Effect of alendronate on healing of bone defect and gingival tissue in osteopenic rats / Efecto del alendronato en la reparación de defectos óseos y tejido gingival en ratas con osteopenia

The aim was to evaluate bone repair and gingival tissue repair in osteopenic rats. Fifteen female wistar rats were included; in all of them ovariectomy was realized to induce osteopenia; after 45 days, the animals were submitted to 2 surgical techinques 1) dental extraction of the upper central incisor with no socket preservation and 2) 5 mm cranial defect in the calvarium; 5 rats were included in the control group (G1) withput alendronate application; in the group 2 (G2) was used subcutenous alendronate (0.5 mg/kg) once for three weeks and then was realizd the both surgical techniques. In group 3 (G3), after ovariectomy was realized the both dental extraction and the calvarium defect and after that was realized the alendronate protocol. In each group, after six week was realized euthanasia and descriptive histological analysis of the surgical areas involved. In bone formation of the 5 mm cranial defect was observed with good progression in the 3 experimental models and no modification in quality of bone repair was observed. For the gingival tissue in the extraction socket, no differences were observed between G1 and G3. On other hand, in G2 a thinner and reduced gingival epithelium was found. Our results showed that alendronate was not an obstacle for bone repair; deficiencies in re-epithelialization of oral mucosa show the impact of alendronate before dental extraction.

El objetivo fue evaluar la reparación ósea y gingival en ratas con osteopenia. Quince ratas wistar hembras fueron incluidas; en todas ellas se realizo ovarectomia y fue realizada la inducción de osteopenia; después de 45 días, los animales fueron sometidos a dos técnicas quirúrgicas 1) extracciones dentales del incisivo central superior sin preservación alveolar y 2) creación de un defecto craneano de 5 mm en la calota; 5 animales fueron incluidos como grupo control (G1) sin la aplicación de alendronato; en el grupo 2 (G2) se utilizó alendronato subcutáneo (0,5 mg/kg) una vez a la semana durante 3 semanas. En el grupo 3 (G3), después de la ovarectomia se realizó la exodoncia y el defecto en el cráneo y después de ello se inicio el protocolo con alendronato. En cada grupo, después de seis semanas se realizó la eutanasia con descripción histológica de los hallazgos. En el hueso formado en el defecto craneano de 5 mm se observó una adecuada progresión de reparación en los 3 modelos experimentales y no se observó cambios importantes en el modelo de reparación. Para el tejido gingival en el sitio de extracción, no se observaron diferencias entre el grupo G1 y G3. Por otra parte, el G2 presentó un tejido mas delgado con reducción del epitelio gingival; nuestros resultados demuestran que el alendronato no fue un obstáculo en la reparación ósea; deficiencias en la re epitelización de la mucosa oral muestran el impacto del alendronato después de la exodoncia.

Pam2CSK4 (TLR2 agonist) induces periodontal destruction in mice

Abstract Lipoproteins are important bacterial immunostimulating molecules capable of inducing receptor activator of nuclear factor-κB (RANKL) and osteoclast formation in vitro and in vivo . Although these molecules are present in periodontopathogenic bacteria, their role in periodontitis is not known. In this study, we used Pam2CSK4 (PAM2), a synthetic molecule that mimics bacterial lipoprotein, to investigate the effects of lipoproteins on periodontitis in mice. C57BL/6 male mice were randomly divided into three experimental groups: 1) Negative control group: animals received vehicle injection; 2) Positive control group: animals received injection of Escherichia coli lipopolysaccharide (LPS); 3) PAM2 group: animals received PAM2 injection. All the injections were performed bilaterally every other day into the palatal mucosa between first and second molars. After twenty-four days, the animals were euthanized to assess alveolar bone volume (micro-CT), cellular and extracellular composition in the gingiva (stereometric analysis), and osteoclast numbers (TRAP staining). Treatment with either PAM2 or LPS induced gingival inflammation, as demonstrated by increased infiltration of inflammatory cells and enhanced angiogenesis, associated with a smaller number of fibroblasts and decreased extracellular matrix. Importantly, treatment not only with LPS but also with PAM2 resulted in a larger number of TRAP+ multinucleated osteoclasts and significant loss of alveolar bone. Collectively, our data demonstrate that PAM2 can induce gingival inflammation and bone loss in mice, broadening the avenues of investigation into the role of lipoproteins in the pathogenesis of periodontal disease.

