images revealed that adjacent cells form bonds with each other, creating confluent layers of conjugated cells. The findings of the present study indicate that Ti-6Al-4V surfaces modified with 8 nm and 30 nm AgNPs at concentrations of 100 ppm, 200 ppm, and 300 ppm do not produce any serious cytotoxicity toward HGFs. The initial arrest of the HGF proliferation rate recovered at 72 hours. These results on the antibacterial activity against common periodontal pathogens, in combination with the results found in a previous study by the same research group, suggest that AgNP-doped Ti-6Al-4V surfaces are potential candidates for use in implant abutments for preventing peri-implant diseases.

In Vitro Cytotoxicity of Linseed Oil as Bio-Based Plasticiser in Polymethylmethacrylate (PMMA) Denture Base Material

Objectives: Polymethyl methacrylate (PMMA) is a polymer that is vastly utilised as denture base material. Previously synthetic plasticiser like Dibutyl phthalate was previously used in the PMMA. However, there are reports of ester leaching causing intraoral toxicity. Thus, bringing the awareness about organic plasticiser usage, like linseed oil. This study aims to evaluate the cytotoxicity of linseed oil to ensure its safety to be used as the natural-based plasticiser in PMMA as denture base material. Materials and Methods: The cytotoxicity of linseed oil was evaluated by culturing Human Gingival Fibroblast cells with 1, 3, 5, 7, and 10% linseed oil for 24, 48, and 72 hours. The WST-1 assay was then used to evaluate cell viability, with cell quantification performed using a spectrophotometer at a wavelength range of 420-480nm. An unpaired t-test was used to analyse all parameters' quantitative cell viability data. Results: The WST-1 Assay demonstrated that linseed oil exhibited greater than 95% cell viability at all doses and incubation durations, indicating biocompatibility. At 72 hours of incubation, there are statistically significant differences between the cells incubated in the Linseed Oil as compared to Dimethyl Sulfoxide (DMSO), which acts as a positive control. Conclusions: The study proves that linseed oil is non-cytotoxic with the Human Gingival Fibroblast cells and safe to use when incorporated later in the PMMA as denture base material.

Anticancer effects of rutin from Fagopyrum tataricum (tartary buckwheat) against osteosarcoma cell line.

The present study is analysisof the seeds of buckwheat (Fagopyrum sp.),member of the Polygonaceae family for isolation of rutin and its anticancer property againstOsteosarcoma celllines (SAOS2). The selected plant is traditionally used for diabetes and cancer. It has several biological properties such as antibacterial, antioxidant and anti-aging. Thirty-five buckwheat cultivars were obtained from Nepal Agriculture Genetic Resources Centre (NAGRC) Khumaltar, Kathmandu, Nepal, andKumrek Sikkim. These plant varieties are scientifically evaluated their biological properties. Rutin wasfractionated from buckwheat seeds using methanol fraction and analysed for quality by HPLC method. The rutin fraction of the cultivar NGRC03731 a tartary buck wheat and standard rutin was used against Osteosarcoma cell lines (SAOS2) and human gingival fibroblast cells (hGFs) for anticancer activity. The cell viability using rutin fraction and standard rutin treated with SAOS2 cells were assessed by MTT assay. For further research, the best doses (IC-50: 20g/ml) were applied. By using AO/EtBr dual staining, the effects of Rutin fraction on SAOS2 cell death were analysed. The scratch wound healing assay was used to analyse cell migration. Real-time PCR was used to analyse the pro-/anti-apoptotic gene expression. The seeds with the highest rutin content, NGRC03731 seeds, had 433mg/100g of rutin.The rutin fraction treatment and standard rutin significantly reduced cell viability in the MTT assay, and osteosarcoma cells were observed on sensitive to the IC-50 dose at a concentration of 20g/ml after 24h.The SAOS2 cells exposed to rutin fraction at 20g/ml and standard rutin at 10g/ml exhibited significant morphological alterations, cell shrinkage and decreased cell density, which indicate apoptotic cells.Rutin-fraction treated cells stained with acridine orange/ethidium bromide (AO/EtBr) dual staining cells turned yellow, orange, and red which indicatesto measure apoptosis.The anti-migration potential of rutin fraction, results prevented the migration of SAOS2 cancer cells.Rutin-fraction significantly increased the expression of pro-apoptotic proteinsBad, using real-time PCR analysis (mRNA for Bcl-2 family proteins) resulted Bcl-2's expression is negatively regulated. Osteosarcoma (SAOS2) cell lines' proliferation, migration, and ability to proliferate were reduced markedly by rutin fraction and it also causes apoptosis of Osteosarcoma cell lines (SAOS2).

