HGF-1 Cells Research Articles

Cytocompatibility and Wound Healing Activity of Chitosan Thiocolchicoside Lauric Acid Nanogel in Human Gingival Fibroblast Cells.

To investigate the cytocompatibility effect and wound healing activity of chitosan thiocolchicoside lauric acid (CTL) nanogel using human gingival fibroblast (hGF) cells. hGF cells were established from gingival tissue as per the standard cell isolation protocol. The cytocompatibility effect was assessed using an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide) assay. A scratch wound healing assay was performed to assess the wound-healing potential of CTL nanogel. For the nuclear morphological changes analysis, acridine orange staining was used in gingival fibroblast cells. The stained nuclei were viewed under a fluorescent microscope. ANOVA with posthoc analysis was performed using GraphPad Prism 5 software (Dotmatics, Boston, Massachusetts). The significance level (p-value) was expressed as <0.05. Results: CTL nanogel did not show any significant cytotoxicity at concentrations 10-80 µl/ml (p<0.05). CTL nanogel at a concentration of 40µl/ml has a cytocompatibility effect on hGF cells and increases cell viability. In vitro scratch wound healing assay resulted in faster wound healing and cell migration with CTL nanogel when compared to the control group. CTL nanogel has a significant effect on cell proliferation at various concentrations, which suggests its use as a safe and effective drug delivery system.

Synthesis, Structural Modification, and Bioactivity Evaluation of Substituted Acridones as Potent Microtubule Affinity-Regulating Kinase 4 Inhibitors.

Acridones present numerous pharmacological activities, including inhibition of microtubule affinity-regulating kinase 4 (MARK4) kinase activity. To investigate structure-activity relationships and develop potent MARK4 inhibitors, derivatives of 2-methylacridone were synthesized and tested for their activity against MARK4 kinase. Selective substitutions at the nitrogen atom were accomplished by treating 2-methylacridone with alkyl halides in the presence of K2CO3. In addition, amidation of acridone acetic acid 11 with piperazine or tryptophan methyl ester followed by derivatization with various amines gave a series of new acridone derivatives. Among the tested compounds, six were identified as possessing high inhibitory activity against MARK4. The molecular modeling studies showed that the derivatives bearing piperazine or tryptophan bind well to the ATP-binding site of MARK4. The antiproliferative activity of six active compounds was evaluated against HeLa and U87MG cancer cells. Tryptophan derivatives 23a, 23b, and 23c showed significant cytotoxicity against both cell lines with EC50 values ranging from 2.13 to 4.22 μM, while derivatives bearing piperazine were found to be not cytotoxic. Additionally, compound 23a decreased the proliferation of human MDA-MB-435 and U251 cancer cells in the low micromolar range; however, it also affects the non-cancerous HGF cells. Due to their high binding affinity against MARK4, the synthesized compounds could be potential agents to target MARK4 against cancer and tauopathies.

Determination of cytotoxic, anti-acetylcholinesterase and antioxidant activity of some medicinal Artemisia spp.

The study aims to investigated the antioxidant activities, phenolic compounds, acetylcholinesterase enzyme inhbition and cytotoxic effects of two different of Artemisia spp. (Artemisia dracunculus L. and Artemisia dracunculoides Pursh) cultivated in Turkey organically, for the first time. Total phenolic, total flavonoid contents of the plants determined spectrophotometrically while antioxidant activity DPPH, CUPRAC, and FRAP was determined using colorimetric method. And also acetylcholinesterase enzyme inhibition activity and anti-cancer activity in-vitro aganist human melanoma (VMM917, CRL-3232), lung carcinoma (A549, CCL-185) and, normal human fibroblast (hGF, PCS-201-018) cells were studied. Total phenolic (225-324 mgGAE/g sample) and total flavonoid contents (0.066-0.085 mgQAE/g sample), antioxidant activity (DPPH (1.371-1.655 mg/mL), CUPRAC (0.246-0.344 µM CTEAC) and FRAP (462.133-726.661 µM CTEAC)). A. dracunculus and A. dracunculoides extracts inhibited 40.09±0.65%, and 39.48±3.68% of acetylcholinesterase activity. It was determined that demonstrated the selective effect of A. dracunculus and A. dracunculoides on the cytotoxicity of A549 and hGF cells.