Pam2CSK4 (TLR2 agonist) induces periodontal destruction in mice

Abstract Lipoproteins are important bacterial immunostimulating molecules capable of inducing receptor activator of nuclear factor-κB (RANKL) and osteoclast formation in vitro and in vivo . Although these molecules are present in periodontopathogenic bacteria, their role in periodontitis is not known. In this study, we used Pam2CSK4 (PAM2), a synthetic molecule that mimics bacterial lipoprotein, to investigate the effects of lipoproteins on periodontitis in mice. C57BL/6 male mice were randomly divided into three experimental groups: 1) Negative control group: animals received vehicle injection; 2) Positive control group: animals received injection of Escherichia coli lipopolysaccharide (LPS); 3) PAM2 group: animals received PAM2 injection. All the injections were performed bilaterally every other day into the palatal mucosa between first and second molars. After twenty-four days, the animals were euthanized to assess alveolar bone volume (micro-CT), cellular and extracellular composition in the gingiva (stereometric analysis), and osteoclast numbers (TRAP staining). Treatment with either PAM2 or LPS induced gingival inflammation, as demonstrated by increased infiltration of inflammatory cells and enhanced angiogenesis, associated with a smaller number of fibroblasts and decreased extracellular matrix. Importantly, treatment not only with LPS but also with PAM2 resulted in a larger number of TRAP+ multinucleated osteoclasts and significant loss of alveolar bone. Collectively, our data demonstrate that PAM2 can induce gingival inflammation and bone loss in mice, broadening the avenues of investigation into the role of lipoproteins in the pathogenesis of periodontal disease.

Effects of 1, 25-dihydroxyvitamin D3 on experimental periodontitis and AhR/NF-κB/NLRP3 inflammasome pathway in a mouse model

Abstract Vitamin D has been known to have important regulatory functions in inflammation and immune response and shows inhibitory effects on experimental periodontitis in animal models. However, the potential mechanism has yet to be clarified. Recent studies have highlighted Aryl hydrocarbon receptor (AhR) and its downstream signaling as a crucial regulator of immune homeostasis and inflammatory regulation. Objective: This study aimed to clarify the effect of 1,25-dihydroxyvitamin D3 (VD3) on experimental periodontitis and AhR/nuclear factor-κB (NF-κB)/NLR pyrin domain-containing 3 (NLRP3) inflammasome pathway in the gingival epithelium in a murine model. Methodology: We induced periodontitis in male C57BL/6 wild-type mice by oral inoculation of Porphyromonas gingivalis (P. gingivalis), and subsequently gave intraperitoneal VD3 injection to the mice every other day for 8 weeks. Afterwards, we examined the alveolar bone using scanning electron microscopy (SEM) and detected the gingival epithelial protein using western blot analysis and immunohistochemical staining. Results: SEM images demonstrated that alveolar bone loss was reduced in the periodontitis mouse model after VD3 supplementation. Western blot analyses and immunohistochemical staining of the gingival epithelium showed that the expression of vitamin D receptor, AhR and its downstream cytochrome P450 1A1 were enhanced upon VD3 application. Additionally, VD3 decreased NF-κB p65 phosphorylation, and NLRP3, apoptosis-associated speck-like protein, caspase-1, interleukin-1β (IL-1β) and IL-6 protein expression. Conclusions: These results implicate the alleviation of periodontitis and the alteration of AhR/NF-κB/NLRP3 inflammasome pathway by VD3 in the mouse model. The attenuation of this periodontal disease may correlate with the regulation of AhR/NF-κB/NLRP3 inflammasome pathway by VD3.

Effects of cyclosporin, nifedipine and phenytoin on gingival myofibroblast transdifferentiation in monkeys

Abstract Objective: Myofibroblasts have been associated with the development of several pathologic fibrotic conditions. This longitudinal study aims to assess the proliferative and antiapoptotic effects of cyclosporin, nifedipine and phenytoin on gingival connective tissue cells of nonhuman primate, as well as to analyze a possible role of myofibroblasts in gingival overgrowth. Materials and Methods: Gingival samples from the right superior canine area were obtained from 12 male monkeys ( Sapajus spp ) to comprise the control group. After one week, the animals were randomly assigned to three groups, which received daily oral doses of cyclosporin, nifedipine or phenytoin for 120 days. Gingival samples were collected from the left superior canine area of two animals of each group at 52 and 120 days. Histological sections were stained with hematoxylin and eosin, and immunoreacted against α-SMA, Ki- 67 and bcl-2. Results: α-SMA immunoreaction was negative in the control and experimental groups. Similarly, no difference between groups concerning immunostaining against Ki-67 and bcl-2 was observed in connective tissue cells. Conclusion: Based on this methodology, it may be concluded that gingival overgrowths induced by cyclosporin, nifedipine and phenytoin are not associated with neither myofibroblast transdifferentiation, proliferation nor apoptosis of gingival connective cells in monkeys.