Antibiofilm Activity and Biocompatibility of Temporin-SHa: A Promising Antimicrobial Peptide for Control of Fluconazole-Resistant Candida albicans.

The aim of the study was to investigate the effect of antimicrobial peptides (AMPs) Hylin-a1, KR-12-a5, and Temporin-SHa in Candida albicans as well as the biocompatibility of keratinocytes spontaneously immortalized (NOK-si) and human gingival fibroblasts (FGH) cells. Initially, the susceptible (CaS-ATCC 90028) and fluconazole-resistant (CaR-ATCC 96901) C. albicans strains were grown to evaluate the effect of each AMP in planktonic culture, biofilm, and biocompatibility on oral cells. Among the AMPs evaluated, temporin-SHa showed the most promising results. After 24 h of Temporin-SHa exposure, the survival curve results showed that CaS and CaR suspensions reduced 72% and 70% of cell viability compared to the control group. The minimum inhibitory/fungicide concentrations (MIC and MFC) showed that Temporin-SHa was able to reduce ≥50% at ≥256 µg/mL for both strains. The inhibition of biofilm formation, efficacy against biofilm formation, and total biomass assays were performed until 48 h of biofilm maturation, and Temporin-SHa was able to reduce ≥50% of CaS and CaR growth. Furthermore, Temporin-SHa (512 µg/mL) was classified as non-cytotoxic and slightly cytotoxic for NOK-si and FGH, respectively. Temporin-SHa demonstrated an anti-biofilm effect against CaS and CaR and was biocompatible with NOK-si and FGH oral cells in monolayer.

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.

Comparison of thewound healing effects of vascular endothelial growth factor a (VEGF-A) and transforming growth factor beta 3 (TGF-β3) on gingival cells

Each tissue has its own anatomy and physiology. The deterioration of these structures by various effects (crushing, puncture, pathogens, etc.) is called as wound. The tissue exposed to these effects wants to gain the same function and anatomy. This phenomenon is called wound healing. Wound healing is a process that involves many reactions and cytokines. Since it is known that TGF-β3 and VEGF-A proteins which are among these cytokines, play an active role in wound healing stages. In the present study, it was decided to examine the wound healing effects of the non human gingival fibroblast cells comperatively. Wound healing process under the influence of these cytokines was performed by in vitro scratch analysis which is known as easy, reliable and cheap technique. In this context, firstly cell viability analysis was performed to determine the most effective dose for inducing cell proliferation. After this experimental step, gingival cells were incubated for 72 hours with suitable TGF-β3 and VEGF-A doses. As a result of the study, it was concluded that TGF-β3 and VEGF-A proteins are not cytotoxic and have a wound-healing effect on gingival fibroblast cells.

Evaluating the Translucency, Surface Roughness, and Cytotoxicity of a PMMA Acrylic Denture Base Reinforced with Bioactive Glasses.