Evaluation of the HGF-1 Cells on Titanium Abutments Fixed by Various Procedures

Bu çalışmanın amacı, titanyum abutment üzerinde, insan gingival fibroblast hücre hattının (HGF-1), tutunma ve proliferasyon açısından fiksasyon solüsyon ve sürelerinin etkilerinin SEM ve İmmunofloresan görüntüleme ile değerlendirilmesi ve kıyaslanmasıdır. Hazır temin edilen insan gingival hücre hattı (HGF-1) 10x10x1cm3 boyutunda 32 adet titanyum alaşım (Ti6Al4V) plaka üzerine ekildi. 8 grup belirlendi (n=4). 48 saat sonucunda hücreler değerlendirildi. Örnekler Gluteraldehit ile 30, 45, 60 dakika (Grup GA30, GA45, GA60), Formaldehit ile 6, 12, 24 saat (Grup FA6, FA12, FA24) ve Paraformeldehit ile 2 saat ve 20 dakika (Grup PFA2, PFA20) süre ile fikse edildi. Fiksasyon sonrası her gruptan 2 örnek Taramalı elektron mikroskobunda (SEM) ve 2 örnek İmmunofloresan mikroskobunda görüntülenmek için hazırlandı. Tüm gruplardaki hücrelerin temas alanları ölçüldü, saatler arasındaki farklılık istatistiksel olarak değerlendirildi. SEM değerlendirmesinde, en uygun HGF-1 hücre morfolojisi görüntülerinin formaldehit gruplarının iğsi şekilli homojen yayılım gösterdiği tespit edildi. FA12 grubunda hücre temas alanı %95,83 bulunmuş olup, tüm deney grupları içerisinde en iyi hücre yayılımını göstermiştir. Gluteraldehit ve paraformaldehit gruplarında birbirleriyle benzer şekilde hücrelerin uzamış, belirli alanlarda öbekleşmiş ve üst üste katmanlanmış belirgin çekirdekli hücre görüntüleri tespit edildi. İmmunofloresan görüntülerinde her üç (gluteraldehit, formaldehit, paraformaldehit) gruptaki hücrelerde de aktin filamentlerinin yoğunlukları benzer seviyelerde görülmesinin yanı sıra paraformaldehit gruplarında titanyum yüzeydeki hücre gövdelerinin diğer fiksatif gruplarına göre daha belirgin, iyi yayılmış ve daha büyük yüzey alanlarına sahip olduğu gözlendi. Fiksasyon hücre çalışmalarında görüntülemenin en kritik basamaklarından biridir. Araştırmacıların başarılı görüntü sonucu elde edebilmek için en uygun fiksasyon yöntemini göz önünde bulundurmaları gerekmektedir.

Optimized Gingiva Cell Behavior on Dental Zirconia as a Result of Atmospheric Argon Plasma Activation.

Several physico-chemical modifications have been developed to improve cell contact with prosthetic oral implant surfaces. The activation with non-thermal plasmas was one option. Previous studies found that gingiva fibroblasts on laser-microstructured ceramics were hindered in their migration into cavities. However, after argon (Ar) plasma activation, the cells concentrated in and around the niches. The change in surface properties of zirconia and, subsequently, the effect on cell behavior is unclear. In this study, polished zirconia discs were activated by atmospheric pressure Ar plasma using the kINPen®09 jet for 1 min. Surfaces were characterized by scanning electron microscopy, X-ray photoelectron spectroscopy (XPS), and water contact angle. In vitro studies with human gingival fibroblasts (HGF-1) focused on spreading, actin cytoskeleton organization, and calcium ion signaling within 24 h. After Ar plasma activation, surfaces were more hydrophilic. XPS revealed decreased carbon and increased oxygen, zirconia, and yttrium content after Ar plasma. The Ar plasma activation boosted the spreading (2 h), and HGF-1 cells formed strong actin filaments with pronounced lamellipodia. Interestingly, the cells' calcium ion signaling was also promoted. Therefore, argon plasma activation of zirconia seems to be a valuable tool to bioactivate the surface for optimal surface occupation by cells and active cell signaling.

