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Characterization of Human Gingival Fibroblasts on Zirconia Surfaces Containing Niobium Oxide.

ABSTRACT: It was indicated that tetragonal zirconia polycrystal (TZP) containing yttria (Y₂O₃) and niobium oxide (Nb₂O₅) ((Y,Nb)-TZP) could be an adequate dental material to be used at esthetically important sites. The (Y,Nb)-TZP was also proved to possess its osteogenic potential comparable with those conventional dental implant material, titanium (Ti). The objective of the current study was to characterize cellular response of human gingival fibroblasts (HGFs) to smooth and rough surfaces of the (Y,Nb)-TZP disc, which were obtained by polishing and sandblasting, respectively. Various microscopic, biochemical, and molecular techniques were used to investigate the disc surfaces and cellular responses for the experimental (Y,Nb)-TZP and the comparing Ti groups. Sandblasted rough (Y,Nb)-TZP (Zir-R) discs had the highest surface roughness. HGFs cultured on polished (Y,Nb)-TZP (Zir) showed a rounded cell morphology and light spreading at 6 h after seeding and its proliferation rate significantly increased during seven days of culture compared to other surfaces. The mRNA expressions of type I collagen, integrin α2 and β1 were significantly stimulated for the Zir group at 24 h after seeding. The current findings, combined with the previous results, indicate that (Y,Nb)-TZP provides appropriate surface condition for osseointegration at the fixture level and for peri-implant mucosal sealing at the abutment level producing a suitable candidate for dental implantation with an expected favorable clinical outcome.

SUBMITTER: Cho YD

PROVIDER: S-EPMC5512668 | biostudies-other | 2015 Sep

REPOSITORIES: biostudies-other

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Enhanced Biological Behavior of In Vitro Human Gingival Fibroblasts on Cold Plasma-Treated Zirconia.

Project description:ObjectiveTo evaluate whether atmospheric-pressure dielectric-barrier-discharge plasma treatment of zirconia enhances its biocompatibility with human gingival fibroblasts.Materials and methodsThe zirconia disks were divided into four groups and treated using helium atmospheric-pressure dielectric-barrier-discharge plasmas for 30, 60 or 90 s or left untreated. The surface morphology, wettability and chemical elements were analyzed. Fibroblasts density, morphology, morphometry and attachment-related genes expression were measured at different time points from 3 to 72 h.ResultsAfter plasma treatment, the surface morphology and roughness remained the same, while the contact angle decreased from 78.31° to 43.71°, and the surface C/O ratio decreased from 3.17 to 0.89. The surficial areas and perimeters of HGFs were increased two-fold in the treated groups at 3 h. Fibroblasts density increased on treated disks at all time points, especially the ones treated for 60 s. Attachment-related genes in the groups treated for 30 and 60 s were significantly higher at 3 and 24 h.ConclusionThe helium atmospheric-pressure dielectric-barrier-discharge plasma treatment enhances the biological behavior of fibroblasts on zirconia by increasing the expression of attachment-related genes within 24 h and promoting the cell density during longer culture times. Wettability of zirconia, an important physicochemical property, has a vital influence on the cell behaviors.

| S-EPMC4603669 | biostudies-literature

Hydrofluoric acid treatment of titanium surfaces enhances the proliferation of human gingival fibroblasts.

Project description:The attachment of implants relies on bone and soft tissue biocompatibility. The aim of this article is to investigate the effect of fluoride-modified metallic titanium (Ti) surfaces (Ti-F) on proliferation and differentiation of human gingival fibroblasts. Human gingival fibroblast cells were exposed to hydrofluoric acid-modified Ti coins (Ti-F) for 1, 3, 7, 14 and 21 days, and untreated coins were used as controls. A five- to six-fold increase in the proliferation of human gingival fibroblasts on Ti-F compared to Ti surfaces was observed. Enhanced gene expression of interleukin-6 and osteoprotegerin was found at 7 days. Increased levels of sclerostin, interleukin-6 and osteoprotegerin in the media from human gingival fibroblasts cultured on Ti-F coins were found compared to controls. Our results confirm that hydrofluoric acid-modified surface may indirectly enhance the firm attachment of implant surface to junction epithelium, soft tissue epithelium, which would give protection for underlying osseous structures making osseointegration of the dental implant possible.

| S-EPMC6378639 | biostudies-literature

CEMP1 Induces Transformation in Human Gingival Fibroblasts.