The colonisation of the surface of removable acrylic dentures by various types of microorganisms can lead to the development of various diseases. Therefore, the creation of a bioactive material is highly desirable. This study aimed to develop a denture base material designed to release bioactive ions into the oral environment during use. Four types of bioactive glasses (BAG)-S53P4, Biomin F, 45S5, and Biomin C-were incorporated into the PMMA acrylic resin, with each type constituting 20 wt.% (10 wt.% non-silanised and 10% silanised) of the mixture, while PMMA acrylic resin served as the control group. The specimens were subsequently immersed in distilled water, and pH measurements of the aqueous solutions were taken every seven days for a total of 38 days. Additionally, surface roughness and translucency measurements were recorded both after preparation and following seven days of immersion in distilled water. The cytotoxicity of these materials on human fibroblast cells was evaluated after 24 and 48 h using Direct Contact and MTT assays. Ultimately, the elemental composition of the specimens was determined through energy-dispersive X-ray (EDX) spectroscopy. In general, the pH levels of water solutions containing BAG-containing acrylics gradually increased over the storage period, reaching peak values after 10 days. Notably, S53P4 glass exhibited the most significant increase, with pH levels rising from 5.5 to 7.54. Surface roughness exhibited minimal changes upon immersion in distilled water, while a slight decrease in material translucency was observed, except for Biomin C. However, significant differences in surface roughness and translucency were observed among some of the BAG-embedded specimens under both dry and wet conditions. The composition of elements declared by the glass manufacturer was confirmed by EDX analysis. Importantly, cytotoxicity analysis revealed that specimens containing BAGs, when released into the environment, did not adversely affect the growth of human gingival fibroblast cells after 48 h of exposure. This suggests that PMMA acrylics fabricated with BAGs have the potential to release ions into the environment and can be considered biocompatible materials. Further clinical trials are warranted to explore the practical applications of these materials as denture base materials.

New Physico-Chemical Analysis of Magnesium-Doped Hydroxyapatite in Dextran Matrix Nanocomposites.

The new magnesium-doped hydroxyapatite in dextran matrix (10MgHApD) nanocomposites were synthesized using coprecipitation technique. A spherical morphology was observed by scanning electron microscopy (SEM). The X-ray diffraction (XRD) characterization results show hydroxyapatite hexagonal phase formation. The element map scanning during the EDS analysis revealed homogenous distribution of constituent elements of calcium, phosphor, oxygen and magnesium. The presence of dextran in the sample was revealed by Fourier transform infrared (FTIR) spectroscopy. The antimicrobial activity of the 10MgHAPD nanocomposites was assessed by in vitro assays using Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, Streptococcus mutans ATCC 25175, Porphyromonas gingivalis ATCC 33277 and Candida albicans ATCC 10231 microbial strains. The results of the antimicrobial assays highlighted that the 10MgHApD nanocomposites presented excellent antimicrobial activity against all the tested microorganisms and for all the tested time intervals. Furthermore, the biocompatibility assays determined that the 10MgHApD nanocomposites did not exhibit any toxicity towards Human gingival fibroblast (HGF-1) cells.

Dipotassium glycyrrhizate and hinokitiol enhance macrophage efferocytosis by regulating recognition, uptake, and metabolism of apoptotic cells invitro.

Efferocytosis is a process whereby macrophages remove apoptotic cells, such as neutrophils, that have accumulated in tissues, which is required for resolution of inflammation. Efferocytosis is impaired in individuals with increasing age and in those with various systemic diseases. Recently, efferocytosis has been reported to be related to the pathogenesis and progression of periodontitis, and enhancement of efferocytosis, especially in the subjects with impaired efferocytosis, was suggested to lead to periodontitis prevention and care. Various anti-inflammatory ingredients are used in oral care products, but their effect on efferocytosis is unclear. Here, we aimed to identify ingredients contained in oral care products that are effective for efferocytosis regulation. The ability of dead cells to induce inflammation in human gingival fibroblast (HGF) cells were evaluated by measuring IL-6 secretion. Six ingredients in oral care products used as anti-inflammatory agents were evaluated for their effect on efferocytosis using flow cytometry. The expression of various efferocytosis-related molecules, such as MERTK and LRP1 involved in recognition, and LXRα and ABCA1 that function in metabolism, were measured in RAW264.7 cells with or without ingredient treatment. Rac1 activity, which is related to the uptake of dead cells, was measured using the G-LISA kit. Dead cells elicited IL-6 secretion in HGF cells. Among the six ingredients, GK2 and hinokitiol enhanced efferocytosis activity. GK2 and hinokitiol significantly increased the expression of MERTK and LRP1, and also enhanced LXRα and ABCA1 expression after efferocytosis. Furthermore, they increased Rac1 activity in the presence of dead cells. Among the six ingredients tested, GK2 and hinokitiol promoted efferocytosis by regulating apoptotic cell recognition, uptake, and metabolism-related molecules. Efferocytosis upregulation may be one of the mechanisms of GK2 and hinokitiol in the treatment of inflammatory diseases, such as periodontitis.