Magnolia kobus Extract Suppresses Porphyromonas gingivalis LPS-Induced Proinflammatory Cytokine and MMP Expression in HGF-1 Cells and Regulates Osteoclastogenesis in RANKL-Stimulated RAW264.7 Cells.

Clinical prevention is of utmost importance for the management of periodontal diseases. Periodontal disease starts with an inflammatory response in the gingival tissue, and results in alveolar bone destruction and subsequent tooth loss. This study aimed to confirm the anti-periodontitis effects of MKE. To confirm this, we studied its mechanism of action using qPCR and WB in LPS-treated HGF-1 cells and RANKL-induced osteoclasts. We found that MKE suppressed proinflammatory cytokine protein expression by inhibiting the TLR4/NF-κB pathway in LPS-PG-induced HGF-1 cells and blocking ECM degradation by regulating the expression of TIMPs and MMPs. We also confirmed that TRAP activity and multinucleated cell formation were reduced in RANKL-stimulated osteoclasts after exposure to MKE. These results were confirmed by inhibiting TRAF6/MAPK expression, which led to the suppression of NFATc1, CTSK, TRAP, and MMP expression at the gene and protein levels. Our results confirmed that MKE is a promising candidate for the management of periodontal disease based on its anti-inflammatory effects and inhibition of ECM degradation and osteoclastogenesis.

Aqueous Extract of Sparganii Rhizoma and Curcumae Rhizoma Induces Apoptosis and Inhibits Migration in Human Oral Squamous Cell Carcinoma

Sparganii Rhizoma and Curcumae Rhizoma (SRCR) are natural herbs used in traditional Chinese medicine to treat tumors and activate blood circulation. Previous studies have shown that SRCR possesses notable antitumor activity; however, the mechanism underlying anticancer activity in human oral squamous cell carcinoma (OSCC) has yet to be fully elucidated. The risk factors of OSCC include smoking, alcohol consumption, poor oral hygiene, and human papillomavirus infection. OSCC is highly metastatic and responds poorly to chemotherapy; thus, alternative treatment options are imperative. In this study, we found that SRCR induced death in OSCC cells but not in normal cells (HGF-1 cells). SRCR has also shown to induce the production of reactive oxygen species in OSCC cells, sequentially promoting calcium release and stress in the endoplasmic reticulum, which resulted in mitochondrial dysfunction and subsequent apoptosis. Furthermore, SRCR has shown to inhibit the migration of OSCC cells by reducing matrix metalloproteinase-12 and -13. Our findings demonstrate that SRCR exerts anticancer activities in OSCC by inducing cell apoptosis and suppressing cell migration.

Effect of low frequency ultrasound waves on the morphology and viability of cultured human gingival fibroblasts

ObjectivesThe aim of this study was to investigate the effect of the vibration amplitude of mechanical ultrasound waves (27 kHz) on the viability and morphology of human gingival fibroblasts (hGFs) when cultured on a biomaterial substrate. MethodhGFs were seeded on tissue culture plates (TCPs) and an Ti6Al4V titanium alloy surface in two groups for three days and seven days of cell culture. The cells were subjected to three vibration amplitudes for 20 min each day. Scanning electron microscope (SEM) images were used to characterize cell morphology. ResultsExperiments showed that hGF cells became detached from their plates at a vibration amplitude comparable to an intensity of 260 mW/cm2. In addition, hGfs that received a vibrational amplitude comparable to an intensity of 50 mW/cm2 underwent significant proliferation proliferated significantly; however, cells receiving higher amplitudes suffered from adverse effects. ConclusionsSEM images of hGFs on titanium disks at vibration amplitude comparable to an intensity 50 mW/cm2 showed a remarkable hexagonal architecture, which we refer to as a honeycomb pattern. On day 6 the observed hGFs on TCPs, proliferated at a higher rate and new cells attached uniformly on the existing layer of cells. These data indicate the effect of cellular tissue as a substrate on the growth of new hGFs under low-intensity ultrasound.

