Effects of three restorative techniques in the bond strength and nanoleakage at gingival wall of Class II restorations subjected to simulated aging

Objective: To evaluate the effects of simulated aging in bond strength and nanoleakage of class II restorations using three different restorative techniques. Materials and methods: Class II preparations (n = 12) were restored using: FS - composite resin Filtek Supreme Plus (3M/ESPE); RMGIC + FS - resin-modified glass ionomer cement Vitrebond Plus (3M/ESPE) + FS; and FFS + FS - flowable composite resin Filtek Supreme Plus Flowable (3M ESPE) + FS. The teeth were assigned into two groups: Control and Simulated Aging - Thermal/Mechanical cycling (3,000 cycles, 20-80 °C/500,000 cycles, 50 N). From each tooth, two slabs were assessed to microtensile bond strength test (μTBS) (MPa), and two slabs were prepared for nanoleakage assessment, calculated as penetration along the restoration margin considering the penetration length (%) and as the area of silver nitrate particle deposition (μm2). Data were analyzed by two-way analysis of variance (ANOVA) followed by Tukey's post hoc test (p < 0. 05). Results: FS presented the highest μTBS to dentin (22. 39 ± 7. 55 MPa) after simulated aging, while the presence of flowable resin significantly decreased μTBS (14. 53 ± 11. 65 MPa) when compared to no aging condition. Both control and aging groups of RMGIC + FS presented the highest values of silver nitrate penetration (89. 90 ± 16. 31 % and 97. 14 ± 5. 76 %) and deposition area (33. 05 ± 12. 49 and 28. 08 ± 9. 76 μm2). Nanoleakage was not affected by simulated aging. Conclusions: FS presented higher bond strength and lower nanoleakage and was not affected by simulated aging. Use of flowable resin compromised the bond strength after simulated aging. Clinical relevance: The use of an intermediate layer did not improve the dentin bond strength neither reduced nanoleakage at the gingival margins of class II restorations under simulated aging conditions. © 2012 Springer-Verlag.

Toll-like receptor 2 knockdown modulates interleukin (IL)-6 and IL-8 but not stromal derived factor-1 (SDF-1/CXCL12) in human periodontal ligament and gingival fibroblasts

Background: Fibroblasts are now seen as active components of the immune response because these cells express Toll-like receptors (TLRs), recognize pathogen-associated molecular patterns, and mediate the production of cytokines and chemokines during inflammation. The innate host response to lipopolysaccharide (LPS) from Porphyromonas gingivalis is unusual inasmuch as different studies have reported that it can be an agonist for Toll-like receptor 2 (TLR2) and an antagonist or agonist for Toll-like receptor 4 (TLR4). This study investigates and compares whether signaling through TLR2 or TLR4 could affect the secretion of interleukin (IL)-6, IL-8, and stromal derived factor-1 (SDF-1/CXCL12) in both human gingival fibroblasts (HGF) and human periodontal ligament fibroblasts (HPDLF). Methods: After small interfering RNA-mediated silencing of TLR2 and TLR4, HGF and HPDLF from the same donors were stimulated with P. gingivalis LPS or with two synthetic ligands of TLR2, Pam2CSK4 and Pam3CSK4, for 6 hours. IL-6, IL-8, and CXCL12mRNA expression and protein secretion were evaluated by quantitative polymerase chain reaction and enzymelinked immunosorbent assay, respectively. Results: TLR2 mRNA expression was upregulated in HGF but not in HPDLF by all the stimuli applied. Knockdown of TLR2 decreased IL-6 and IL-8 in response to P. gingivalis LPS, or Pam2CSK4 and Pam3CSK4, in a similar manner in both fibroblasts subpopulations. Conversely, CXCL12 remained unchanged by TLR2 or TLR4 silencing. Conclusion: These results suggest that signaling through TLR2 by gingival and periodontal ligament fibroblasts can control the secretion of IL-6 and IL-8, which contribute to periodontal pathogenesis, but do not interfere with CXCL12 levels, an important chemokine in the repair process.

