Clinical and molecular analysis in Papillon-Lefèvre syndrome.

Papillon-Lefèvre syndrome (PLS; MIM#245000) is a rare recessive autosomal disorder characterized by palmar and plantar hyperkeratosis, and aggressively progressing periodontitis leading to premature loss of deciduous and permanent teeth. PLS is caused by loss-of-function mutations in the CTSC gene, which encodes cathepsin C. PLS clinical expressivity is highly variable and no consistent genotype-phenotype correlation has been demonstrated yet. Here we report the clinical and genetic features of five PLS patients presenting a severe periodontal breakdown in primary and permanent dentition, hyperkeratosis over palms and soles, and recurrent sinusitis and/or tonsillitis. Mutation analysis revealed two novel homozygous recessive mutations (c.947T>C and c.1010G>C) and one previous described homozygous recessive mutation (c.901G>A), with parents carrying them in heterozygous, in three families (four patients). The fourth family presented with the CTSC c.628C>T mutation in heterozygous, which was inherited maternally. Patient carrying the CTSC c.628C>T mutation featured classical PLS phenotype, but no PLS clinical characteristics were found in his carrier mother. All mutations were found to affect directly (c.901G>A, c.947T>C, and c.1010G>C) or indirectly (c.628C>T, which induces a premature termination) the heavy chain of the cathepsin C, the region responsible for activation of the lysosomal protease. Together, these findings indicate that both homozygous and heterozygous mutations in the cathepsin C heavy chain domain may lead to classical PLS phenotype, suggesting roles for epistasis or gene-environment interactions on determination of PLS phenotypes.

Smoking habit modulates peri-implant microbiome: A case-control study.

BACKGROUND AND OBJECTIVE: Smoking is a recognized risk factor for peri-implant disease and leads to microbiological changes in mucositis and peri-implantitis. However, there is no knowledge about the impact of smoking in healthy peri-implant tissue. The aim of the study was to evaluate the microbiome in a peri-implant environment in smokers with healthy peri-implant conditions. METHODS: Peri-implant biofilm was collected around single clinically healthy, screwed-retained, teeth-surrounded implants in 12 non-smoker (NSMK) and 12 smoker (SMK) non-periodontitis subjects (no bleeding and probing depth <4 mm). Bacterial DNA was isolated and 16S ribosomal RNA gene libraries were sequenced using pyrosequencing, targeting the V3-V4 region. Datasets were processed using the Quantitative Insights into Microbial Ecology, Greengenes and the Human Oral Microbiome Database databases. RESULTS: An evident difference in the SMK peri-implant microbiome was observed compared to the NSMK microbiome, with a large abundance of species, even with a healthy peri-implant. The SMK core-microbiome showed an abundance of Fusobacterium, Tannerella and Mogibacterium, while the NSMK core revealed an abundance of Actinomyces, Capnocytophaga and Streptococcus, genera that are usually related to periodontal health. The microbiome inter-relationship was shown to be more inter-generic in SMK then in NSMK, indicating different microbiome cohesion. CONCLUSION: Smoking negatively affected the peri-implant microbiome, leading to a disease-associated state, even in clinically healthy individuals.

Luffa operculata at a late period of gestation dysregulates melatonin and cytokines interfering with weight of dams and their male offspring.