Multi-Omics Data Integration Reveals Key Variables Contributing to Subgingival Microbiome Dysbiosis-Induced Inflammatory Response in a Hyperglycemic Microenvironment.

Subgingival microbiome dysbiosis promotes the development of periodontitis, an irreversible chronic inflammatory disease associated with metabolic diseases. However, studies regarding the effects of a hyperglycemic microenvironment on host-microbiome interactions and host inflammatory response during periodontitis are still scarce. Here, we investigated the impacts of a hyperglycemic microenvironment on the inflammatory response and transcriptome of a gingival coculture model stimulated with dysbiotic subgingival microbiomes. HGF-1 cells overlaid with U937 macrophage-like cells were stimulated with subgingival microbiomes collected from four healthy donors and four patients with periodontitis. Pro-inflammatory cytokines and matrix metalloproteinases were measured while the coculture RNA was submitted to a microarray analysis. Subgingival microbiomes were submitted to 16s rRNA gene sequencing. Data were analyzed using an advanced multi-omics bioinformatic data integration model. Our results show that the genes krt76, krt27, pnma5, mansc4, rab41, thoc6, tm6sf2, and znf506 as well as the pro-inflammatory cytokines IL-1β, GM-CSF, FGF2, IL-10, the metalloproteinases MMP3 and MMP8, and bacteria from the ASV 105, ASV 211, ASV 299, Prevotella, Campylobacter and Fretibacterium genera are key intercorrelated variables contributing to periodontitis-induced inflammatory response in a hyperglycemic microenvironment. In conclusion, our multi-omics integration analysis unveiled the complex interrelationships involved in the regulation of periodontal inflammation in response to a hyperglycemic microenvironment.

Novel Herbal Therapeutic YH23537 Improves Clinical Parameters in Ligature-Induced Periodontal Disease Model in Beagle Dogs.

Currently, available medicine does not satisfy the clinical unmet needs of periodontal disease. Therefore, novel drugs with improved efficacy profiles are needed. We previously demonstrated that YH14642, water extracts of Notoginseng Radix and Rehmanniae Radix Preparata, improved probing depths in double-blind phase II clinical trial. However, it still has hurdles for commercialization due to the low efficiency of active compound extraction. To resolve this issue, we developed YH23537 through process optimization to extract active compounds efficiently while still achieving the chemical profile of YH14642. In this study, we investigated the therapeutic effects of YH23537 compared with YH14642 using a canine model of ligature-induced periodontitis. Human gingival fibroblast (hGF) cells were treated with various concentrations of YH23537 or YH14642 with lipopolysaccharide (LPS) for 24 hr. IL-6 and IL-8 levels in the conditioned media were determined using Luminex. Sixteen 3-year-old male beagle dogs had their teeth scaled and polished using a piezo-type ultrasonic scaler under general anesthesia and brushed once daily for the following 2 weeks. Two weeks after the scaling procedure, the left upper second premolar (PM2), third premolar (PM3), and fourth premolar (PM4) as well as the left lower PM3, PM4, and first molar (M1) were ligated with silk-wire twisted ligatures. The dogs were fed with soft moistened food to induce periodontitis for 8 weeks, and the ligatures were then removed. YH23537 and YH14642 were administered for 4 weeks, and clinical periodontal parameters such as plaque index (PI), gingival index (GI), probing depth (PD), clinical attachment level (CAL), and bleeding on probing (BoP) were determined before and 1, 2, 3, and 4 weeks after treatment. YH23537 inhibited IL-6 and IL-8 secretion in a dose-dependent manner in hGF cells stimulated with LPS. The IC50 values for YH23537 were 43 and 54 μg/ml for IL-6 and IL-8, respectively, while the values for YH14642 were 104 and 117 μg/ml, respectively. In the animal study, clinical parameters including GI, PD, CAL, and BoP were significantly increased after 8 weeks of ligature-induced periodontitis. The YH23537 300 and YH23537 900 mg groups had significant improvements in CAL from 1 to 4 weeks after treatment in comparison to the placebo group. GR values in the YH23537 900 mg group were decreased throughout the treatment period. GI values were also reduced significantly after 4-week treatment with 300 and 900 mg of YH23537. YH23537 at 300 mg doses showed comparable efficacy for CAL and GR with 1,000 mg of YH14642. YH23537 showed therapeutic efficacy against periodontitis in dogs, mediated by anti-inflammatory effects. These findings indicate that YH23537 has the potential for further development as a new drug for patients suffering from periodontal disease.