Effect of topical ozonetherapy on gingival wound healing in pigs: histological and immuno-histochemical analysis

Abstract In this study, the effects of ozonetherapy on secondary wound healing were evaluated histologically and immuno-histochemically. Material and Methods: 8 healthy pigs were used in this study. Six wounds with 10 mm in diameter were created through the punch technique on the palatinal gingiva of each pig. Ozone gas was applied on only 3 wounds (test group) and the remaining 3 were left to natural healing (control group). Biopsy samples were taken from one of the wounds in each group on the third day, from another wound of each group on the seventh day, and from another one on the tenth day. Routine histological analysis and immuno-histochemical staining were performed to investigate transforming growth factor-beta (TGF-β) and (VEGF) expressions. Results: No statistical difference was found between the test and control groups in terms of collagen fibers, epithelial formation and inflammation scores. A VEGF expression found in the test group was statistically higher than control group samples taken on the 3rd and 7th day. There was no statistical difference between the test and control groups in terms of TGF-β expression on any of the sampling days. Conclusion: The topical application of ozone gas could be effective in the early stages of wound healing by increasing the amount of VEGF expression. Clinical Relevance: Topical application of ozone gas may be effective in the early stages of oral wound healing.

Duration of effectiveness of Botulinum toxin type A in excessive gingival display: a systematic review and meta-analysis

Abstract Botulinum toxin type A is effective in reducing excessive gingival display caused by hyperfunctional upper lip elevator muscles; however, this effect is transient. This study aimed to determine the duration of the effectiveness of botulinum toxin type A on a gummy smile. A systematic search was conducted using Medline (PubMed), Scopus, and Web of Science electronic databases, from 1970 to March 2017 with no language restriction; the search included studies evaluating adult patients with excessive gingival display who were treated with botulinum toxin and were followed-up for at least 3 months. OpenGrey and Clinical Trial Registry were also consulted. Quality assessment was applied to determine the level of evidence and bias, and a meta-analysis was performed. Of 2181 full texts, 71 were obtained, with 3 prospective studies meeting the selection criteria. The gingival display was significantly reduced to baseline with 2, 4, and 8 weeks of treatment. The gingival display considerably reduced at the baseline-2-week comparison (-4.44 mm using raw data and-4.05 mm using the standard difference) and increased throughout the weeks of follow-up. There is scant evidence to determine the duration of the effectiveness of toxin type A on a gummy smile. The effect tends to be stable until at least 8 weeks of follow-up, and the gingival exposure may not return to baseline within 12 weeks of follow-up. Well-designed randomized clinical trials with a minimum of 6 months of follow-up are necessary to strengthen the evidence.

Cytidine-phosphate-guanosine oligodeoxynucleotides in combination with CD40 ligand decrease periodontal inflammation and alveolar bone loss in a TLR9-independent manner

Abstract Local administration of toll-like receptor 9 (TLR9), agonist cytidine-phosphate-guanosine oligodeoxynucleotide (CpG ODNs), and CD40 ligand (CD40L) can decrease ligature-induced periodontal inflammation and bone loss in wild type (WT) mouse. Objective: This study aimed to explore whether such effect is dependent on TLR9 signaling. Material and Methods: Purified spleen B cells isolated from WT C57BL/6J mice and TLR9 knockout (KO) mice were cultured for 48 hours under the following conditions: CD40L, CpG+CD40L, CpG at low, medium and high doses. We determined B cell numbers using a hemocytometer at 24 h and 48 h. Percentages of CD1dhiCD5+ B cells were detected by flow cytometry. Interleukin-10 (IL-10) mRNA expression and protein secretion were measured by quantitative real-time polymerase chain reaction (qRT-PCR) and by ELISA, respectively. The silk ligature was tied around the maxillary second molars for 14 days, during which the CpG+CD40L mixture or PBS was injected into palatal gingiva on days 3, 6, and 9. Results: For both WT and TLR9 KO mice, CpG significantly induced B cell proliferation, increased IL-10 mRNA expression and protein secretion of IL-10 but reduced CD1dhiCD5+ B cells population; local injection of CpG+CD40L mixture significantly decreased alveolar bone loss and the number of TRAP-positive cells adjacent to the alveolar bone surface, and significantly increased the gingival mRNA expression of IL-10 and decreased RANKL and IFN-γ mRNA expression. Conclusions: These results indicated that CpG plus CD40L decreased periodontal inflammation and alveolar bone loss in a TLR9-independent manner in ligature-induced experimental periodontitis.