Project description:Cementum Protein 1 (CEMP1) is a key regulator of cementogenesis. CEMP1 promotes cell attachment, differentiation, deposition rate, composition, and morphology of hydroxyapatite crystals formed by human cementoblastic cells. Its expression is restricted to cementoblasts and progenitor cell subpopulations present in the periodontal ligament. CEMP1 transfection into non-osteogenic cells such as adult human gingival fibroblasts results in differentiation of these cells into a "mineralizing" cell phenotype. Other studies have shown evidence that CEMP1 could have a therapeutic potential for the treatment of bone defects and regeneration of other mineralized tissues. To better understand CEMP1's biological effects in vitro we investigated the consequences of its expression in human gingival fibroblasts (HGF) growing in non-mineralizing media by comparing gene expression profiles. We identified several mRNAs whose expression is modified by CEMP1 induction in HGF cells. Enrichment analysis showed that several of these newly expressed genes are involved in oncogenesis. Our results suggest that CEMP1 causes the transformation of HGF and NIH3T3 cells. CEMP1 is overexpressed in cancer cell lines. We also determined that the region spanning the CEMP1 locus is commonly amplified in a variety of cancers, and finally we found significant overexpression of CEMP1 in leukemia, cervix, breast, prostate and lung cancer. Our findings suggest that CEMP1 exerts modulation of a number of cellular genes, cellular development, cellular growth, cell death, and cell cycle, and molecules associated with cancer.

| S-EPMC4444236 | biostudies-literature

Nicotine inhibits myofibroblast differentiation in human gingival fibroblasts.

Project description:Cigarette smoking has been suggested as a risk factor for several periodontal diseases. It has also been found that smokers respond less favorably than non-smokers to periodontal therapy. Previous work in our lab has shown that nicotine inhibits human gingival cell migration. Since myofibroblasts play an important role in wound closure, we asked if nicotine affects gingival wound healing process by regulating myofibroblast differentiation. Human gingival fibroblasts (HGFs) from two patients were cultured in 10% fetal bovine serum cell culture medium. Cells were pretreated with different doses of nicotine (0, 0.01, 0.1, and 1 mM) for 2 h, and then incubated with transforming growth factor beta (TGF-beta1) (0, 0.25, 0.5, and 1 ng/ml) with or without nicotine for 30 h. The expression level of alpha-smooth muscle actin (alpha-SMA), a specific marker for myofibroblasts, was analyzed by Western blots, immunocytochemistry, and real-time polymerase chain reaction (real-time PCR). Phosphorylated p38 mitogen-activated protein kinase (Phospho-p38 MAPK) activity was analyzed by Western blots. TGF-beta1 induced an increase of alpha-SMA protein and mRNA expression, while nicotine (1 mM) inhibited the TGF-beta1-induced expression of alpha-SMA but not beta-actin. Nicotine treatment down-regulated TGF-beta1-induced p38 MAPK phosphorylation. Our results demonstrated for the first time that nicotine inhibits myofibroblast differentiation in human gingival fibroblasts in vitro; supporting the hypothesis that delayed wound healing in smokers may be due to decreased wound contraction by myofibroblasts.

| S-EPMC2862373 | biostudies-literature

Synthesis, Characterization, and Photonic Efficiency of Novel Photocatalytic Niobium Oxide Materials.

Project description:The application of niobium oxides as photocatalytic materials for the removal of contaminants is scarcely reported in the literature. This work reports the methodology to synthesize four different mesoporous niobium oxide materials and the correlation between the physicochemical properties and the photocatalytic activity. X-ray diffraction, UV-vis diffuse reflectance spectra (DRS), transmission electron microscopy, and nitrogen adsorption techniques are used to characterize the structure and composition of the obtained materials. The photocatalytic oxidation of methanol is used as reaction test to assess the photocatalytic activities and photonic efficiencies of the materials as a function of the catalyst concentration. Nb2O5 materials display lower reaction rates, which can be attributed to the relatively high average particle size. By contrast, NaNbO3 materials show higher activity, especially for high catalyst loading. No significant differences in absorption and scattering of light are observed among the materials, indicating that the higher photonic efficiency of NaNbO3 should be the result of a lower charge recombination derived from its microstructure, sodium composition, low particle size, and high specific surface area of these materials.

| S-EPMC6607167 | biostudies-literature

Expression of deubiquitinases in human gingiva and cultured human gingival fibroblasts.

