Search Results (179)

Electrospun poly(ε-caprolactone) (PCL) membranes incorporating Triticum vulgare extract (TVE) were developed as biomimetic scaffolds for periodontal regeneration. Using a ternary solvent system, two experimental formulations (µF-P10 and µF-P10T1) were fabricated and compared against a commercial dermal matrix. SEM analysis revealed bimodal fiber distributions (0.77–1.74 µm) and a surface porosity of 29.86% for TVE-loaded membranes, significantly higher than that of the commercial control (25.26%). FT-IR confirmed that the PCL chemical integrity was preserved, while mechanical testing showed that extract incorporation reinforced the matrix, increasing the Young’s modulus from 2.90 × 10³ Pa to 3.54 × 10³ Pa. UHPLC–MS identified ferulic acid as the primary bioactive component (90%), with release kinetics following a first-order model (R² = 0.998) over 48 h. Biological assays with human gingival fibroblasts (HGF) confirmed non-cytotoxicity (>70% viability). While both membranes supported healing, the µF-P10 formulation showed superior performance, with 80.2% proliferation and 60.6% wound closure, approaching control levels. These findings demonstrate that PCL-TVE electrospun scaffolds effectively combine favorable morphology and controlled release, offering a promising alternative for subepithelial connective tissue regeneration. Full article

Silver diamine fluoride (SDF) is widely used in pediatric dentistry for caries arrest; however, concerns exist regarding its cytotoxicity. Green-synthesized nano-silver fluoride (NSF) is a potential alternative to SDF, offering antimicrobial efficacy with improved biocompatibility. This study aimed to evaluate the in vitro safety profile of green-synthesized NSF with 5% (w/v) fluoride using Camellia sinensis extract and to compare it with 38% SDF + potassium iodide (KI) formulation in human gingival fibroblasts (HGFs). Eluates of NSF and SDF+KI were tested at serial concentrations of 5%, 1%, 0.1%, 0.01% and 0.005%. Cell viability was assessed after 24, 48, and 72 h using the MTT assay. Additionally, the formation of reactive oxygen species (ROS) in HGFs was detected through fluorescence microscopy. Exposure to 5% SDF+KI resulted in almost complete loss of cell viability at all time points, whereas NSF demonstrated significantly higher viability under the same conditions. Lower concentrations of both materials maintained acceptable biocompatibility. ROS analysis revealed increased oxidative stress in response to 5% SDF+KI, while NSF induced significantly lower ROS levels. NSF exhibited superior biocompatibility compared to SDF+KI, supporting its potential as a safer silver-based material for caries management. Further in vitro and in vivo studies are needed to confirm its clinical safety profile. Full article

Glyphosate is the most widely used pesticide globally, raising concerns about its environmental persistence and biological impacts. Therefore, monitoring pesticide use is essential for assessing agricultural practices and the risks to human health associated with chemical use. This research examined glyphosate contamination in water (40 samples) and soil (28 samples) from northeastern Romania, an important agricultural region. Glyphosate concentrations in environmental water and soil samples were quantified using a spectrophotometric method based on ninhydrin derivatization, with good linearity over the concentration range 1–30 µg/mL (R² = 0.9981). Glyphosate was detected at concentrations above the LOQ in one water sample. Also, the study proposes a UHPLC-MS/MS method for the confirmation of glyphosate presence in the analyzed sample. Additionally, this study contributes to the characterization of the toxicity profiles of glyphosate and a commercial glyphosate-based formulation (Roundup^®) in primary human gingival fibroblast (hGF) cell lines. The commercial product Roundup, containing glyphosate, exhibited cytotoxicity similar to that of the active compound at low and intermediate doses; a significant cytotoxic effect was observed at a maximum concentration of 1 mM, with prolonged exposure. These findings demonstrate minimal cytotoxicity under the examined conditions and underscore the need for dose- and time-dependent assessments to evaluate the biological impact of herbicide formulations. Full article

The surface characteristics of zirconia may influence both soft tissue response and bacterial colonization. This study evaluated the surface roughness and water contact angle of zirconia fabricated by additive manufacturing (material jetting, NPJ) and subtractive manufacturing (milling), and investigated human gingival fibroblast (HGF-1) viability and Streptococcus mutans (S. mutans) (ATCC 25175) adherence on these surfaces, as well as the possible correlation between roughness and bacterial adhesion. Sixty-four zirconia specimens (1 × 1 × 0.1 cm) were fabricated (n = 32 per group), sintered, and standardized by abrasive polishing. Surface roughness and contact angle were measured. Cell viability was assessed using an MTT assay at 24, 48, and 72 h. Bacterial adhesion was quantified after 24 and 48 h of incubation. Data were analyzed using two-way ANOVA, independent t-tests, and Pearson correlation (α = 0.05). No significant differences in HGF-1 viability were observed at 24 and 48 h; however, at 72 h, subtractively manufactured zirconia demonstrated higher cell viability than additively manufactured specimens (p < 0.001). S. mutans adhesion was significantly greater on additively manufactured zirconia at 24 h (p = 0.002), with no significant difference at 48 h. Manufacturing technique influenced surface properties and early bacterial adhesion. Both materials exhibited acceptable biocompatibility within the tested conditions. Full article