Comparison of primary human gingival fibroblasts from an older and a young donor on the evaluation of cytotoxicity of denture adhesives

Abstract Denture adhesives (DA) improve the retention and stability of ill-fitting dentures, especially for older adults. These materials should be biocompatible, i.e., they cannot cause undesired biological responses and be non-cytotoxic to oral tissues. However, in vitro testing of DA biocompatibility employing primary cell culture may possibly be affected by other factors, such as the donor age. Objective To compare the cytotoxicity of three different denture adhesives when assessed in primary gingival fibroblasts from a young donor or from an older donor, as well as the release of the basic fibroblast growth factor (bFGF), and the inflammatory response marker interleukin-6 (IL-6). Material and Methods Gingival fibroblasts isolated from a 30- and a 62-year-old donor were assayed for proliferation (1-7 days) and sensitivity to latex (positive control). Fibroblasts were indirectly exposed to Corega Ultra (cream), Corega powder and Fixodent Original for a 24 h period and assayed by XTT and Crystal Violet tests. The release of IL-6 and bFGF by exposed cells was determined by ELISA. Results While cells from the young donor presented higher cell growth after 7 days, the sensitivity to increasing concentrations of latex extracts was very similar between young and older cells. Both XTT and CVDE detected no difference between the DA and the control group. All materials induced higher levels of IL-6 and bFGF compared to control. Cells from the older donor exposed to Corega Ultra released lower levels of cytokine and growth factor. Conclusions All materials were considered non-cytotoxic, but affected cytokine and growth factor release. The biological differences found between fibroblasts from both donors could be due to individual or age-related factors. The authors suggest the use of cells from older donors on studies of dental products aimed at older patients, to better simulate their physiological response.

Treatment of experimental periodontal disease by laser therapy in simvastatin-modified rats

Abstract Low intensity laser can be used as a promising alternative in the treatment of periodontal disease. Objective The aim of this study was to evaluate low-level laser therapy (LLLT) as an adjuvant treatment for scaling and root planing (SRP) for the treatment of induced periodontitis in simvastatin-modified rats. Material and Methods A total of 180 rats were evenly divided into two groups: Veh - receiving oral administration of polyethylene glycol (vehicle); S - receiving oral administration of Simvastatin. Periodontal disease was induced in both groups at the first mandibular molar. After seven days, the ligature was removed and the animals were divided into subgroups according to the following local treatments: NT - no treatment; SRP - scaling and root planing and irrigation with saline solution; and LLLT ¬- SRP and laser irradiation (660 nm; 0.03 W; 4 J). Ten animals in each subgroup/local treatment were euthanized at 7, 15 and 30 days. Samples of gingival tissue were processed to analyze the tissue oxidative damage and radiographic analysis. Levels of oxidative stress were analyzed by the expressions of Tripeptideglutathione (TG), Malondialdehyde (MDA) and Carbonylated Proteins (CP). Results The animals in S group had higher levels of TG and lower levels of MDA and CP compared with Veh group (p<0.05). Radiographically, in the intragroup analysis Veh and S, LLLT showed lower bone loss (BL) compared with NT and SRP, in all experimental periods (p<0.01). In addition, a lower BL was observed for the animals of Veh group treated with LLLT compared with treatment SRP in the S group, in all experimental periods. Conclusion Within the limits of this study, we can conclude that LLLT was effective as adjuvant treatment for SRP protecting against the occurrence of oxidative tissue damages as well as for reducing alveolar bone loss in experimentally induced periodontitis simvastatin-modified rats.

Local application of statins in the treatment of experimental periodontal disease in rats

Abstract Objective The objective of this study was to evaluate the local effects of statins as adjuvants for treatment by scaling and root planing (SRP) of periodontal disease induced in rats. Material and Methods Ninety rats were used in the present experiment. Periodontal disease was induced in all animals using a cotton thread placed in the left first mandibular molar. After 7 days of induction, the bandage was removed and the animals were divided into three groups: 1) NT group (n=30), no treatment; 2) SRP group (n=30): SRP and irrigation with control gel; 3) S group (n=30) - SRP and irrigation with Simvastatin. Ten animals from each group were euthanized at 7, 15 and 30 days after treatment. Gingival biopsy specimens were processed to analyze the expression of matrix metalloproteinase 8 (MMP-8). The mandibles were removed and submitted to radiographic and laboratory processing for histometric analysis. Results The S group showed a significantly lower expression of MMP-8 compared to NT and SRP groups in all experimental periods. In the radiographic and histometric analyses between the groups, S group showed a significantly lower bone loss (BL) compared to NT and SRP groups in all experimental periods. Conclusions Within the limits of this study, it can be concluded that locally applied statin was effective as an adjuvant treatment for SRP in rats with induced periodontal disease.