Biostimulatory effects of low-level laser therapy on epithelial cells and gingival fibroblasts treated with zoledronic acid

Low-level laser therapy (LLLT) has been considered as an adjuvant treatment for bisphosphonate-related osteonecrosis, presenting positive clinical outcomes. However, there are no data regarding the effect of LLLT on oral tissue cells exposed to bisphosphonates. This study aimed to evaluate the effects of LLLT on epithelial cells and gingival fibroblasts exposed to a nitrogen-containing bisphosphonate - zoledronic acid (ZA). Cells were seeded in wells of 24-well plates, incubated for 48 h and then exposed to ZA at 5 μM for an additional 48 h. LLLT was performed with a diode laser prototype - LaserTABLE (InGaAsP - 780 nm ± 3 nm, 25 mW), at selected energy doses of 0.5, 1.5, 3, 5, and 7 J cm-2 in three irradiation sessions, every 24 h. Cell metabolism, total protein production, gene expression of vascular endothelial growth factor (VEGF) and collagen type I (Col-I), and cell morphology were evaluated 24 h after the last irradiation. Data were statistically analyzed by Kruskal-Wallis and Mann-Whitney tests at 5% significance. Selected LLLT parameters increased the functions of epithelial cells and gingival fibroblasts treated with ZA. Gene expression of VEGF and Col-I was also increased. Specific parameters of LLLT biostimulated fibroblasts and epithelial cells treated with ZA. Analysis of these in vitro data may explain the positive in vivo effects of LLLT applied to osteonecrosis lesions. © 2013 Astro Ltd.

MyD88 or TRAM knockdown regulates interleukin (IL)-6, IL-8, and CXCL12 mRNA expression in human gingival and periodontal ligament fibroblasts

Background: In a previous report, it was shown that Toll-like receptor (TLR) 2 knockdown modulates interleukin (IL)-6 and IL-8 but not the chemokine CXCL12, an important mediator with inflammatory and proangiogenic effects, in human gingival fibroblasts (HGF) and human periodontal ligament fibroblasts (HPDLF). This study investigates whether knocking down two important TLR adaptor molecules, such as myeloid differentiation protein 88 (MyD88) and TRIF-related adaptor molecule (TRAM), could affect mRNA expression of IL-6, IL-8, and CXCL12 in HGF and HPDLF. Methods: After small interfering (si) RNA-mediated silencing of MyD88 or TRAM, HGF and HPDLF were stimulated with Porphyromonas gingivalis (Pg) lipopolysaccharide (LPS) or two synthetic ligands of TLR2 (Pam2CSK4 and Pam3CSK4) for 6 hours. IL-6, IL-8, and CXCL12 mRNAs were evaluated by quantitative polymerase chain reaction. Results: Knockdown of MyD88 or TRAM partially impaired the IL-8 mRNA upregulation in both fibroblast subpopulations. Similarly, IL-6 upregulation was partially prevented by siMyD88 or siTRAM in HGF stimulated with Pg LPS, as well as in both fibroblast subtypes challenged with Pam2CSK4. Conversely, constitutive CXCL12 mRNA levels were upregulated by MyD88 or TRAM knockdown in non-stimulated cells. Conclusions: These results suggest that TLR adaptor molecules knockdown, such as MyD88 or TRAM, can decrease IL-6 and IL-8 mRNA and increase CXCL12 mRNA expression in HGF and HPDLF. This can be an important step for better understanding the mechanisms that control the inflammatory cytokine and chemokine expression, which in turn contributes to periodontal pathogenesis.

Randomized controlled clinical trial evaluating connective tissue graft plus resin-modified glass ionomer restoration for the treatment of gingival recession associated with non-carious cervical lesion: 2-year follow-up

Background: The aim of this clinical study is to evaluate the 2-year term results of gingival recession (GR) associated with non-carious cervical lesions (NCCLs) treated by connective tissue graft (CTG) alone or in combination with a resin-modified glass ionomer restoration (CTG+R). Methods: Thirty-six patients with Miller Class I buccal GR associated with NCCLs completed the follow-up. The defects were randomly assigned to receive either CTG or CTG+R. Bleeding on probing (BOP), probing depth (PD), relative GR, clinical attachment level (CAL), and cervical lesion height coverage were measured at baseline, 6 months, 1 year, and 2 years after treatment. Results: Both groups showed statistically significant gains in CAL and soft-tissue coverage. The differences between groups were not statistically significant in BOP, PD, relative GR, or CAL after 2 years. Cervical lesion height coverage was 79.31% ± 18.51% for CTG and 71.95% ± 13.25% for CTG+R (P >0.05). Estimated root coverage was 91.56% ± 11.74% for CTG and 93.29% ± 7.97% for CTG+R (P ≥0.05). Conclusions: Within the limits of the present study, it can be concluded that both procedures provide comparable soft tissue coverage after 2 years of follow-up.