Soft-tissue management is critical for guided bone regeneration (GBR), yet conventional titanium meshes lack the ability to regionally regulate fibroblast behavior where opposite biological responses are needed. Here, we fabricated two femtosecond-laser patterned stripe topographies on titanium using a unidirectional scanning strategy with parameter tuning, generating LSFL with a periodicity of 820 ± 30 nm and micro-grooves with a periodicity of 4.7 ± 0.1 μm. Surface morphology and physicochemical properties were characterized by SEM/AFM, XPS, microhardness testing, and wettability measurements. Human gingival fibroblasts (HGF-1) were used to assess adhesion, cytoskeletal organization, spreading area, and proliferation (CCK-8). The submicron LSFL promoted robust fibroblast adhesion, aligned cytoskeletal organization, larger spreading areas, and higher proliferation, whereas the micro-groove surface markedly restricted spreading and was associated with poorer cytoskeletal organization and lower proliferation. Alternating patterned regions further demonstrated geometry-driven spatial selectivity, with preferential cell occupation on LSFL stripes. These findings support a fabrication-ready surface-engineering strategy to synchronize rapid soft-tissue sealing while restricting unwanted fibroblast advancement at defined regions, offering a promising route toward more predictable GBR outcomes. Full article

Objectives: Combining mechanical plaque control, the physical removal of oral biofilm, with chemical plaque control, the use of agents to inhibit biofilm formation, is effective in preventing periodontal disease. Chlorogenic acid (CGA) found in coffee beans has medicinal effects, such as anti-inflammatory and antibacterial properties. Periodontal pathogens are difficult to reach in certain areas with traditional self-care tools, such as toothbrushes. Additionally, the viscous biofilm is difficult to remove using mechanical plaque control alone. Therefore, this study aimed to evaluate the efficacy of CGA in chemical plaque control. Methods: The mRNA and protein expression of inflammatory cytokines in lipopolysaccharide (LPS)-stimulated human gingival fibroblasts (HGFs) and human periodontal ligament fibroblasts (HPDLs) in the presence of CGA were analyzed using reverse transcription-qPCR and enzyme-linked immunosorbent assay. Additionally, the proliferation levels of oral bacteria in the presence of CGA were evaluated. Results: CGA suppressed mRNA and protein expression levels of the inflammatory cytokines, interleukin (IL)-1β and IL-8, in HGFs and HPDLs stimulated with Porphyromonas gingivalis LPS. Furthermore, CGA inhibited bacterial proliferation of Streptococcus mutans, Aggregatibacter actinomycetemcomitans, P. gingivalis, and Fusobacterium nucleatum. Conclusions: This study demonstrated that CGA exhibits anti-inflammatory effects on gingiva and periodontal ligaments, and antibacterial effects against oral bacteria. These results indicate the potential application of CGA in chemical plaque control and suggest its use in preventing periodontal disease progression. Full article

Background/Objectives: Thymol (THY) is widely used in oral care products for its antimicrobial and anti-inflammatory activity, but data on its cytocompatibility, potential differential effects on oropharyngeal-derived cells, and mucosal irritation under prolonged exposure remain limited. This study evaluated THY’s effects on healthy human gingival fibroblasts (HGF-1) and pharyngeal carcinoma (Detroit-562) cells after 24 h exposure, together with its irritation potential in ovo. Methods: Cells were treated with THY (100–300 µM) for 24 h. Cellular viability (MTT), morphology, mitochondrial alterations (MitoTracker™/Hoechst 33342), mitochondrial membrane potential (JC-1), and apoptosis/necrosis (AO/PI) were assessed. Clonogenic assays evaluated long-term proliferative capacity. Lastly, irritation score was examined using the HET-CAM assay at 300 µM. Results: THY produced a dose-dependent viability decrease in both lines, with HGF-1 viability remaining ≥75% and Detroit-562 reduced to ~68% at 300 µM. Morphology, mitochondrial staining, JC-1 ratios, and AO/PI imaging showed progressive apoptotic features, more evident in Detroit-562 cells. Clonogenic capacity increased slightly in HGF-1 at 100 µM and declined to ~75% at 300 µM, whereas Detroit-562 colonies decreased from ~68% to ~40% across the dose range. Additionally, THY (300 µM) showed no irritation in the HET-CAM assay. Conclusions: THY demonstrated acceptable cytocompatibility in gingival fibroblasts, stronger inhibitory effects on carcinoma cells at higher concentrations, and no acute irritation in ovo. These findings support THY’s safe use within defined concentration limits and justify further evaluation in advanced oral tissue models. Full article