Project description:BackgroundAlthough deubiquitinating enzymes (DUBs) such as CYLD, A20 and OTULIN are expressed in multiple tissues and thought to be linked with inflammatory diseases, their expression in periodontal tissues remains to be determined. This research was designed to assess the expression of CYLD, A20 and OTULIN in human gingiva, and to evaluate the regulation of these DUBs in human gingival fibroblasts (HGFs) upon different stimuli.MethodsImmunohistochemistry assay was conducted to determine the expression of CYLD, A20 and OTULIN in human gingiva. Immunofluorescence assay was employed to observe the protein expression of CYLD, A20 and OTULIN in HGFs. RT-PCR and western blots were carried out to assess gene and protein expression changes of these DUBs in HGFs upon LPS or TNF-α.ResultsCYLD, A20 and OTULIN were found to be expressed in human gingiva and HGFs. The expression of CYLD, A20 and OTULIN was lower in the inflamed gingival tissue samples compared with the healthy gingival tissue samples. Further, the expression of CYLD, A20 and OTULIN in HGFs exhibited distinct regulation by different stimuli. TNF-α treatment markedly increased NF-κB activation in HGFs CONCLUSIONS: Our findings suggest that CYLD, A20 and OTULIN might play a role in the progression of periodontitis.

| S-EPMC8180082 | biostudies-literature

Dicationic Imidazolium-Based Ionic Liquid Coatings on Zirconia Surfaces: Physico-Chemical and Biological Characterization.

Project description:In the present work, dicationic imidazolium-based ionic liquids (ILs) were investigated as multi-functional coatings on a zirconia (ZrO?) surface to prevent biofilm formation and enhance the wear performance of zirconia while maintaining the material's compatibility with host cells. ILs containing phenylalanine and methionine were synthesized and deposited on zirconia. Intermolecular interactions driving IL deposition on zirconia were studied using X-ray photoelectron spectroscopy (XPS). Anti-biofilm activity and cell compatibility were evaluated in vitro after one and seven days, and wear performance was tested using a pin-on-disk apparatus. ILs were observed to form strong hydrogen bonds with zirconia. IL containing phenylalanine formed a stable film on the surface after one and seven days in phosphate-buffered saline (PBS) and artificial saliva and showed excellent anti-biofilm properties against Streptococcussalivarius and Streptococcussanguinis. Compatibility with gingival fibroblasts and pre-osteoblasts was maintained, and conditions for growth and differentiation were preserved. A significantly lower coefficient of friction and wear volume loss were observed for IL-coated surfaces as compared to non-coated substrates. Overall, zirconia is an emerging alternative to titanium in dental implants systems, and this study provides additional evidence of the materials' behavior and IL coatings as a potential surface treatment technology for improvement of its properties.

| S-EPMC5748557 | biostudies-literature

Expression profile of human gingival fibroblasts induced by interleukin-1beta reveals central role of nuclear factor-kappa B in stabilizing human gingival fibroblasts during inflammation.

Project description:BackgroundInterleukin (IL)-1beta is a key cytokine in the pathogenesis of periodontitis, and it induces inflammatory mediators in periodontal diseases. We developed immortalized human gingival fibroblasts (HGFs), investigated the effects of IL-1beta on the gene expression using expression arrays containing approximately 40,000 genes, and tested the role of nuclear factor-kappa B (NF-kappaB) in maintaining an activated HGF population.MethodsTotal RNA was isolated from IL-1beta-induced and mock-induced control cells. Gene expression analyses were performed using expression arrays and confirmed by quantitative real-time polymerase chain reaction. Western blot analysis to show inhibitor of kappa B-alpha (IkappaBalpha) phosphorylation and immunostaining of cells for NF-kappaB nuclear translocation were performed. Apoptosis was confirmed by assay of poly ADP-ribose polymerase (PARP) cleavage.ResultsA total of 382 probe sets corresponding to 254 genes were differentially expressed in IL-1beta-induced cells (P <0.001). A total of 215 genes were upregulated, and 39 genes were downregulated. Most notable NF-kappaB pathway members (NFkappaB1, NFkappaB2, IkappaBalpha, IkappaBepsilon, IkappaBzeta, REL, RELB, and TA-NFKBH) were upregulated. IkappaBalpha was phosphorylated, and NF-kappaB accumulated in the nucleus. An IL-1beta-induced set of 27 genes was downregulated by an NF-kappaB inhibitor, leading to a decreased number of viable cells and suggesting an antiapoptotic role for NF-kappaB.ConclusionsIL-1beta leads to a large number of significant expression changes consistent with a pathologic role in periodontitis, including enhancement of inflammatory cytokines, chemokines, transcription factors, matrix metalloproteinases, adhesion molecules, and especially NF-kappaB-dependent antiapoptotic genes. NF-kappaB activation blocks apoptosis, thereby stabilizing the HGF population in inflammation.

