HGF Cell Research Articles

Immunomodulatory, Antioxidant, and Potential Anticancer Activity of the Polysaccharides of the Fungus Fomitiporia chilensis.

Fomitiporia species have aroused the interest of numerous investigations that reveal their biological activity and medicinal potential. The present investigation shows the antioxidant, anticancer, and immunomodulatory activity of acidic polysaccharides obtained from the fungus Fomitiporia chilensis. The acidic polysaccharides were obtained for acidic precipitation with 2% O-N-cetylpyridinium bromide. Chemical analysis was performed using FT-IR and GC-MS methods. The antioxidant capacity of acidic polysaccharides from F. chilensis was evaluated by scavenging free radicals with an ABTS assay. Macrophage proliferation and cytokine production assays were used to determine the immunomodulatory capacity of the polysaccharides. Anti-tumor and cytotoxicity activity was evaluated with an MTT assay in the U-937, HTC-116, and HGF-1 cell lines. The effect of polysaccharides on the cell cycle of the HCT-116 cell line was determined for flow cytometry. Fourier Transform-infrared characterization revealed characteristic absorption peaks for polysaccharides, whereas the GC-MS analysis detected three peaks corresponding to D-galactose, galacturonic acid, and D-glucose. The secreted TNF-α concentration was increased when the cell was treated with 2 mg mL-1 polysaccharides, whereas the IL-6 concentration was increased with all of the evaluated polysaccharide concentrations. A cell cycle analysis of HTC-116 treated with polysaccharides evidenced that the acidic polysaccharides from F. chilensis induce an increase in the G0/G1 cell cycle phase, increasing the apoptotic cell percentage. Results from a proteomic analysis suggest that some of the molecular mechanisms involved in their antioxidant and cellular detoxifying effects and justify their traditional use in heart diseases. Proteomic data are available through ProteomeXchange under identifier PXD048361. The study on acidic polysaccharides from F. chilensis has unveiled their diverse biological activities, including antioxidant, anticancer, and immunomodulatory effects. These findings underscore the promising therapeutic applications of acidic polysaccharides from F. chilensis, warranting further pharmaceutical and medicinal research exploration.

Anticandidal effect of cinnamic acid characterized from Cinnamomum cassia bark against the fluconazole resistant strains of Candida.

Candida spp., causes invasive fungal infections, especially in immune-compromised patients and the propensity of antifungal resistance against azole-based drugs need to be addressed. This study is thus aimed to characterize the anticandidal effect of the cinnamic acid extracted from the barks of Cinnamomum cassia. Five species of Fluconazole-resistant Candida sp. were retrieved from the department repertoire. The extraction of CA was performed by three different methods followed by silica gel column chromatography. Eluant was subjected to FTIR and XRD analysis for confirmation. The anticandidal activity of the CA was checked by the agar disc diffusion method and the MIC and MFC were determined. The anti-biofilm effect of CA was assessed using the CLSM technique followed by the biocompatibility check using MTT assay in normal HGF cell lines. CA was best extracted with the hot maceration method using ethanol with a maximum yield of 6.73mg. Purification by column chromatography was achieved using benzene, acetic acid, and water (6:7:3) mobile phase. CA was confirmed by FTIR with absorption peaks and by XDR based on strong intensity. CA was found to possess promising anticandidal activity at 8µg/mL with MIC and MFC values determined as 0.8µg/mL and 0.08µg/mL respectively. Antibiofilm activity by CLSM analysis revealed biofilm inhibition and was biocompatible at 8.5µg/ml concentrations in HGF cell lines until 24h. The study findings conclude that CA is the best alternative to treat candidal infection warranting further experimental preclinical studies.

Development of Novel Antibacterial Ti-Nb-Ga Alloys with Low Stiffness for Medical Implant Applications.