Chlorhexidine with or without alcohol against biofilm formation: efficacy, adverse events and taste preference

Abstract In recent years, different chlorhexidine formulations have been tested, including an alcohol-free alternative, but the effect of this solution on early biofilm formation is not clear. A crossover, randomized, double-blind clinical trial was conducted to evaluate the effect of two chlorhexidine solutions against supra- and subgingival biofilm formation (NCT#02656251). Thirty-five participants were randomized and asked to rinse twice daily with 15 ml of an alcohol-containing 0.12% chlorhexidine solution, an alcohol-free 0.12% chlorhexidine solution, or placebo. The study was conducted in three experimental periods of 4 days each, with a 10-day washout between the periods. All the experimental periods followed the same protocol, except that the solutions were switched. Biofilm distribution was evaluated every 24 hours by the Plaque-Free Zone Index, during 96 hours. Adverse events were self-reported and sensory evaluation was performed using a hedonic scale. Compared to the placebo, the chlorhexidine solutions resulted in a significantly higher number of surfaces free of plaque over 96 hours (p < 0.01), and were able to prevent subgingival biofilm formation (p < 0.01). The alcohol-free chlorhexidine solution was associated with a lower incidence of adverse events, compared with alcohol-containing chlorhexidine (p < 0.05); it also received better sensory evaluation and acceptance by trial participants, compared with the alcohol-containing chlorhexidine (p = 0.007), and had a similar inhibitory effect on the formation of supra- and subgingival biofilms.

Effect of lectin (ScLL) on fibroblasts stimulated with LPS - an in vitro study

Abstract: The lectin (ScLL) extracted from the Synadenium carinatum plant has been evaluated as an immunomodulator in diseases such as asthma, neosporosis and leishmaniasis. However, it has not yet been evaluated in the oral cavity. This study evaluated the effect of ScLL on viability, proliferation and release of IL-10 in human gingival fibroblasts (HGF) stimulated with lipopolysaccharide (LPS). HGF were stimulated with LPS 1 µg/ml and treated with ScLL in concentrations of 10, 5 and 2 µg/ml for 1 and 5 h, and evaluated by flow cytometry for viability, apoptosis (initial/advanced) and necrosis. The supernatant was collected to detect release of IL-10 by ELISA. The proliferation was assessed with the BrdU assay. Positive control consisted of cells maintained in Dulbecco's Modified Eagles Medium (DMEM), and the negative control, of those kept in tap water. Data were analyzed by ANOVA and Dunnett's test (α = 0.05). No significant difference was found for ScLL concentrations regarding viability or initial and advanced apoptosis (p=0.455). All the groups, including the positive control, had a significantly lower necrosis parameter than negative control at 5 h (p < 0.001). No difference was found for proliferation among the experimental groups (p = 0.832). ScLL at 5 and 2 µg/ml resulted in a lower release of IL-10 than positive and negative controls at 5 h (p = 0.047). The results indicated that ScLL concentrations tested were not cytotoxic, and had no effect on proliferation and release of IL-10 parameters. A thorough understanding of ScLL, regarding its immunomodulatory potential, may open the door to new perspectives for dentistry.

Novel biological activity of ameloblastin in enamel matrix derivative

Objective Enamel matrix derivative (EMD) is used clinically to promote periodontal tissue regeneration. However, the effects of EMD on gingival epithelial cells during regeneration of periodontal tissues are unclear. In this in vitro study, we purified ameloblastin from EMD and investigated its biological effects on epithelial cells. Material and Methods Bioactive fractions were purified from EMD by reversed-phase high-performance liquid chromatography using hydrophobic support with a C18 column. The mouse gingival epithelial cell line GE-1 and human oral squamous cell carcinoma line SCC-25 were treated with purified EMD fraction, and cell survival was assessed with a WST-1 assay. To identify the proteins in bioactive fractions of EMD, we used proteome analysis with two-dimensional gel electrophoresis followed by identification with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Results Purified fractions from EMD suppressed proliferation of GE-1 and SCC-25. LC-MS/MS revealed that ameloblastin in EMD is the component responsible for inhibiting epithelial cell proliferation. The inhibitory effect of ameloblastin on the proliferation of GE-1 and SCC-25 was confirmed using recombinant protein. Conclusion The inhibitory effects of EMD on epithelial cell proliferation are caused by the biological activities of ameloblastin, which suggests that ameloblastin is involved in regulating epithelial downgrowth in periodontal tissues. .

Avaliação de nova técnica não cirúrgica para tratamento de deficiência de papila em área estética: estudo clínico randomizado controlado / New evaluation technique non-surgical for papilla deficiency treatment in cosmetic area: randomized controlled trial