Background and Objectives: Soft-tissue healing, particularly rapid epithelialization, is a critical determinant of successful periodontal regenerative therapy. Fibroblast growth factor-2 (FGF-2) and enamel matrix derivative (EMD) are regenerative biomaterials used clinically. However, their comparative effects on gingival epithelial and fibroblast cell behavior remain unclear. The objective of this study was to examine the effects of FGF-2 on the proliferation, migration, and wound closure dynamics of human gingival epithelial-like cells (Ca9-22) and human gingival fibroblasts (HGF-1) and to compare its effects with those of EMD. Materials and Methods: Ca9-22 and HGF-1 cells were stimulated with FGF-2 (10 µg/mL) or EMD (100 µg/mL) or left unstimulated (control). Wound closure was assessed via scratch assay, migratory capacity via Transwell assay, and proliferation via automated cell counting at pre-defined time points. Results: In Ca9-22 cells, both FGF-2 and EMD significantly accelerated wound closure in a time- and concentration-dependent manner and markedly enhanced cell migration and proliferation compared to controls. EMD consistently induced a stronger migratory response. In HGF-1 cells, FGF-2 significantly advanced wound closure by day 5, whereas EMD induced a non-significant favorable trend. Both treatments significantly increased cell proliferation and migration of HGF-1 cells, with EMD yielding the highest migratory cell count. Conclusions: FGF-2 promotes gingival soft-tissue healing by enhancing epithelial-like cell and fibroblast migration and proliferation, supporting rapid epithelialization. EMD produced comparable wound-healing effects, indicating that the activation of both epithelial and mesenchymal cells is a central mechanism shared by distinct regenerative agents. Full article

Porphyromonas gingivalis plays a key role in periodontal disease and has been associated with several serious systemic diseases. Its fimbriae are a major virulence factor. We recently demonstrated the formation of bundles of long FimA fimbriae in strain ATCC 33277. Transmission (TEM) and scanning electron microscopy (SEM) were used to examine a collection of P. gingivalis strains representing all seven known FimA types (I, Ib, IIa, IIb, III–V) and a P. gulae strain (type A). Additionally, two P. gingivalis strains (ATCC 49417 and OMI 1127) were investigated in dual-species approaches together with Fusobacterium nucleatum or Streptococcus oralis as well as in co-culture with human gingival fibroblasts (HGFs). To evaluate the role of fimbriae accessory proteins FimCDE, proteomic analysis of outer membrane vesicles (OMVs) was performed. Bundling was confirmed to occur regardless of FimA type but was impaired by strong capsule formation. Furthermore, tubular and chain-like outer membrane extensions (OMEs) were identified in most strains examined, including P. gulae. For the first time, fimbriae-associated OMVs (FAVs) were observed. REM images suggest that bundled fimbriae, OMEs and FAVs form connections with F. nucleatum and S. oralis. Proteome analysis of OMV content revealed the ratios of FimA to accessory proteins to be approximately 13:1 for FimC and FimD and approximately 7:1 for FimE. The results imply more accessory proteins per fimbriae or shorter FimA fimbriae in OMVs than in cells. Since FimCDE are known to be responsible for the adhesion properties and autoaggregation of FimA fimbriae, we propose that they could also mediate the stability of bundled fimbriae and the binding of OMVs. Full article