ETHNOPHARMACOLOGICAL RELEVANCE: The tea made with the fruits of Luffa operculata (L.) Cogn. (Cucurbitaceae; EBN) is popularly used as abortive. AIM OF THE STUDY: The present work aimed at accessing how the exposition of female Wistar rats to 1.0 mg/kg of EBN (experimental group, EG), or distilled water (control group, CG), by gavage, at gestational days (GD) 17-21 interfered with the reproductive performance, and with dams' behavior after weaning. MATERIALS AND METHODS: At post-natal day 2 (PND2), the number of male and female pups was evaluated, as well as their weight. After weaning (PND21), dams were euthanized, and their liver and kidneys were removed for histological and biochemical analyses, while the blood was used in the evaluation of cytokines IL-1α, IL-1ß, IL-6 and TNF-α, corticosterone, adrenocorticotrophic hormone, melatonin, AST, ALT and creatinine levels. RESULTS AND DISCUSSION: Dams that were treated with EBN showed an anxiety-like behavior, weight loss at the end of gestation and weight gain at weaning, accompanied with a significant decrease in pro-inflammatory cytokines and in the melatonin level. No significant histological or biochemical alterations have occurred in the liver or kidneys. The number of female pups was significantly higher in the EG. The male pups showed weight gain at PND60. CONCLUSION: The presence of cucurbitacins is probably involved in the dysregulations that were found, due to their polycyclic steroid triterpene structure.

Novel rare frameshift variation in aggressive periodontitis: Exomic and familial-screening analysis.

BACKGROUND: Aggressive periodontitis (AgP), currently periodontitis grade C, presents early onset, rapid progression, and a poorly established genetic association. Thus, this study aimed to identify genetic variants associated with AgP via whole exome sequencing (WES) through a familial screening approach. METHODS: WES was performed in two nuclear families, including a proband and a parent affected by AgP and an unaffected parent and sibling. Common variants among affected individuals, excluding those common to healthy people, from each family, composed the data set associated with AgP. In silico analysis evaluated the impact of each variant on protein structure and protein-protein interactions. Moreover, identified deleterious variants were validated in a populational analysis (n = 96). RESULTS: The missense single nucleotide variations (SNVs) rs142548867 in EEFSEC (c.668C>T), rs574301770 in ZNF136 (c.466C>G), and rs72821893 in KRT25 (c.800G>A) and the frameshift indels rs37146475 in GPRC6A (c.2323-2324insT) and c.1366_1372insGGAGCAG in ELN were identified in AgP and have a predicted functional impact on proteins. In silico analysis indicated that the indel in GPRC6A generates a loss of the C-terminal tail of the Gprca protein. Furthermore, this SNV was significantly associated with AgP in a population-based investigation. CONCLUSION: Novel frameshift variation in GPRC6A (c.2323-2324insT) was identified as a potential genetic alteration associated with AgP occurrence.

Histometric evaluation of bone around titanium implants with different surface treatments in rats exposed to cigarette smoke inhalation.

OBJECTIVES: There is a lack of histological information about the influence of cigarette smoke on bone around surface-treated implants. The aim of the present study was to test the influence of titanium surface treatment on osseointegration in animals that were exposed to intermittent cigarette smoke inhalation. MATERIAL AND METHODS: Twenty-two male Wistar rats were used. One tibia, chosen at random, received a machined titanium implant (MI) while the other received an aluminum oxide-blasted surface implant (ABI). The animals were randomly assigned to one of the following groups: Group 1 - control (n=11) and Group 2 - intermittent cigarette smoke inhalation (n=11). Sixty days after surgery, the animals were sacrificed. The degree of bone-to-implant contact (BIC), bone filling (BF) within the limits of the threads of the implants and bone density (proportion of mineralized bone in a 500-mum-wide zone lateral to the implant - BD) were measured in the cortical (zone A) and cancellous bone (zone B) areas. RESULTS: Data analysis showed significant differences when comparing the groups and implant surfaces in both zones for BIC (two-way ANOVA -P<0.05). The two groups presented higher BIC mean values for ABI, when compared with MI (P<0.05). In group 2, cigarette smoke inhalation negatively affected BF in both zones (P<0.05). Group 2 presented a significantly decreased BD in both zones (P<0.05). No statistically significant differences were observed between surfaces in any of the groups for BD. CONCLUSION: Within the limits of the present study, it can be concluded that the aluminum oxide blast surface treatment may increase the degree of BIC but cannot overcome the detrimental effect of tobacco smoke on bone around titanium implants.