A ausência ou perda da papila interdental cria deficiência estética, problemas fonéticos, impactação alimentar e gera muita expectativa ao paciente. Até o momento, o tratamento da ausência ou perda da papila interdental tem sido mal sucedido e não há estudos que indiquem que a regeneração da papila é um resultado previsível. O objetivo deste estudo foi avaliar a efetividade da injeção de gel de ácido hialurônico de origem não animal na redução ou eliminação da deficiência de papila entre dentes naturais comparativamente ao tratamento por meio de enxerto de tecido conjuntivo subepitelial. Foram avaliados neste estudo 20 sítios de 6 pacientes de ambos os sexos, com idade variável de 29 a 62 anos, apresentando deficiência de papila entre dentes naturais, na região anterior superior, em pelo menos dois dentes. Os 20 sítios tratados foram aleatoriamente divididos em dois grupos, de acordo com o tratamento para correção da deficiência de papila por meio de enxerto de tecido conjuntivo subepitelial (grupo controle) ou por meio de injeção de gel de ácido hialurônico (grupo teste). Um examinador único, calibrado, avaliou a distância da ponta da papila ao ponto de contato com auxílio de sonda periodontal milimetrada antes e aos 1, 3 e 6 meses após o tratamento. Além disso, foram investigados, nos sítios tratados, as medidas de profundidade de sondagem, nível de inserção, índice de sangramento do sulco, índice de placa, distância do ponto de contato à crista óssea alveolar, distância da ponta da papila à crista óssea alveolar e largura da papila. Os resultados demonstraram que aos 6 meses de pósoperatório o percentual de mudança na altura da papila foi maior no grupo teste (14,94% ± 21,35%) do que no grupo controle (-1,39% ± 31,46%), entretanto sem diferenças significantes entre os grupos (p> 0.05). Não houve variação estatisticamente significante na largura da papila antes e aos 4 meses após o tratamento nos grupos teste (p= 0.09) e controle (p= 0.16), assim como não houve variação significativa na distância entre a ponta da papila e a crista óssea alveolar. Houve melhora significativa do Índice de Estética Rosa (IER) observado aos 6 meses de acompanhamento em comparação com a condição inicial no grupo teste (p= 0.0078; Wilcoxon), enquanto que não houve mudança significativa no IER observado no grupo controle aos 6 meses de acompanhamento (p= 0.35). Os resultados obtidos permitiram concluir que o tratamento da deficiência de papila por meio de injeção de gel de ácido hialurônico promove melhora da deficiência de papila, similar aos resultados obtidos com o tratamento por meio de enxerto de tecido conjuntivo subepitelial, porém com melhora estética significativa relacionada especialmente às características de cor e textura do tecido relativamente aos tecidos moles adjacentes.(AU)

The absence or loss of interdental papilla creates an esthetic deficiency, phonetic problems and food impaction and generates a lot of expectation for the patient. Until now, the treatment for absence or loss of interdental papilla is unsuccessful e and there are no researches that show that the papilla regeneration is a predictable outcome. The aim of this study was to evaluate the effectivity of a non-animal originated hyaluronic acid injection in the reduction or elimination of papilla deficiency between natural teeth in comparison to a sub epithelial connective tissue graft treatment. The analysis was made on 20 sites in 6 patients, both genders, 29 - 62 years, showing deficiency in the papilla between natural teeth in the upper anterior region in at least two teeth. The 20 sites treated were randomly divided into two groups, according to the treatment by subepithelial connective tissue graft (control group) or by hyaluronic acid injection (test group). A single calibrated examiner evaluated the distance between the tip of the papilla to the contact point using a graduated periodontal probe before the treatment and 1, 3 and 6 months after it. Besides, it were investigated probing pocket depth, clinical attachment level, gingival bleeding index, plaque index, distance from papilla to alveolar crest, distance from contact point to alveolar crest and width of the papilla. The results showed that 6 months after the procedure, the percentage of change in the papilla level was higher in the test group (14,94% ± 21,35%) than in the control group (-1,39% ± 31,46%), though not statistically significant (p>0.05). There was no significant difference variation in the width of the papilla before and 4 months after the treatment in test group (p=0.09) and control group (p=0.16), and there was no significant difference variation in the distance between the tip of the papilla and the alveolar bone crest. There was significant improvement of the Pink Esthetic Score (PES) after 6 months in comparison to the initial condition in test group (p=0.0078; Wilcoxon), while there was no significant difference in the PES in control group 6 months after treatment (p=0.35). The results allow to conclude that the treatment for of the papilla deficiency using hyaluronic acid injection promotes improvement, similar to the results of the sub epithelial connective tissue graft treatment, but with significant esthetic improvement related specially to the color and texture characteristics of the adjacent soft tissues.(AU)

Genotoxic potential of 10% and 16% Carbamide Peroxide in dental bleaching

Dental bleaching has become one of the most frequently requested esthetic treatments in dental offices. Despite the high clinical success observed with this procedure, some adverse effects have been reported, including a potential for developing premalignant lesions, root resorption and tooth sensitivity, especially when misused. The aim of this study was to evaluate the genotoxic response using a micronucleus (MN) assay, after the application of two concentrations of carbamide peroxide. Thirty-seven patients were divided into two groups and randomly received either a 10% carbamide peroxide (CP) (19) or a 16% carbamide peroxide (18) concentration for 21 days in individual dental trays. Gingival margin cells were collected immediately before the first use (baseline), and then 15 and 45 days after baseline. The cells were placed on a histological slide, stained by the Feulgen technique, and evaluated by an experienced blinded examiner. One thousand cells per slide were counted, and the MN rate was determined. The two groups were analyzed by the Wilcoxon rank-sum test and the Kruskal-Wallis equality-of-populations rank test. A slight increase in MN was observed for both groups, in comparison with the baseline, at 15 days. However, no difference was observed between the two groups (10% and 16%), at either 15 or 45 days (p = 0.90). When bleaching is not prolonged or not performed very frequently, bleaching agents containing carbamide peroxide alone will not cause mutagenic stress on gingival epithelial cells.