Background and Objectives: Oropharyngeal squamous cell carcinoma (OPSCC) is a common type of head and neck cancer with a progressive incidence in recent years. The limitations and the side effects associated with the current treatments require new therapeutic alternatives. Silibinin (SIL) is a phytocompound with multifaceted properties that has demonstrated antitumor effects in several types of cancer. The aim of this study was to assess the potential anticancer effects of SIL in Detroit 562 human pharyngeal cancer cells, an ideal model for HPV-negative OPSCC. Materials and Methods: Detroit 562 cells and HGF-1- human gingival fibroblasts were used as experimental models. For the mechanistic investigations, different methods, such as MTT assay, bright field microscopy, immunofluorescence staining, and specific assays and kits were applied to quantify intracellular ROS production, activation of caspases, and the colony formation assay. Results: Treatment with SIL (25–200 µM) for 48 h induced a selective cytotoxic effect in Detroit 562 cancer cells, being minimally toxic to healthy cells. The cytotoxic mechanism of action was characterized by a decreased cell viability, morphological alterations, elevation of intracellular ROS, decreased mitochondrial potential, mitochondrial and nuclear dysmorphologies, activation of caspases 9 and 3/7 and apoptosis occurrence, and decreased long-term colony formation. Conclusions: These findings show that SIL could represent a potential alternative therapy for HPV-negative OPSCC by triggering mitochondrial apoptosis and exerting a decline in the colonogenicity of Detroit 562 cancer cells. Full article

Background/Objectives: The main objective of this study is to report the green synthesis of copper oxide nanoparticles (CuONPs) using an aqueous extract from Pelargonium x hortorum (P. hortorum) leaves. It also aims to evaluate its biological activity as well as assess its cytotoxic effects on human gingival fibroblasts (HGFs). Methods: Copper oxide nanoparticles (CuONPs) were synthesized through chemical precipitation using an aqueous extract from P. hortorum leaves. These CuONPs were characterized with various techniques, including UV–Vis, Fourier transform infrared (FT-IR), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and transmission electron microscopy (TEM). Results: The UV–Vis spectrum showed a characteristic absorption peak for CuONPs. FT-IR spectroscopy identified alkoxide and aromatic groups associated with flavonoids and phenolic compounds from P. hortorum. The Cu–O bond was also observed in the same analysis. XRD confirmed that the CuONPs had a monoclinic CuO structure and XPS revealed copper was in the Cu (II) oxidation state bonded to oxygen, consistent with CuO. The nanoparticles were spherical with an average size of 40–53 nm as shown by TEM. The biological activities of CuONPs were tested against Streptococcus mutans (S. mutans) and Candida albicans (C. albicans). The minimum inhibitory concentration (MIC) was 20 µg/mL. Cytotoxicity tests on human gingival fibroblasts (HGFs) after 24 h showed a non-linear, dose-dependent cell viability profile, indicating CuONPs did not exhibit cytotoxicity within the tested range and could even promote cell proliferation at low and intermediate concentrations. Conclusions: This study successfully synthesized CuONPs via a green method, highlighting its potential as a biocompatible antimicrobial and antifungal agent. Full article

Background and Objectives: The use of proanthocyanidins (PACNs) alongside standard periodontal treatment procedures can improve periodontal and peri-implant tissue healing. The present study aimed to evaluate the effect of different concentrations of Pelargonium sidoides root extract (PSRE) on periodontal tissue proliferation in comparison with chlorhexidine digluconate (CHX). Materials and Methods: A cell culture study was performed using human gingival fibroblast (HGF-1) and human periodontal ligament fibroblast (HPDLF) lines. The HGF-1 cell line was exposed to CHX (the gold standard treatment in periodontal diseases) and PSRE at concentrations of up to 800 μg/mL, which were compared with negative controls. HGF-1 viability and proliferation were evaluated using fluorescence tests and the PrestoBlue assay, respectively. In addition, the cell proliferation induction ability of PSRE was evaluated by treating HGF-1 and HPDLF cells with PSRE at 25 and 50 μg/mL concentrations and measuring the TGFβ-1 levels using TGFβ-1 ELISA. Results: When comparing the effects of the 25 μg/mL PSRE treatment to the control, a statistically significant difference in HGF-1 cell growth was observed (0.297 ± 0.048 (mean ± SE) and 0.203 ± 0.01, respectively; p = 0.006). The strongest cytotoxic effect on HGF-1 cells was observed with CHX (0.007 ± 0.006, p < 0.001 vs. control). The HGF-1 and HPDLF cells showed statistically significant increases in TGFβ-1 levels when treated with PSRE at 25 and 50 μg/mL compared with the control (352.38 ± 31.32 (mean ± SE) and 330.99 ± 26.53 versus 161.07 ± 15.11 in HGF-1 cells; 397.53 ± 18.1 and 399.91 ± 27.61 versus 137.7 ± 16.54 in HPDLF cells, p < 0.001). Additionally, no negative effects were detected at low PSRE concentrations (less than 100 μg/mL). Conclusions: The results of this study suggested that PACNs may promote HGF-1 and HPDLF cell proliferation. In contrast, CHX showed cytotoxic effects. Full article