Systemic treatment with resveratrol reduces the progression of experimental periodontitis and arthritis in rats.

Rheumatoid arthritis and periodontitis are chronic inflammatory diseases which has been closely associated due to the nature of immune-inflammatory imbalance response. Resveratrol is a naturall product with biological proprieties that may promote immunomodulatory effects on host response. This study investigated resveratrol continuous administration effect on experimental periodontitis and arthritis progression in rats. Thirty-five rats were assigned to the following groups: 1-experimental arthritis + experimental periodontitis + placebo (RA+EP +PL) (n = 12); 2 -RA+EP+ ibuprofen (RA+PE+IB) (n = 11); 3-RA+EP+ resveratrol (RA+PE+RSV) (n = 11). After euthanasia, the specimens were processed for morphometric analysis of bone loss, and the gingival tissue surrounding the first molar was collected for quantification of inflammatory markers using a Luminex/MAGpix assay and anti-citrullinated protein antibody (ACCPA) levels were measured by ELISA assay. Serum level of rheumatoid factor (RF) was measured by ELISA assay. Paw edema was analyzed using a plethysmometer. Higher bone loss was observed in PL group, when compared to IB and RSV groups. RSV group presented higher IL-4 concentration than PL and IB groups. Resveratrol reduced RF serum levels and both IB and RSV decreased ACCPA gingival levels. Besides, paw swelling level was significantly lower in IB and RSV groups in the 21th day and only in RSV group in the 28th day. Histological analyzes showed smooth articular surface and higher width of the subchondral cortical in RSV group. Resveratrol showed modulatory effect and seems to reduce the inflammatory signs of arthritis and articular damage throughout the time.

Impact of a triclosan-containing toothpaste during the progression of experimental peri-implant mucositis: Clinical parameters and local pattern of osteo-immunoinflammatory mediators in peri-implant fluid.

BACKGROUND: This study evaluated the influence of a triclosan-containing toothpaste in the profile of osteo-immunoinflammatory mediators in peri-implant crevicular fluid (PICF) and in clinical parameters during progression of peri-implant mucositis. METHODS: Twenty-two clinically healthy patients with an implant-supported single-unit crown were enrolled in this double-blind, randomized, crossover study carried out in two phases of 21 days each. During an experimental 3-week period of undisturbed plaque accumulation in the implants, patients were randomly assigned to use three times/day: triclosan (n = 11), triclosan/copolymer/fluoride toothpaste; or placebo (n = 11), fluoride toothpaste. After a professional prophylaxis, a washout period of 30 days was established. Clinical parameters and 15 osteo-immunoinflammatory mediators in the PICF were evaluated at baseline and at 3, 7, 14, and 21 days. RESULTS: Both groups showed increase in plaque index at implant sites from the 3rd until the 21st day (P < 0.05). Only triclosan treatment was able to avoid an increase in bleeding on probing (BOP) throughout the follow-ups (P > 0.05), whereas a significant intensification in BOP was observed from the 14th day in the placebo-treated sites (P < 0.05). Lower interleukin (IL)-10 concentrations were detected in the placebo group at the 21st day when compared with triclosan-treated implant sites (P < 0.05). IL-10 levels were reduced and IL-1ß concentrations were increased at 21 days when compared with baseline only in placebo-treated sites (P < 0.05). Osteoprotegerin levels significantly increased from the 14th until the 21st day only in triclosan-treated sites (P < 0.05). CONCLUSION: Triclosan-containing toothpaste controls clinical inflammation and interferes positively in the profile of osteo-immunoinflammatory mediators during progression of experimental peri-implant mucositis.

Periodontitis increases rheumatic factor serum levels and citrullinated proteins in gingival tissues and alter cytokine balance in arthritic rats.