With the rising demand for medical implants and the dominance of implant-associated failures including infections, extensive research has been prompted into the development of novel biomaterials that can offer desirable characteristics. This study develops and evaluates new titanium-based alloys containing gallium additions with the aim of offering beneficial antibacterial properties while having a reduced stiffness level to minimise the effect of stress shielding when in contact with bone. The focus is on the microstructure, mechanical properties, antimicrobial activity, and cytocompatibility to inform the suitability of the designed alloys as biometals. Novel Ti-33Nb-xGa alloys (x = 3, 5 wt%) were produced via casting followed by homogenisation treatment, where all results were compared to the currently employed alloy Ti-6Al-4V. Optical microscopy, scanning electron microscopy (SEM), and energy dispersive spectroscopy (EDS) results depicted a single beta (β) phase microstructure in both Ga-containing alloys, where Ti-33Nb-5Ga was also dominated by dendritic alpha (α) phase grains in a β-phase matrix. EDS analysis indicated that the α-phase dendrites in Ti-33Nb-5Ga were enriched with titanium, while the β-phase was richer in niobium and gallium elements. Mechanical properties were measured using nanoindentation and microhardness methods, where the Young's modulus for Ti-33Nb-3Ga and Ti-33Nb-5Ga was found to be 75.4 ± 2.4 and 67.2 ± 1.6 GPa, respectively, a significant reduction of 37% and 44% with respect to Ti-6Al-4V. This reduction helps address the disproportionate Young's modulus between titanium implant components and cortical bone. Importantly, both alloys successfully achieved superior antimicrobial properties against Gram-negative P. aeruginosa and Gram-positive S. aureus bacteria. Antibacterial efficacy was noted at up to 90 ± 5% for the 3 wt% alloy and 95 ± 3% for the 5 wt% alloy. These findings signify a substantial enhancement of the antimicrobial performance when compared to Ti-6Al-4V which exhibited very small rates (up to 6.3 ± 1.5%). No cytotoxicity was observed in hGF cell lines over 24 h. Cell morphology and cytoskeleton distribution appeared to depict typical morphology with a prominent nucleus, elongated fibroblastic spindle-shaped morphology, and F-actin filamentous stress fibres in a well-defined structure of parallel bundles along the cellular axis. The developed alloys in this work have shown very promising results and are suggested to be further examined towards the use of orthopaedic implant components.

Antibacterial and cytotoxic effects of fresh bovine amniotic membrane with hydroxyapatite (BAM-HA): a laboratory experiment

Introduction: Bacterial infections, particularly by Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans) and Porphyromonas gingivalis (P. gingivalis), can worsen alveolar bone resorption after tooth extraction. The capability of Bovine Amniotic Membrane-Hydroxyapatite (BAM-HA) biocomposite to reduce this resorption has been explored. However, before clinical use, cytotoxicity testing is imperative to ensure its biocompatibility. The aim of the study was to assess both the antibacterial effects and cytotoxicity of the BAM-HA biocomposite to ensure its suitability for clinical use biocompatibility of the BAM-HA biocomposite before its clinical application. Methods: The laboratory-based research involved testing BAM combined with HA powder in 4:1 and 4:2 ratios via freeze-drying and underwent antibacterial tests against A. actinomycetemcomitans and P. gingivalis, using the plate count method. Cytotoxicity tests were performed on HGF cells, including negative control, positive control, BAM-HA (4:1), and BAM-HA (4:2) groups, with statistical analysis conducted using One-Way ANOVA and Post Hoc Bonferroni and Tukey tests. Results: Antibacterial tests against A. actinomycetemcomitans revealed significant reduction in colony count with BAM-HA ratios 4:1 (129.0 ± 12.7 CFU/mL) and 4:2 (77.3 ± 15.5 CFU/mL) compared to the negative control (186.6 ± 27.5 CFU/mL). Similar reductions were observed for P. gingivalis, with BAM-HA ratios 4:1 (51.3 ± 6.6 CFU/mL) and 4:2 (3.1 ± 1.5 CFU/mL) compared to the negative control (117.3 ± 22.0 CFU/mL). Cytotoxicity tests showed no significant differences in HGF cell viability and IC50 values between the negative control and BAM-HA (4:1) or BAM-HA (4:2) groups. Conclusion: The BAM - HA biocomposite shows antibacterial effects against A. actinomycetemcomitans and P. gingivalis. Moreover, BAM - HA ratios of 4:1 and 4:2 do not induce cytotoxic effects on human gingival fibroblasts, suggesting potential biocompatibility for clinical applications.KeywordA. actinomycetemcomitans, antibacterial effects, BAM-HA biocomposite, cytotoxicity, P. gingivalis