Zoledronic acid (ZA), a nitrogen-containing bisphosphonate, is widely used to treat osteoporosis and bone metastases. However, its clinical application is limited by adverse effects, notably bisphosphonate-related osteonecrosis of the jaw (BRONJ), which is associated with cytotoxicity in oral mucosal cells. Polydeoxyribonucleotide (PDRN), a salmon sperm-derived DNA polymer with regenerative and anti-inflammatory properties, has shown therapeutic potential in tissue repair; however, its ability to mitigate ZA-induced cytotoxicity remains poorly understood. Here, we investigated the molecular mechanisms of ZA-induced toxicity in HGF-1 cells, a human gingival fibroblast line, and evaluated the protective effects of PDRN. ZA treatment (50 µM, 48 h) significantly inhibited HGF-1 cell growth, accompanied by reduced phosphorylation of protein kinase B (PKB) and signal transducer and activator of transcription 3 (STAT-3), along with increased phosphorylation of TANK-binding kinase 1 (TBK1). TBK1 silencing restored cell growth under ZA exposure, whereas silencing PKB or STAT-3 further suppressed cell growth even without ZA. Co-treatment with PDRN (100 µg/mL) effectively prevented and reversed ZA-induced HGF-1 cytotoxicity. Mechanistically, PDRN inhibited ZA-induced TBK1 phosphorylation and partially restored PKB phosphorylation, though it did not reverse the reduction in p-STAT-3. Additionally, ZA significantly elevated intracellular reactive oxygen species (ROS) levels at 8 h, which were attenuated by PDRN. The antioxidant N-acetylcysteine (NAC) similarly reduced ZA-induced ROS and p-TBK1 levels and improved cell growth, although it had limited effects on p-PKB at 8 h. Importantly, delayed PDRN treatment following ZA exposure reversed ZA-induced cell growth inhibition and TBK1 activation in a dose- and time-dependent manner. In summary, these findings demonstrate that ZA suppresses HGF-1 cell growth through ROS production, TBK1 activation, and inhibition of PKB and STAT-3, whereas PDRN counteracts these effects primarily by suppressing TBK1 activation and oxidative stress. Full article

Zirconia is widely used for customized implant abutments owing to its esthetics, strength, and biocompatibility; however, the optimal surface modification for soft-tissue sealing and bone metabolic remains uncertain. This study evaluated how glass–ceramic spray deposition (GCSD), with or without handheld nonthermal plasma (HNP), alters zirconia surface physiochemistry and cellular responses. Field-emission scanning electron microscopy/energy-dispersive X-ray spectroscopy, surface roughness (Ra), wettability, and surface free energy (SFE) were measured. Human osteoblast-like cells (MG-63) and human gingival fibroblasts (HGF-1) were used to assess attachment and spreading, metabolic activity, cytotoxicity, and inflammatory response (tumor necrosis factor-α, TNF-α) (α = 0.05). GCSD produced an interlaced rod- and needle-like glass–ceramic layer, significantly increasing Ra and hydrophilicity. HNP further reduced surface contaminants, increased SFE, and enhanced wettability. The combination of GCSD and HNP yielded the greatest attachment and spreading for both cell types, without increases in cytotoxicity or TNF-α. GCSD with HNP creates a hydrophilic, micro-textured, chemically activated zirconia surface that maintains biocompatibility while promoting early attachment and bone metabolic activity, supporting its application for zirconia implant abutments. Full article

Surface treatments play a crucial role in modifying the surface properties and biological performance of dental ceramics. This study investigated the effects of surface conditions on the wettability, cytocompatibility, and bacterial resistance of 4 mol% Y₂O₃-stabilized tetragonal zirconia polycrystal (4Y–TZP) and two lithium disilicate (Li₂Si₂O₅) glass ceramics (Amber^® Mill (AM) and Amber^® Mill Abut-Crown (AC)). Human gingival fibroblast (HGF-1) responses and biofilm formation on the machined, polished, and glazed samples were evaluated. The polished 4Y–TZP sample exhibited the highest water contact angle (WCA; 71.3°), while that of the AC samples decreased as the sample was machined (58.4°), polished (46.8°), and glazed (14.0°). The wettability, cytocompatibility, and bacterial resistance of the dental ceramics were significantly influenced by material type and surface condition. Among the surface-treated samples, the glazed specimens exhibited the lowest WCA and bulk density; thus, wettability is an important factor for cell proliferation and bacterial resistance. Among all samples, HGF-1 cells adhered well to the glazed ceramics and significantly proliferated over time. Particularly, the 4Y–TZP and AC glazed samples exhibited the lowest biomass and strong resistance to biofilm formation and bacterial adhesion. Thus, the glaze dramatically affected HGF-1 cell growth and antibiofilm formation. Full article