S-1-propenyl-L-cysteine suppresses lipopolysaccharide-induced expression of matrix metalloproteinase-1 through inhibition of tumor necrosis factor-α converting enzyme-epidermal growth factor receptor axis in human gingival fibroblasts.

Periodontal disease is the most common dental health problem characterized by the destruction of connective tissue and the resorption of alveolar bone resulting from a chronic infection associated with pathogenic bacteria in the gingiva. Aged garlic extract has been reported to improve gingival bleeding index and probing pocket depth score in patients with mild to moderate periodontitis. Although our previous study found that aged garlic extract and its constituents suppressed the tumor necrosis factor-α-induced inflammatory responses in a human gingival epithelial cell line, the mechanism underlying the effect of aged garlic extract on the destruction of the gingiva remains unclear. The present study investigated the effect of S-1-propenyl-L-cysteine, one of the major sulfur bioactive compounds in aged garlic extract, on the lipopolysaccharide-induced expression of matrix metalloproteinases in human gingival fibroblasts HGF-1 cells. Matrix metalloproteinases are well known to be closely related to the destruction of the gingiva. We found that S-1-propenyl-L-cysteine suppressed the lipopolysaccharide-induced expression and secretion of matrix metalloproteinase-1 in HGF-1 cells. In addition, S-1-propenyl-L-cysteine inhibited the lipopolysaccharide-induced phosphorylation of epidermal growth factor receptor and expression of the active form of tumor necrosis factor-α converting enzyme. Furthermore, the inhibitors of epidermal growth factor receptor tyrosine kinase and tumor necrosis factor-α converting enzyme, AG-1478 and TAPI-1, respectively, reduced the lipopolysaccharide-induced protein level of matrix metalloproteinase-1, as did S-1-propenyl-L-cysteine. Taken together, these results suggested that S-1-propenyl-L-cysteine suppresses the lipopolysaccharide-induced expression of matrix metalloproteinase-1 through the blockade of the tumor necrosis factor-α converting enzyme-epidermal growth factor receptor axis in gingival fibroblasts.

Human Gingival Fibroblasts as a Novel Cell Model Describing the Association between Bitter Taste Thresholds and Interleukin-6 Release

Human gingival fibroblast cells (HGF-1 cells) present an important cell model to investigate the gingiva's response to inflammatory stimuli such as lipopolysaccharides from Porphyromonas gingivalis (Pg-LPS). Recently, we demonstrated trans-resveratrol to repress the Pg-LPS evoked release of the pro-inflammatory cytokine interleukin-6 (IL-6) via involvement of bitter taste sensing receptor TAS2R50 in HGF-1 cells. Since HGF-1 cells express most of the known 25 TAS2Rs, we hypothesized an association between a compound's bitter taste threshold and its repressing effect on the Pg-LPS evoked IL-6 release by HGF-1 cells. To verify our hypothesis, 11 compounds were selected from the chemical bitter space and subjected to the HGF-1 cell assay, spanning a concentration range between 0.1 μM and 50 mM. In the first set of experiments, the specific role of TAS2R50 was excluded by results from structurally diverse TAS2R agonists and antagonists and by means of a molecular docking approach. In the second set of experiments, the HGF-1 cell response was used to establish a linear association between a compound's effective concentration to repress the Pg-LPS evoked IL-6 release by 25% and its bitter taste threshold concentration published in the literature. The Pearson correlation coefficient revealed for this linear association was R2 = 0.60 (p < 0.01), exceeding respective data for the test compounds from a well-established native cell model, the HGT-1 cells, with R2 = 0.153 (p = 0.263). In conclusion, we provide a predictive model for bitter tasting compounds with a potential to act as anti-inflammatory substances.