Bifunctional folic acid targeted biopolymer Ag@NMOF nanocomposite [{Zn2 (1,4-bdc) 2 (DABCO)} n] as a novel theranostic agent for molecular imaging of colon cancer by SERS

Without a doubt, cancer and its negative impact on human health have created many hurdles for people across the world since conventional approaches have not offered a reliable ability in the eradication of cancer. As a result, finding novel approaches, like using bimodal nanoparticles as a potential nanocarrier in molecular imaging and cancer therapy, is remarkably required these days. In the present study, ex-situ (Ge) and in-situ (Gi) green synthesized silver (Ag) nanoparticles entrapped in metal-organic framework nanocomposites (NMOF) coated with folic acid (FA) targeted chitosan (CS) was successfully developed as a novel bifunctional nanocarrier for detection and treatment of colon cancer cells. Then nanocarriers, such as NMOF–CS–FA, Ge–Ag@NMOF–CS–FA, Gi-Ag@NMOF–CS–FA, and C–Ag@NMOF–CS–FA, were characterized via FT-IR, DLS, SERS, TEM, and SEM and results have potentially confirmed the quality and quantity of synthesized nanocomposites. The hydrodynamic diameters of NMOF-CS, Ge–Ag@NMOF-CS, Gi-Ag@NMOF-CS, and C–Ag@NMOF-CS specimens were measured at around 99.7 ± 10 nm, 110 ± 10 nm, 118 ± 10 nm, 115 ± 10 nm, respectively. Also, the PDI values less than 0.2 confirm the reliable distribution of these nanocomposites. Afterward, the cell viability assay was conducted on HCT116 and HGF cell lines for evaluating biocompatibility and targeting efficiency of nanocomposites; FA functionalized nanocomposites have intensively indicated better performance in cancer cells targeting and their inhibition, and IC50 was attained for 10 ng/mL of Ge–Ag@NMOF–CS–FA while non-targeted nanocarriers did not have toxicity more than 20 % on HCT116 colon cancer cells. Moreover, according to the results, the cell viability of HGF normal cells was at least 85 % after being exposed to different concentrations of nanocomposites for 24 h. This indicates that the synthesized nanocomposites do not have significant toxic effects on normal cells. The results indicate that this novel nanocomposite has the potential to effectively deliver drugs to cancer cells.

Genoprotective Potency of Vit C on Zinc Oxide (ZnO) and Iron Oxide (Fe2O3) Nanoparticle-Mediated Toxicity, an In Vitro Study

Introduction: The potential of zinc oxide (ZnO) and iron oxide (Fe2O[Formula: see text] nanoparticles (NPs) to induce toxic effects, especially genotoxicity, has been demonstrated in previous studies and is in part related to the ability of NPs to produce ROS. The use of antioxidants is an effective method to reduce NP-induced genotoxicity. The aim of this study was to determine the protective role of vitamin C as a potent antioxidant in ZnO-and Fe2O3 NP-induced genotoxicity in the HGF-1 cell line. Methods: Different concentrations of ZnO and Fe2O3 NPs (50[Formula: see text][Formula: see text]g, 100[Formula: see text][Formula: see text]g, and 150[Formula: see text][Formula: see text]g/mL) were used to achieve the best concentration for further evaluation. HGF-1 cells were incubated with different concentrations of vitamin C 24[Formula: see text]h before the NPs. The cells were then exposed to ZnO and Fe2O3 NPs at a concentration of 100[Formula: see text][Formula: see text]g/mL for 1[Formula: see text]h. The possible genoprotective effects of vitamin C were determined using a comet assay. Results: The results of this study showed that all concentrations of vitamin C could reduce the DNA damage induced by ZnO and Fe2O3 NPs. Discussion: In conclusion, vitamin C could be considered a potent genoprotective agent against ZnO- and Fe2O3 NP-induced genotoxicity.

The Phenolic Profile and Anti-Inflammatory Effect of Ethanolic Extract of Polish Propolis on Activated Human Gingival Fibroblasts-1 Cell Line.