Plaque control improves the painful symptoms of oral lichen planus gingival lesions. A short-term study

BackgroundLichen planus is a mucocutaneous disease with manifestation in the oral mucosa, the gingiva being one of the most affected regions. In some cases, the lesion may be painful and lead to fragility of the tissues, so that precise diagnosis and adequate treatment are indispensible factors for improving the clinical condition. The aim of this study was to evaluate the effectiveness of plaque control in the improvement of clinical features and painful symptoms of oral lichen planus with gingival involvement.MethodsTwenty patients diagnosed with gingival lichen planus confirmed by histopathological examination were selected. The patients were evaluated by a trained examiner, with regard to the clinical features of the lesions [Index of Escudier etal. (Br J Dermatol, 157, 2007, 765)]; painful symptoms (Visual Analog Scale); and periodontally, as regards the visible plaque and gingival bleeding indices. Periodontal treatment consisted of supragingival scaling and oral hygiene instruction, with professional plaque removal afterward for a period of 4weeks. The entire sample was evaluated at the baseline and at the conclusion of treatment, and the results were analyzed by the Wilcoxon nonparametric test.ResultsThe data demonstrated that the majority of patients were women (90%), with a mean age of 55.9years. Periodontal treatment resulted in statistically significant reduction (P<0.05) in the periodontal indices, with consequent improvement in the clinical features and painful symptoms of the lesions.ConclusionsIt was demonstrated that plaque control was effective in improving the clinical features and painful symptoms of oral lichen planus with gingival involvement.

Effects of low-level laser therapy on the proliferation and apoptosis of gingival fibroblasts treated with zoledronic acid

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Synergistic Anti-Inflammatory Activity of the Antimicrobial Peptides Human Beta-Defensin-3 (hBD-3) and Cathelicidin (LL-37) in a Three-Dimensional Co-Culture Model of Gingival Epithelial Cells and Fibroblasts

Given the spread of antibiotic resistance in bacterial pathogens, antimicrobial peptides that can also modulate the immune response may be a novel approach for effectively controlling periodontal infections. In the present study, we used a three-dimensional (3D) co-culture model of gingival epithelial cells and fibroblasts stimulated with Aggregatibacter actinomycetemcomitans lipopolysaccharide (LPS) to investigate the anti-inflammatory properties of human beta-defensin-3 (hBD-3) and cathelicidin (LL-37) and to determine whether these antimicrobial peptides can act in synergy. The 3D co-culture model composed of gingival fibroblasts embedded in a collagen matrix overlaid with gingival epithelial cells had a synergistic effect with respect to the secretion of IL-6 and IL-8 in response to LPS stimulation compared to fibroblasts and epithelial cells alone. The 3D co-culture model was stimulated with non-cytotoxic concentrations of hBD-3 (10 and 20 mu M) and LL-37 (0.1 and 0.2 mu M) individually and in combination in the presence of A. actinomycetemcomitans LPS. A multiplex ELISA assay was used to quantify the secretion of 41 different cytokines. hBD-3 and LL-37 acted in synergy to reduce the secretion of GRO-alpha, G-CSF, IP-10, IL-6, and MCP-1, but only had an additive effect on reducing the secretion of IL-8 in response to A. actinomycetemcomitans LPS stimulation. The present study showed that hBD-3 acted in synergy with LL-37 to reduce the secretion of cytokines by an LPS-stimulated 3D model of gingival mucosa. This combination of antimicrobial peptides thus shows promising potential as an adjunctive therapy for treating inflammatory periodontitis.

Association between post-orthodontic treatment gingival margin alterations and symphysis dimensions

Orthodontic therapy is known to be associated with the development of gingival recession. Several factors may be involved in the causal chain of this outcome, including anatomical and behavioral aspects. Among the anatomical aspects, the dimensions of the mandibular symphysis could play a predisposing role. This study evaluated the relationship between the mandibular symphysis dimensions prior to orthodontic therapy and the development of gingival recessions on the lower incisors and cuspids. Records from 189 orthodontically treated adolescents were evaluated, including radiographs, casts and intra-oral photographs. Symphysis dimensions were assessed by cephalograms. Gingival margin alterations were determined in photographs and cast models. Association between gingival margin alterations and symphysis dimensions was tested by chi-square (α=0.05). Occurrence of gingival recession increased after orthodontic therapy. No association was observed on average of symphysis dimensions and the occurrence of gingival recessions. It may be concluded that pretreatment symphysis dimensions may not be used as predictors of gingival recession after orthodontic therapy.