Efficacy of a triclosan formula in controlling early subgingival biofilm formation: a randomized trial

The aim of this study was to determine the efficacy of rinses with slurries of a dentifrice containing triclosan (TCS), as compared with rinses with slurries from a control dentifrice, in controlling early subgingival biofilm formation. A double-blind, randomized and cross-over clinical trial was designed, and 26 dental students were included. In the first period, participants were randomized to rinse with a TCS slurry or a control slurry, in a 12 h interval, and to refrain from mechanical cleaning. A Plaque Free Zone Index was assessed at 24 h, 48 h, 72 h and 96 h. After a washout period of 10 days, the second experimental period was conducted, following the same protocol as the first period, except that the slurry groups were switched. Use of the TCS slurry resulted in a significantly higher percentage of plaque-free surfaces, both at 24 h and at 72 h (p < 0.01). In the of 48-72 h interval, the triclosan slurry showed a lower percentage of sites converted to a score of 2 (38.1% for the testversus 40% for the control product, p = 0.015). In conclusion, rinsing with slurries of dentifrice containing TCS retards the down growth of bacterial biofilms from the supra- to the subgingival environment.

Clarifications, guidelines and questions about the dental bleaching "associated" with orthodontic treatment

With regard to the best moment for carrying out or recommending dental bleaching to orthodontic patients, some explanations and orientations are given in order to answers the following questions: 1) Why orthodontic treatment completion is considered the best opportunity for carrying out the procedure? 2) Why dental bleaching should not be performed immediately before orthodontic treatment? 3) If that would be possible at any special case, what would that be? 4) Why dental bleaching should not be performed during orthodontic treatment? 5) If that would be possible at any special case, what would that be? This article highlights why it is essential to protect both the mucosa and the cervical region, regardless of the moment when dental bleaching is performed, whether associated with orthodontic treatment or not. The "how", "why" and "if" of whether or not it is convenient to perform dental bleaching before orthodontic treatment are still a matter of clinical suggestion, as it is a procedure that is under analysis, empirical knowledge waiting for scientific proof or disproof! Although tooth enamel has adamantine fluid flowing within it, providing a specific metabolism that is peculiar to its own and which could scientifically explain and base the option of carrying out teeth whitening before and during orthodontic treatment, we must still be very careful.

Quanto ao melhor momento para se aplicar ou recomendar a clareação dentária aos pacientes ortodônticos, alguns esclarecimentos e orientações são explanados para responder questionamentos como: 1) Por que depois do tratamento ortodôntico se constitui a melhor oportunidade para tal procedimento?; 2) Por que não realizar a clareação dentária imediatamente antes do tratamento ortodôntico?; 3) Se poderia realizá-la em alguma condição especial, e qual seria?; 4) Por que não se deveria clarear os dentes durante o tratamento ortodôntico?; 5) Se possível em algumas situações especiais, quando seriam essas situações especiais? No presente artigo, se destacará porque é fundamental sempre proteger a mucosa e a região cervical, independentemente do momento em que se fizer uma clareação dentária relacionada ou não ao tratamento ortodôntico. O mecanismo de como, por que e se é ou não conveniente clarear os dentes antes da finalização dos tratamentos ortodônticos ainda representa uma sugestão clínica, um procedimento em análise e um conhecimento empírico à espera de sua comprovação ou desmitificação científica. Apesar do esmalte dentário ter uma circulação do líquido adamantino, que propicia um metabolismo próprio e específico, que pode vir a ser, cientificamente, a base para explicar e fundamentar a clareação dentária antes e durante o tratamento ortodôntico, ainda assim devemos ser muito cautelosos.