Propolis, owing to its antibacterial and anti-inflammatory properties, acts as a cariostatic agent, capable of preventing the accumulation of dental plaque and inhibiting inflammation. The anti-inflammatory properties of propolis are attributed to caffeic acid phenethyl ester (CAPE), which is present in European propolis. The objective of the conducted study was to assess the anti-inflammatory effects of the Polish ethanolic extract of propolis (EEP) and isolated CAPE on stimulated with LPS and IFN-α, as well as the combination of LPS and IFN-α. The cytotoxicity of the tested compounds was determined using the MTT assay. The concentrations of specific cytokines released by the HGF-1 cell line following treatment with EEP (25-50 µg/mL) or CAPE (25-50 µg/mL) were assessed in the culture supernatant. In the tested concentrations, both CAPE and EEP did not exert cytotoxic effects. Our results demonstrate that CAPE reduces TNF-α and IL-6 in contrast to EEP. Propolis seems effective in stimulating HGF-1 to release IL-6 and IL-8. A statistically significant difference was observed for IL-8 in HGF-1 stimulated by LPS+IFN-α and treated EEP at a concentration of 50 µg/mL (p = 0.021201). Moreover, we observed that CAPE demonstrates a stronger interaction with IL-8 compared to EEP, especially when CAPE was administered at a concentration of 50 µg/mL after LPS + IFN-α stimulation (p = 0.0005). Analysis of the phenolic profile performed by high-performance liquid chromatography allowed identification and quantification in the EEP sample of six phenolic acids, five flavonoids, and one aromatic ester-CAPE. Propolis and its compound-CAPE-exhibit immunomodulatory properties that influence the inflammatory process. Further studies may contribute to explaining the immunomodulatory action of EEP and CAPE and bring comprehensive conclusions.

Safety Assessment and Hepatic-Renal Protection of Cajanus cajan (L.) Millsp. Root and Its Soy Isoflavone Contents.

Cajanus cajan (L.) Millsp., also known as pigeon pea, has roots that have exhibited much pharmacological potential. The present study was conducted to assess the safe dose of the ethanolic extract of C. cajan roots (EECR95) and to analyze the main soy isoflavones contents. In vitro, we investigated the mutagenicity and cytotoxic effect of EECR95 on Salmonella typhimurium-TA98 and TA100 (by Ames tests) and RAW 264.7, L-929, and HGF-1 cell lines (by MTT tests) for 24 h of incubation. We found no mutagenic or cytotoxic effects of EECR95. After administration of 0.2 or 1.0 g/kg bw of EECR95 to both male and female Wistar rats for 90 days, there were no significant adverse effects on the behaviors (body weight, water intake, and food intake), organ/tissue weights, or immunohistochemical staining, and the urine and hematological examinations of the rats were within normal ranges. EECR95 potentially decreases renal function markers in serum (serum uric acid, BUN, CRE, and GLU) or liver function markers (cholesterol, triglyceride, and glutamic-pyruvate-transaminase (GPT)). We also found that EECR95 contained five soy isoflavones (genistein, biochanin A, daidzein, genistin, and cajanol), which may be related to its hepatorenal protection. Based on the high dose (1.0 g/kg bw) of EECR95, a safe daily intake of EECR95 for human adults is estimated to be 972 mg/60 kg person/day.

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.

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.

An organotypic oral mucosal infection model to study host-pathogen interactions

Early in vitro oral mucosal infection models (OMMs) failed to consider the suitability of the model environment to represent the host immune response. Denture stomatitis (DS) is mediated by Candida albicans, but the role of Staphylococcus aureus remains uncertain. A collagen hydrogel-based OMM containing HaCaT and HGF cell types was developed, characterised and employed to study of tissue invasion and pro-inflammatory cytokine production in response to pathogens. Models formed a robust epithelium. Despite their inflammatory baseline, 24-h infection with C. albicans, and/or S. aureus led to tissue invasion, and significantly upregulated IL-6 and IL-8 production by OMMs when compared to the unstimulated control. No significant difference in IL-6 or IL-8 production by OMMs was observed between single and dual infections. These attributes indicate that this newly developed OMM is suitable for the study of DS and could be implemented for the wider study of oral infection.