Leaching components and initial biocompatibility of novel bioactive restorative materials.

Bioactive restorative materials were developed on the premise that direct restorations should not only serve the purpose of reconstructing dental hard tissue defects but also exhibit biological features that prevent secondary caries development, without having adverse effects on the host cells. This study focuses on assessing the in vitro biocompatibility of two novel bioactive restorative materials. Specimens of the bioactive restorative materials, Cention Forte (CF) and ACTIVA BioACTIVE RESTORATIVE (AB), a glass ionomer cement/glass hybrid (EQUIA Forte HT, EF) and an established nanohybrid composite (Venus Diamond, VD) were produced and finished. The specimens were eluted in water and methanol and the resulting eluates were analyzed via gas chromatography-mass spectrometry. hGF-1 cells were exposed to eluates prepared in cell culture medium. Cellular ATP levels, oxidized glutathione concentration, caspase-3/7 activity and the inflammatory response (IL-6 and PGE2 levels) were determined. Microscopic images were taken to examine the cell morphology. Methyl methacrylate and 2-Hydroxyethyl methacrylate were the main monomers detected in CF and AB eluates. All materials inhibited cell proliferation and led to significantly reduced ATP-levels. The cells exhibited a healthy morphology in the presence of CF and AB. Cells exposed to VD showed increased oxidized glutathione levels. Only EF led to enhanced caspase-3/7 activity. CF and AB caused IL-6 levels to increase, while EF and AB led to enhanced PGE2 levels. CF and AB are promising materials from a biological point of view and seem to have improved bioactive properties compared to glass ionomer cements.

Chemo-radiation therapy of U87-MG glioblastoma cells using SPIO@AuNP-Cisplatin-Alginate nanocomplex

Megavoltage radiotherapy and cisplatin-based chemotherapy are the primary glioblastoma treatments. Novel nanoparticles have been designed to reduce adverse effects and boost therapeutic effectiveness. In the present study, we synthesized the SPIO@AuNP-Cisplatin-Alginate (SACA) nanocomplex, composed of a SPIO core, a gold shell, and an alginate coating. SACA was characterized using transmission electron microscopy (TEM) and dynamic light scattering (DLS). U87-MG human glioblastoma cells and the HGF cell line (a healthy primary gingival fibroblast) were treated in multiple groups by a combination of SACA, cisplatin, and 6 MV X-ray. The MTT assay was used to assess the cytotoxicity of cisplatin and SACA (at various concentrations and for 4 h). Following the treatments, apoptosis and cell viability were evaluated in each treatment group using flow cytometry and the MTT assay, respectively. The findings demonstrated that the combination of SACA and 6 MV X-rays (at the doses of 2 and 4 Gy) drastically decreased the viability of U87MG cells, whereas the viability of HGF cells remained unchanged. Moreover, U87MG cells treated with SACA in combination with radiation exhibited a significant increase in apoptosis, demonstrating that this nanocomplex effectively boosted the radiosensitivity of cancer cells. Even though additional in vivo studies are needed, these findings suggest that SACA might be used as a radiosensitizer nanoparticle in the therapy of brain tumors.

Osteoblasts and Fibroblasts Interaction with a Porcine Acellular Dermal Matrix Membrane.