| S-EPMC4150685 | biostudies-literature

Drug-Induced Gingival Overgrowth: The Effect of Cyclosporin A and Mycophenolate Mophetil on Human Gingival Fibroblasts.

Project description:Drug-induced gingival overgrowth may occur after a chronic administration of three classes of systemic drugs: Anticonvulsants, immunosuppressants, and calcium channel blockers. This study aimed to investigate how cyclosporin A and mycophenolate mophetil (immunosuppressive drugs) could interfere with human gingival fibroblasts functions, leading to gingival enlargement. Human gingival fibroblasts derived from the tissue of a 60-year-old female were cultured in a DMEME medium. A stock solution with 1 mg/mL of mycophenolate and 1 mg/mL of cyclosporine were prepared and dissolved in a DMEM medium to prepare a serial dilution at the concentrations of 5000, 2000, 1000, 500, and 100 ng/mL, for both treatments. Cell viability was measured using the PrestoBlue™ Reagent Protocol. Quantitative real-time RT-PCR was performed in order to analyze the expression of 57 genes coding for gingival fibroblasts "Extracellular Matrix and Adhesion Molecules". Mycophenolate and cyclosporine had no effect on fibroblast cell viability at 1000 ng/mL. Both the treatments showed similar effects on the expression profiling of treated cells: Downregulation of most extracellular matrix metalloproteases genes (MMP8, MMP11, MMP15, MMP16, MMP24) was assessed, while CDH1, ITGA2, ITGA7, LAMB3, MMP12, and MMP13 were recorded to be upregulated in fibroblasts treated with immunosuppressive drugs. It has been demonstrated that gingival overgrowth can be caused by the chronic administration of cyclosporin A and mycophenolate mophetil. However, given the contrasting data of literature, further investigations are needed, making clear the possible effects of immunosuppressive drugs on fibroblasts.

| S-EPMC7400382 | biostudies-literature

Biofunctionalization of zirconia with cell-adhesion peptides <i>via</i> polydopamine crosslinking for soft tissue engineering: effects on the biological behaviors of human gingival fibroblasts and oral bacteria.

Project description:Rapid soft tissue integration is essential for long-term dental implant success. Zirconia is increasingly used as an abutment material owing to its excellent aesthetic properties and biocompatibility; however, it is bioinert, and tissue integration is poor. We developed a feasible surface modification method, exploiting the reactivity of polydopamine (PDA) films to immobilize cell-adhesion peptides (Arg-Gly-Asp, RGD) onto zirconia abutment surfaces. Further, we evaluated the effect thereof on human gingival fibroblast (HGF) behavior and oral bacterial adhesion, which influence the peri-implant soft tissue seal. HGF responses to linear KGGRGDSP and cyclic RGDfK sequences were compared. PDA deposition and covalent coupling of RGD were verified by X-ray photoelectron spectroscopy and fluorescence microscopy. The biological behaviors of HGFs on the modified zirconia; i.e., adhesion, spreading, proliferation, gene and protein expression, were elucidated. Biofunctionalization of zirconia with the adhesion peptides significantly enhanced the biological activities of HGFs. Cyclic RGD induced slightly improved cell attachment, spreading, and proliferation, but similar cell differentiation when compared to linear RGD peptides. To assess their antimicrobial properties, the different substrates were exposed to cultures of the early colonizer Streptococcus mutans or the periodontal pathogen Porphyromonas gingivalis, and bacterial adhesion was evaluated by scanning electron microscopy and live/dead staining. PDA and PDA-RGD coatings decreased zirconia surface colonization by both bacterial species to similar extents. Thus, PDA-RGD-functionalized zirconia modulates specific HGF responses, while maintaining the antimicrobial activity of the PDA coating. The selective bio-interaction pattern of this surface modification holds great promise for improving soft-tissue integration around zirconia abutments in clinical applications.

| S-EPMC9049673 | biostudies-literature

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