Utilization of a nanostructured lipid carrier encapsulating pitavastatin–Pinus densiflora oil for enhancing cytotoxicity against the gingival carcinoma HGF-1 cell line

Oral squamous cell carcinoma (OSCC) is the most common epithelial tumor of the oral cavity. Gingival tumors, a unique type of OSCC, account for 10% of these malignant tumors. The antineoplastic properties of statins, including pitavastatin (PV), and the essential oil of the Pinus densiflora leaf (Pd oil) have been adequately reported. The goal of this investigation was to develop nanostructured lipid carriers (NLCs) containing PV combined with Pd oil and to determine their cytotoxicity against the cell line of human gingival fibroblasts (HGF-1). A central composite quadratic design was adopted to optimize the nanocarriers. The particle size and stability index of the nano-formulations were measured to evaluate various characteristics. TEM analysis, the entrapment efficiency, dissolution efficiency, and the cytotoxic efficiency of the optimized PV-loaded nanostructured lipid carrier drug delivery system (PV-Pd-NLCs) were evaluated. Then, the optimal PV-Pd-NLCs was incorporated into a Carbopol 940® gel base and tested for its rheological features and its properties of release and cell viability. The optimized NLCs had a particle size of 98 nm and a stability index of 89%. The gel containing optimum PV-Pd-NLCs had reasonable dissolution efficiency and acceptable rheological behavior and acquired the best cytotoxic activity against HGF-1 cell line among all the formulations developed for the study. The in vitro cell viability studies revealed a synergistic effect between PV and Pd oil in the treatment of gingival cancer. These findings illustrated that the gel containing PV-Pd-NLCs could be beneficial in the local treatment of gingival cancer.

Preparation and In-Vitro Evaluation of Ketoconazole-Loaded Niosome (Ketosome) for Drug Delivery to Cutaneous Candidiasis

Background: Recently, niosomes are becoming popular in drug delivery. The current work aimed to investigate the characteristics, cellular safety, and antifungal activity of ketoconazole-loaded niosome (ketosome). Methods: Ultrasonic approach was employed to prepare ketosome including cholesterol, nonionic surfactant and ketoconazole. The size characteristics and morphological features of ketosome and physicochemical properties of ketoconazole in ketosomes were evaluated using dynamic light scattering (DLS), differential scanning calorimetry (DSC), powder x-ray diffractometer (PXRD), scanning electron microscopy (SEM), and attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy. Also, the dissolution rate, cellular safety test and antimycotic properties of ketosome were examined. Results: According to the results, the particle size of the ketosome decreased from 491.400±10.622 to 121.300±7.274 nm by the increment of cholesterol. According to further research, changes in the cholesterol:surfactants ratio can modulate the zeta potential from -27.866±1.069 to -12.500±1.153 mV. The highest entrapment of ketoconazole was about 87% when the cholesterol concentration in the ketosome was high. Ketosome with the maximum cholesterol:surfactants ratio showed the fastest drug release. Furthermore, the cell viability assay revealed that the ketosome had lower cytotoxicity in comparison with pure drug. The cell viability of the ketosome was estimated to be about 90% (HGF cell line). The ketosome had a lower MIC than the pure drug when tested against Candida albicans. Conclusion: The results of this study revealed that the optimized ketoconazole-loaded niosome could be used as a possible nanovesicle for ketoconazole drug delivery, potentially opening up new ways for the management of cutaneous candidiasis complaints.

Therapeutic Effect of Green Synthesized Silver Nanoparticles Using Erodium glaucophyllum Extract against Oral Candidiasis: In Vitro and In Vivo Study.