The use of collagen membranes has remained the gold standard in GTR/GBR. In this study, the features and the biological activities of an acellular porcine dermis collagen matrix membrane applicable during dental surgery were investigated, and also by applying hydration with NaCl. Thus, two tested membranes were distinguished, the H-Membrane and Membrane, compared to the control cell culture plastic. The characterization was performed by SEM and histological analyses. In contrast, the biocompatibility was investigated on HGF and HOB cells at 3, 7, and 14 days by MTT for proliferation study; by SEM and histology for cell interaction study; and by RT-PCR for function-related genes study. In HOBs seeded on membranes, mineralization functions by ALP assay and Alizarin Red staining were also investigated. Results indicated that the tested membranes, especially when hydrated, can promote the proliferation and attachment of cells at each time. Furthermore, membranes significantly increased ALP and mineralization activities in HOBs as well as the osteoblastic-related genes ALP and OCN. Similarly, membranes significantly increased ECM-related and MMP8 gene expression in HGFs. In conclusion, the tested acellular porcine dermis collagen matrix membrane, mainly when it is hydrated, behaved as a suitable microenvironment for oral cells.

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.

Impact of Metal Particles Released during Ultrasonic Scaling of Titanium Surfaces on Human Gingival Fibroblasts.

Metal particles found in tissues around dental implants have been proposed to play a pathogenic role in peri-implantitis. Ultrasonic scaling has been suggested as a mechanism by which these particles can be inadvertently released into surrounding tissues. Furthermore, risk factors like diabetes can result in exacerbation of this inflammatory condition. The current study aimed to analyze metal particles released from titanium surfaces during ultrasonic scaling and their impact on pro-inflammatory cytokine production by human gingival fibroblasts. Metal particles generated from ultrasonic scaling of titanium discs using two different tips (metal and poly-etheretherketone tips) were characterized using scanning electron microscopy and elemental analysis. Endotoxin levels and Human gingival fibroblast viability, in the presence commercial and ultrasonically generated particles were determined. Fibroblasts, cultured in high or low glucose growth medium, were incubated with commercial titanium particles or ultrasonically generated particles in the presence or absence of interluekin-1β. Interleukin 6 and interleukin 8 production were then quantified using Enzyme linked immunosorbent assay. Analysis of particles after scaling of titanium discs showed significant levels of titanium particles. Commercial titanium particles and generated particles had no effect of fibroblast viability. Endotoxin levels of all particles were too low to stimulate HGF cells. IL-1β significantly stimulated IL-6 and IL-8 production. However, commercial, and generated particles generally had no significant effect on IL- 6 and IL-8 production. Our study concluded that particles generated during ultrasonic scaling had no significant effect on viability of HGF cells and cytokine production.

Eugenol Induces Apoptosis in Tongue Squamous Carcinoma Cells by Mediating the Expression of Bcl-2 Family.

Head and neck squamous cell carcinoma is highly aggressive type of cancer for which the available treatment often causes patients severe side effects. Eugenol (Eug) is the major active constituent of clove essential oil and is known to possess antitumor properties. The present study aimed to assess the in vitro cytotoxicity of eugenol in SCC-4, tongue squamous carcinoma cells, and also in HGF, human gingival fibroblasts. Both cell lines were treated with five concentrations of Eug (0.1-1 mM) for 72 h. Cellular viability was assessed, followed by cellular morphological evaluation and by staining of the nuclei and cytoskeleton. RT-PCR was conducted in order to find the effect eugenol had on the expression on Bad, Bax, and Bcl-2 genes. Eugenol induced a dose-dependent decrease in viability in both cell lines, with the SCC-4 cells being significantly more affected. HGF cells detached from the plate at the highest concentrations used, while SCC-4 cells changed their morphology in a dose-dependent manner, with rounding, floating cells, and confluency loss being observed. Apoptotic-like signs such as chromatin and actin filaments condensation were clearly seen in SCC-4 cells, while RT-PCR revealed a significantly increased expression of pro-apoptotic genes Bax and Bad. Therefore, eugenol exerts its cytotoxic effect in tongue squamous cell carcinoma through inducing apoptosis.