An evaluation of photobiomodulation effects on human gingival fibroblast cells under hyperglycemic condition: an in vitro study.

An in vitro study was designed to evaluate the effects of photobiomodulation (PBM) with 915-nm diode laser on human gingival fibroblast (HGF) cells under hyperglycemic condition. The HGF cells were cultured in Dulbecco's modified eagle medium (DMEM) medium containing 30mM glucose concentration for 48h to mimic the hyperglycemic condition. Subsequently, the cells received three sessions of PBM (915nm, continuous emission mode, 200 mW, energy density values of 3.2, 6, and 9.2J/cm2). Twenty-four hours post-irradiation, cell proliferation, expression of interleukin 6 (IL-6), and vascular endothelial growth factor (VEGF) were assessed with MTT and real-time polymerase chain reaction (PCR) tests, respectively. Also, reactive oxygen species (ROS) production was measured using CM-H2DCFDA fluorimetry. No changes were detected in the cell proliferation rate between the high glucose control group and laser-treated cells, while VEGF and IL-6 gene expression levels increased significantly after PBM in the high glucose-treated cells group. ROS level was significantly decreased in the irradiated cells in high-glucose medium compared with the high glucose control group. Our study revealed the inductive role of 915-nm-mediated PBM on VEGF and the inflammatory response while concurrently reducing reactive oxygen species production in HGF cells in hyperglycemic conditions.

Targeting Biomechanical Endurance of Dental-Implant Abutments Using a Diamond-Like Carbon Coating.

Abutment components (i.e., fixtures associated with oral implants) are essentially made of titanium (Ti), which is continuously exposed to the hash oral environment, resulting in scratching. Thus, such components need to be protected, and surface treatments are viable methods for overcoming long-term damage. Diamond-like carbon (DLC), an excellent protective material, is an alternative surface-treatment material for Ti abutments. Here, we demonstrate that a silicon interlayer for DLC film growth and the pulsed-direct current plasma-enhanced chemical vapor deposition (DC-PECVD) method enables the deposition of an enhanced protective DLC film. As a result, the DLC film demonstrated a smooth topography with a compact surface. Furthermore, the DLC film enhanced the mechanical (load-displacement, hardness, and elastic modulus) and tribological properties of Ti as well as increased its corrosion resistance (16-fold), which surpassed that of a bare Ti substrate. The biofilm formed (Streptococcus sanguinis) after 24 h exhibited an equal bacterial load (∼7 Log colony-forming units) for both the groups (Ti and DLC). In addition, the DLC film exhibited good cytocompatibility, owing to its noncytotoxicity toward human gingival fibroblast cells. Therefore, DLC deposition via DC-PECVD can be considered to be a promising protective and cytocompatible alternative for developing implant abutments with enhanced mechanical, tribological, and electrochemical properties.

Accelerated Oral Healing by Angelica gigas Nakai from Hot Melt Extrusion Technology: An In Vitro Study.