Oral candidiasis (OC) is a fungal infection caused by an opportunistic fungi Candida albicans, which is found in the normal flora of healthy people. In this study, we examined the anti-candidal effect of green synthesized silver nanoparticles using leaf extract of Erodium glaucophyllum (EG-AgNPs) against C. albicans in vitro and in vivo. EG-AgNPs were synthesized for the first time using E. glaucophyllum extract and characterized by imaging (transmission electron microscopy (TEM), UV-VIS spectroscopy, zeta potential, X-ray diffraction (XRD), Energy dispersive x-ray analysis (EDX), and Fourier transform infrared spectroscopy (FTIR). A mouse model of OC was used for in vivo study. The agar well diffusion method showed the anti-candidal activity of EG-AgNPs against C. albicans with MIC 50 µg/mL. EG-AgNPs inhibited the dimorphic transition of C. albicans and suppressed the formation of biofilm by 56.36% and 52%, respectively. Additionally, EG-AgNPs significantly inhibited the production of phospholipases and proteinases by 30% and 45%, respectively. EG-AgNPs cause cytoplasm disintegration and deterioration of cell wall as imaged by SEM and TEM. Interestingly, EG-AgNPs did not display any cytotoxicity on the human gingival fibroblast-1 HGF-1 cell line at MIC concentrations. Topical treatment of the tongue of the OC mouse model with EG-AgNPs showed significant reduction in candidal tissue invasion, less inflammatory changes, and no tissue modification, in association with marked low scare and hyphal counts as compared to control group. In conclusion, our data demonstrated the potent inhibitory action of EG-AgNPs on the growth and morphogenesis of C. albicans in vitro and in vivo. Thus, EG-AgNPs represent a novel plausible therapeutic approach for treatment of OC.

Ca19Zn2(PO4)14 Nanoparticles: Synthesis, characterization and its effect on the colonization of Streptococcus mutans on tooth surface

The main cause of tooth decay is the biofilm formation of Streptococcus mutans. This study aimed to investigate the effect of the zinc oxide nanoparticles (ZnO NPs), hydroxyapatite nanoparticles (Ca10(PO4)6(OH)2) (HAP NPs), zinc oxide/hydroxyapatite nanocomposites (ZnO/HAP NCs), and Zn-substituted hydroxyapatite nanoparticles (Ca19Zn2(PO4)14 NPs) on the growth and biofilm formation, bacterial adherence, and the expression of ftf and gtf genes in Streptococcus mutans. The nanostructures were prepared via simple and fast co-precipitation route. Twelve isolates of Streptococcus mutans collected from children with dental caries referred to the dental clinic of Kashan University of Medical Sciences. All S. mutans isolates were susceptible to Ampicillin. The mean MIC for ZnO NPs, HAP NPs, Ca19Zn2(PO4)14, and ZnO/HAP NCs were 118, 260, 70.6, and 994 µg/mL, respectively. All prepared nanostructures significantly reduced biofilm formation at MIC and sub-MIC concentrations (p < 0.01). In biofilm and cell culture treated with nanoparticles, the expression of ftf and gtfC genes decreased. Results were shown that IC50 for the Ca19Zn2(PO4)14 was 8.5, and for non-toxic concentration, was 0.065 µg/mL. The attachment rate to the denture surface and HGF1 cell line treated with the Ca19Zn2(PO4)14 NPs has decreased. The results showed that the Ca19Zn2(PO4)14 NPs has a better effect than the ZnO/HAP NCs. It can therefore be used as a coating on dental surfaces. Investigation of Ca19Zn2(PO4)14 NPs form for covering dental teeth surface recommended in future study.

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

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

The effect of photodynamic therapy on head and neck squamous cell carcinoma cell lines using spirulina platensis with different laser energy densities

BackgroundConsidering the anti-cancer properties of spirulina platensis (S. platensis), we aimed to investigate the effectiveness of this algae as a novel natural photosensitizer for photodynamic therapy (PDT) against oral and hypopharyngeal cancer cells. The appropriate laser energy density to apply during PDT was also determined. Methods and MaterialsCAL-27, FaDu and HGF cell lines were exposed to S. platensis with concentrations of 0.3 g/l and 0.6 g/l and were irradiated with 635 nm diode laser using 2, 4, 12, and 24 J/cm2 energy densities with constant power. MTT assay was performed to investigate cell viability and cytotoxicity after 24 h. The results were analyzed using two-way ANOVA and post hoc Tukey tests (P-value<0.05). Resultssurvival rate in CAL-27 (P-Value<0.001) and FaDu (P-Value<0.001) cell lines were significantly different following irradiation with various laser energy densities. Different concentrations of S. platensis had no significant effect on the viability of CAL-27 cells (P-Value=0.158) and FaDu cells (P-Value=0.072) and showed no significant cytotoxicity against HGF cells, with or without laser. ConclusionS. platensis could be considered as a novel safe and effective natural photosensitizer for cancer PDT with no cytotoxic effect on normal cells. When combined with laser using appropriate energy densities, it has the ability to induce death in oral and hypopharyngeal cancer cell lines.