Synbiotic-fluoride synergism on enamel remineralization, cytotoxicity and genotoxicity

Objective(s)The objectives of the present study were to examine the – a) enamel remineralization potential of synbiotic-fluoride (SF) therapy using a multi-species bacterial pH-cycling model; and b) cytotoxic and genotoxic effects of SF therapy extracts. Materials and methodsThe SF therapy group comprised of 2% arginine (Arg), 0.2% NaF, and a probiotic Lactobacillus rhamnosus GG (LRG). The intervention groups studied were: 1) No treatment; 2) 2% Arg; 3) 0.2% NaF; 4) LRG; 5) 2% Arg+0.2% NaF; 6) 2% Arg+LRG; 7) 0.2% NaF+LRG; and 8) 2% Arg+0.2% NaF+LRG (SF therapy). The enamel remineralization potential of SF therapy was investigated under cariogenic biofilm challenge; while the cytotoxicity and genotoxicity of SF therapy extracts were examined on HGF-1 and Chinese hamster fibroblast V79, respectively. To determine the remineralization effect, the specimens were subjected to mineral density (MD) assessment using micro-CT, Ca/P molar ratio with SEM-EDX, and enamel fluoride uptake (EFU) estimates. The HGF-1 proliferation assessment was quantified using MTT/CCK-8 assays with qualitative analysis by nuclei staining Hoechst-based fluorescence imaging. The genotoxicity was determined by micronuclei formation test. ResultsMineral gain and %remineralization derived from MD assessment for the SF therapy were significantly higher than the other groups (p<0.05). The %ΔCa/P for the SF and 2% Arg+0.2% NaF were significantly higher than the other groups (p<0.05). The SF and 2% Arg+0.2% NaF groups had the highest EFU compared to the other groups (p<0.05). No significant difference in the %viable HGF-1 cells were observed between the treatment interventions and no treatment group (p>0.05). Compared to the EMS-positive control, the micronuclei formation for all the intervention groups was significantly lower (p<0.05), with no significant difference among the treatment groups (p>0.05). ConclusionThe SF therapy enhanced enamel remineralization with no biocompatibility concerns. Clinical significanceWith the enhanced enamel remineralization potential discerned in the present study, the SF therapy can be used as a promising caries-preventive agent targeted for high caries-risk individuals.

1-[4-(2-Dimethylaminoethoxy)phenylcarbonyl]-3,5-Bis(3,4,5-Trimethoxybenzylidene)- 4-Piperidone Hydrochloride and Related Compounds: Potent Cytotoxins Demonstrate Greater Toxicity to Neoplasms than Non- Malignant Cells.

The incidence of cancer has been increasing worldwide. Unfortunately, the drugs used in cancer chemotherapy are toxic to both neoplasms and normal tissues, while many available medications have low potencies. Conjugated α,β-unsaturated ketones differ structurally from contemporary anticancer medications , some of which have noteworthy antineoplastic properties. This study aimed to design and synthesize highly potent cytotoxins with far greater toxicity to neoplasms than to non-malignant cells. A series of N-acyl-3,5-bis(benzylidene)-4-piperidone hydrochlorides 4a-n were prepared and evaluated against Ca9-22, HSC-2, HSC-3, and HSC-4 squamous cell carcinomas as well as against HGF, HPLF, and HPC non-malignant cells. QSAR and western blot analyses were performed. The majority of compounds display submicromolar CC50 values towards the neoplasms; the figures for some of the compounds are below 10-7 M. In general, 4a-n have much lower CC50 values than those of melphalan, 5-fluorouracil, and methotrexate, while some compounds are equitoxic with doxorubicin. The compounds are far less toxic to the non-malignant cells, giving rise to substantial selectivity index (SI) figures. A QSAR study revealed that both potency and the SI data were controlled to a large extent by the electronic properties of the substituents in the arylidene aryl rings. Two representative compounds, 4f and 4g, caused apoptosis in HSC-2 cells. The compounds in series 4 are potent cytotoxins displaying tumor-selective toxicity. In particular, 4g with an average CC50 value of 0.04 μM towards four malignant cell lines and a selectivity index of 46.3 is clearly a lead molecule that should be further evaluated.