Background and Objectives: In spite of the oral environment being healing-prone, its dynamic changes may affect wound healing. The purpose of this study was to assess the oral wound healing effect of Angelica gigas Nakai (AG) prepared by hot-melt extrusion. Materials and Methods: Human gingival fibroblast (HGF) cells were treated with AG or AG via hot-melt extrusion (AGH) for 24 h to determine the optimal concentration. For evaluating the anti-inflammatory effect of AG and AGH, a nitric oxide assay was performed under lipopolysaccharide (LPS) stimulation. The wound-healing effects of AG and AGH were evaluated using cell proliferation/migration assays and wound-healing marker expression through qRT-PCR. Results: Both AG and AGH showed no cytotoxicity on HGH cells. Regarding nitric oxide production, AGH significantly decreased LPS-induced nitric oxide production (p < 0.05). AGH showed a significantly positive result in the cell proliferation/cell migration assay compared with that in AG and the control. Regarding wound healing marker expression, AGH showed significantly greater VEGF and COL1α1 expression levels than those in the others (p < 0.05), whereas α-SMA expression was significantly different among the groups. Conclusions: Within the limits of this study, AGH accelerated oral wound healing in vitro.

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.

Passaging of gingival fibroblasts from periodontally healthy and diseased sites upregulates osteogenesis-related genes.

To investigate biological processes of the periodontium, in vitro primary cell models have been established. To study the biology of the gingiva, primary gingival fibroblast cell models are widely used. For such experiments, cells need to be expanded and passaged. A key assumption is that primary cells maintain most of their original characteristics they have in situ. The aim of this research is to explore the impact of early passaging on selected gene expression of human gingival fibroblast cells. For this purpose, gene expression from the outgrowth of the resected tissues until the fourth passage was followed for nine tissue samples, from both healthy and diseased sites. Micrographs were taken from the cultures, RNA was extracted from the samples of each passage and quantitative PCR was performed for selected genes representing various biological processes. Epithelial cells were present during the first outgrowth, but were no longer present in the second passage. Our results indicate that the morphology of the gingival fibroblast cells does not change with passaging and that passages 2-4 contain only gingival fibroblasts. Gene expression of M-CSF, TNF-α, TLR4, POSTN and FAPα was unchanged by passaging, the expression of IL-6, IL-1β and TLR2 decreased due to passaging and the expression of in particular the selected osteogenesis genes (ALP, RUNX2, Osteonectin, COL1A), OPG and MKI67 increased with passaging. Worldwide, use of the same passage in laboratory experiments using primary cell cultures is the standard. Our results support this, since for certain genes, in particular osteogenesis genes, expression may alter solely due to passaging.

Comparative effect of anthocyanin on proliferation and migration of human gingival fibroblasts in the absence or presence of nicotine.

Oral fibroblast malfunction can result in periodontal diseases. Nicotine can prolong the healing process as an irritant of oral tissues. Anthocyanins have been demonstrated to have potential benefits in preventing or treating smoking-related periodontal diseases. Cyanidin chloride's (CC's) potential in oral wound healing and the viability, proliferation, and migration of human gingival fibroblasts (HGFs) were examined in the presence and absence of nicotine by an in vitro study. The effects of different nicotine concentrations (1, 2, 3, 4, and 5 mM) on the viability and proliferation of HGF cells were evaluated in the presence and absence of different CC concentrations (5, 10, 25, and 50 μM) using the quantitative MTT assay. The scratch test was performed to evaluate the migration of CC-treated cells in the presence of 2.5-mM nicotine. No cytotoxicity was observed at 1‒100 μM CC concentrations after 24, 48, and 72 hours of exposure to HGF cells. However, a concentration of 200 μM significantly reduced cell viability by about 20% at all the three-time intervals (P<0.05). Also, 3‒5-mM concentrations of nicotine significantly reduced cell viability in a dose- and time-dependent manner. Moreover, the understudied CC concentrations decreased nicotine's adverse effects on cell migration to some extent. Although the understudied CC concentrations could not significantly reduce the adverse effects of understudied nicotine concentrations on the viability and proliferation of HGF cells, they were able to reduce the detrimental effects of nicotine on cell migration significantly.