Comparison between periodontal disease and a gingival carcinoma with emphasis on radiographic imaging

The majority of published papers deal mainly with prevalence, pathogenesis and treatment of squamous cell carcinoma of the gingiva (SCCG). On the other hand, little is discussed about the comparison between periodontal disease and gingival carcinoma with emphasis on radiographic imaging. In this case report we discuss the importance of the radiographic aspects in inflammatory periodontal disease and SCCG. This case report shows the importance of differentiating a localized severe periodontal disease and SCCG considering the radiographic aspects of the inflammatory bone loss and tumoral bone loss. The oral health care providers need to be familiar with the radiographic imaging of periodontal disease and SCCG.

Clinical assessment of nifedipine-induced gingival overgrowth in a group of brazilian patients

Although it has been established that nifedipine is associated with gingival overgrowth (GO), there is little information on the prevalence and severity of this condition in the Brazilian population. The aim of this study was to assess the occurrence of nifedipine-induced GO in Brazilian patients and the risk factors associated using a Clinical Index for Drug Induced Gingival Overgrowth (Clinical Index DIGO). The study was carried out on 35 patients under treatment with nifedipine (test group) and 35 patients without treatment (control group). Variables such as demographic (age, gender), pharmacological (dose, time of use), periodontal (plaque index, gingival index, probing depth, clinical insertion level, and bleeding on probing), and GO were assessed. Statistical analysis showed no association between GO and demographic or pharmacological variables. However, there was an association between GO and periodontal variables, except for plaque index. According to our study, the Clinical Index DIGO can be used as a parameter to evaluate GO. Therefore, we conclude that the presence of gingival inflammation was the main risk factor for the occurrence of nifedipine-induced GO.

Effect of LPS treatment on the viability and chemokine synthesis by epithelial cells and gingival fibroblasts

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Green tea polyphenol epigallocatechin-3-gallate and cranberry proanthocyanidins act in synergy with cathelicidin (LL-37) to reduce the LPS-induced inflammatory response in a three-dimensional co-culture model of gingival epithelial cells and fibroblasts

Objectives: The human antimicrobial peptide cathelicidin (LL-37) possesses anti-inflammatory properties that may contribute to attenuating the inflammatory process associated with chronic periodontitis. Plant polyphenols, including those from cranberry and green tea, have been reported to reduce inflammatory cytokine secretion by host cells. In the present study, we hypothesized that A-type cranberry proanthocyanidins (AC-PACs) and green tea epigallocatechin-3-gallate (EGCG) act in synergy with LL-37 to reduce the secretion of inflammatory mediators by oral mucosal cells. Methods: A three-dimensional (3D) co-culture model of gingival epithelial cells and fibroblasts treated with non-cytotoxic concentrations of AC-PACs (25 and 50 mg/ml), EGCG (1 and 5 mg/ml), and LL-37 (0.1 and 0.2 mM) individually and in combination (AC-PACs + LL-37 and EGCG + LL-37) were stimulated with Aggregatibacter actinomycetemcomitans lipopolysaccharide (LPS). Multiplex ELISA assays were used to quantify the secretion of 54 host factors, including chemokines, cytokines, growth factors, matrix metalloproteinases (MMPs), and tissue inhibitors of metalloproteinases (TIMPs). Results: LL-37, AC-PACs, and EGCG, individually or in combination, had no effect on the regulation of MMP and TIMP secretion but inhibited the secretion of several cytokines. ACPACs and LL-37 acted in synergy to reduce the secretion of CXC-chemokine ligand 1 (GRO-a), granulocyte colony-stimulating factor (G-CSF), and interleukin-6 (IL-6), and had an additive effect on reducing the secretion of interleukin-8 (IL-8), interferon-g inducible protein 10 (IP-10), and monocyte chemoattractant protein-1 (MCP-1) in response to LPS stimulation. EGCG and LL-37 acted in synergy to reduce the secretion of GRO-a, G-CSF, IL-6, IL-8, and IP-10, and had an additive effect on MCP-1 secretion. Conclusion: The combination of LL-37 and natural polyphenols from cranberry and green tea acted in synergy to reduce the secretion of several cytokines by an LPS-stimulated 3D coculture model of oral mucosal cells. Such combinations show promising results as potential adjunctive therapies for treating inflammatory periodontitis.

Cytotoxic Effects of Zoledronic Acid on Human Epithelial Cells and Gingival Fibroblasts

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)