Effects of zinc during lactation on the junctional epithelium and inserted gum in rats / Efectos del zinc durante la lactancia sobre el epitelio de unión y la encía insertada en ratas

The purpose of this study was to evaluate the effects of zinc, during lactation, on the junctional epithelium and inserted gum of the first upper molar of rats. The study used one-day old male rats, divided into two groups: those whose mother had been treated with 300 mg zinc chloride (ZnCl2) in the drinker water (treated group), and those whose mothers did not receive ZnCl2 (control group). After 21 days, the rat pups were sacrificed. Using karyometrical techniques, the greater (D) and smaller (d) nuclear diameters of the different layers of the junctional and inserted gum epithelia were determined, and the mean geometric diameter, D/d ratio, perimeter, area, volume, volume/area ratio, eccentricity, shape coefficient, and the contour index were estimated. The 100-point Merz grid was used with the purpose of evaluating the citoplasmatic and celular volume, the nucleus/citoplasm relationship, number density, outer surface/basal layer ratio, the thickness of epithelial layers, and the surface density. The results were submitted to statistical analysis using the Wilcoxon-Mann-Whitney test. The nuclei of the studied structures were significantly smaller, and the stereological results demonstrated that there were smaller cells, hence meaning a greater number of cells per mm3 of tissue, in the treated group. Zinc caused changes on the studied epitheliums, according to morphometric and stereological evaluations.

El objetivo de este estudio fue evaluar los efectos del zinc durante la lactancia, sobre el epitelio de unión y la encía insertada del primer molar superior de ratas. Fueron utilizadas ratas macho de un día de edad, divididas en dos grupos: aquellas cuyas madres habían sido tratadas con 300 mg de cloruro de zinc (ZnCl2) con agua del bebedero (grupo tratado) y aquellas cuyas madres no recibieron ZnCl2 (grupo control). Las crías fueron sacrificadas después de 21 días. Utilizando técnicas cariométricas fueron medidos los diámetros mayor (D) y menor (d) de los núcleos de las células de los diferentes estratos del epitelio de unión y de la encía insertada, estimándose el diámetro geométrico medio, la relación D/d, perímetro, área, volumen, relación volumen/área, excentricidad, coeficiente de forma e índice de contorno. Fue usada la rejilla de Merz, de 100 puntos, con la finalidad de evaluar el volumen celular y citoplasmático, la relación núcleo/citoplasma, densidad numérica, relación superficie externa/superficie basal, espesor de las capas epiteliales y densidad de superficie. Los resultados fueron sometidos a análisis estadístico mediante el test de Wilcoxon-Mann-Whitney. En el grupo tratado los núcleos celulares de las estructuras estudiadas fueron significativamente menores y los resultados estereológicos demostraron que las células eran menores, por lo tanto, con mayor número por mm3 de tejido. De acuerdo a los resultados morfométricos y estereológicos, el zinc provocó cambios en los epitelios estudiados.

Efecto de los antagonistas de los receptores AT1 de la angiotensina II sobre la enfermedad periodontal inducida por inyecciones de lipopolisacáridos en ratas / Effect of ang II AT1 receptor antagonists, on periodontal disease induced by injections of lipopolysaccharide in rats

La Enfermedad Periodontal (EP) es una condición inflamatoria progresiva que afecta los tejidos que soportan y rodean a los dientes. Las endotoxinas bacterianas como los lipopolisacáridos (LPS), inducen una cascada inflamatoria causando resorción ósea mediante la producción y modulación de la red de citoquinas del tejido periodontal, del sistema RANK-RANKL-OPG y de la producción de especies reactivas del oxígeno (ERO). Siendo la vía final la activación del factor de transcripción NFκB para el control de la infección. Se sabe que el Sistema Renina Angiotensina (SRA) esta involucrado en la inflamación. Estas acciones pro-inflamatorias de la Ang II son producidas por la activación de NFkB mediante los receptores AT1, y por la generación de ERO. Nuestro objetivo fue determinar si la inhibición del receptor AT1 con el uso del valsartán reduciría la respuesta inmunitaria innata inflamatoria en un modelo de EP inducida por las inyecciones de LPS en la encía de las ratas. Nuestros resultados demuestran que el Valsartán disminuyó la leucocitosis sistémica, la movilidad dentaria, atenuó la pérdida de peso corporal de las ratas, disminuyó la formación de enzimas antioxidantes y NOS, redujo la producción y liberación de citocinas pro inflamatorias y aumentó las citocinas antinflamatorias, disminuyó la activación de p-p38, p-NFkB y la expresión de los receptores TLR4. El Valsartán también revirtió los efectos del LPS sobre la resorción ósea ya que disminuyó el número de osteoclastos, la expresión de los receptores RANK/RANKL/OPG y la relación RANKL/OPG y aumentó los depósitos de calcio y colágeno. Los mecanismos por los cuales el Valsartán reduce los efectos inflamatorios producidos por el LPS no están claras, pero la interferencia del ensamblaje de la NAD(P)H oxidasa con apocinina y el Tempol, indica que el Valsartán puede interferir con los pasos para el reconocimiento de LPS y su asociación con TLR4. Concluyéndose que las ERO participan en la señalización intracelular de la ANG II, vía el AT1R. Este estudio ayuda a dilucidar el papel del SRA en procesos inflamatorios. Además contribuye con sus resultados a ofrecer una alternativa terapéutica en el tratamiento de la EP.