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

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

New Evidence for Artemisia absinthium L. Application in Gastrointestinal Ailments: Ethnopharmacology, Antimicrobial Capacity, Cytotoxicity, and Phenolic Profile.

Artemisia absinthium L. (Asteraceae) is traditionally used for gastrointestinal ailments and disorders linked to numerous risk factors including microbial infections. We aimed to provide contemporary evidence for its ethnopharmacological use and determine its antimicrobial capacity and mode of action, cytotoxicity, and phenolic constituents. Ethnopharmacological survey was conducted using semistructured interviews. Antimicrobial and antibiofilm capacities were determined by microdilution/crystal violet assay, respectively. Modes of action tested include estimation of exopolysaccharide production (congo red binding assay) and interference with membrane integrity (crystal violet uptake and nucleotide leakage assay). Cytotoxicity was determined using crystal violet assay. Polyphenolic profiling was done by advanced liquid chromatography/mass spectrometry (UHPLC-LTQ OrbiTrap MS). Artemisia absinthium in Serbia is traditionally used for gastrointestinal disorders, among others. Further study revealed high antifungal capacity of herb ethanolic extract towards range of Candida species (MIC 0.5–1 mg/mL) along with promising antibacterial activities (MIC 0.25–4 mg/mL). Interference with membrane integrity could be observed as a possible antimicrobial mechanism. Antibiofilm potential can be considered as high (towards C. krusei) to limited (towards P. aeruginosa) and moderate based on reduction in exopolysaccharide content. In concentrations up to 400 µg/mL, no cytotoxicity was observed towards HaCaT and HGF-1 cell lines. Polyphenolic analysis revealed twenty-one different constituents. A. absinthium usage as a gastrointestinal ailment remedy has been confirmed in vitro by its antimicrobial capacity towards microorganisms whose presence is linked to the diseases and associated complications and noncytotoxic nature of the natural product. The observed activities could be attributed to the present phenolic compounds.

Porphyromonas gingivalis lipopolysaccharide and gingival fibroblast augment MMP-9 expression of monocytic U937 cells through cyclophilin A.

Intercellular cross-talking was suggested in matrix metalloproteinase (MMP)-9 expression with unknown mechanisms. Studies showed cyclophilin A (CypA) playing an important role in regulating MMP-9 expression in varied diseases. The aim of the study was to examine the CyPA on the MMP-9 augmentation in monocytic U937 cells after Porphyromonas gingivalis (Pg) lipopolysaccharide (LPS) treatment and human gingival fibroblast (hGF) co-culture. In independent culture or co-culture of hGF and U937 cell, quantitative real-time polymerase chain reaction (qPCR) and zymography were selected to examine the mRNA and protein activity of MMP-9, respectively. The CyPA expression was determined by qPCR. LPS could enhance MMP-9 mRNA expression and enzyme activity in U937 cell. However, the enhancements were not observed in hGF. Similarly, LPS enhanced CyPA mRNA in U937, but not in hGF. After co-cultured with hGF, however, MMP-9 and CyPA in U937 increased regardless of the presence/absence of LPS. In U937 cells, the extra-supplied CyPA increased MMP-9 mRNA and enzyme activity, whereas the CyPA inhibitor, cyclosporine A, suppressed the LPS- and co-culture-enhanced MMP-9. Moreover, the inhibitors for MAP kinase, including PD98059 (ERK) and SP600125 (JNK), suppressed the CyPA-enhanced MMP-9 in U937. Through the CyPA pathway, the LPS and the hGF could augment the MMP-9 expression in the U937 cells.