This study investigated some immunological features by experimental periodontitis (EP) and rheumatoid arthritis (RA) disease interact in destructive processes in arthritic rats. Rats were assigned to the following groups: EP +RA; RA; EP; and Negative Control. RA was induced by immunizations with type-II collagen and a local immunization with Complete Freund's adjuvant in the paw. Periodontitis was induced by ligating the right first molars. The serum level of rheumatoid factor (RF) and anti-citrullinated protein antibody (ACCPA) were measured before the induction of EP (T1) and at 28 days after (T2) by ELISA assay. ACCPA levels were also measured in the gingival tissue at T2. The specimens were processed for morphometric analysis of bone loss, and the gingival tissue surrounding the first molar was collected for the quantification of interleukin IL-1ß, IL-4, IL-6, IL-17 and TNF-α using a Luminex/MAGpix assay. Paw edema was analyzed using a plethysmometer. Periodontitis increased the RF and ACCPA levels in the serum and in the gingival tissue, respectively. Besides, the level of paw swelling was increased by EP and remained in progress until the end of the experiment, when EP was associated with RA. Greater values of IL-17 were observed only when RA was present, in spite of PE. It can be concluded that periodontitis increases rheumatic factor serum levels and citrullinated proteins level in gingival tissues and alter cytokine balance in arthritic rats; at the same time, arthritis increases periodontal destruction, confirming the bidirectional interaction between diseases.

Does Lip-Repositioning Surgery Improve Long-Term Smile Outcome and Dental Esthetics in Patients With Excessive Gingival Display? A Review of the Current Literature.

Focused Clinical Question: In patients with normal crown dimensions and excessive gingival display, does lip-repositioning surgery improve long-term smile outcome and dental esthetics? Clinical Scenario: A 27-year-old systemically healthy female presents with the chief complaint of "excessive gingival display when smiling" (Fig. 1). She reports that orthodontic treatment was performed and that she was referred for orthognathic surgery to correct the maxillary vertical excess and altered passive eruption. Clinically, the patient is periodontally healthy with favorable long-term periodontal prognosis. She is unsatisfied with her smile and states that it is "compromising her self-esteem." However, she refuses to accept the orthognathic surgery because of the extensiveness and morbidity of the procedure. She inquires about alternative solutions for her esthetic problem and whether it is possible to solve her gummy smile with a less invasive procedure. Lip-repositioning and anatomic crown-lengthening surgeries are presented as treatment options. The patient decides to pursue these treatment options. Lip-repositioning surgery is performed as described by Rosenblatt and Simon1 and Simon et al.,2 and healing is uneventful. Anatomic crown lengthening is performed 6 months after the previous surgery. The patient is pleased with the reduction of the excessive gingival display after these procedures (Fig. 2). However, there is lack of clinical evidence regarding the long-term stability of this surgical procedure.

Outcome of enamel matrix derivative treatment in the presence of chronic stress: histometric study in rats.

BACKGROUND: Psychologic stress and clinical hypercortisolism have been related to direct effects on bone metabolism. However, there is a lack of information regarding the outcomes of regenerative approaches under the influence of chronic stress (CS). Enamel matrix derivative (EMD) has been used in periodontal regenerative procedures, resulting in improvement of clinical parameters. Thus, the aim of this histomorphometric study is to evaluate the healing of periodontal defects after treatment with EMD under the influence of CS in the rat model. METHODS: Twenty Wistar rats were randomly assigned to two groups; G1: CS (restraint stress for 12 hours/day) (n = 10), and G2: not exposed to CS (n = 10). Fifteen days after initiation of CS, fenestration defects were created at the buccal aspect of the first mandibular molar of all animals from both groups. After the surgeries, the defects of each animal were randomly assigned to two subgroups: non-treated control and treated with EMD. The animals were euthanized 21 days later. RESULTS: G1 showed less bone density (BD) compared to G2. EMD provided an increased defect fill (DF) in G1 and higher BD and new cementum formation (NCF) in both groups. The number of tartrate-resistant acid phosphatase-positive osteoclasts was significantly higher in G1 when compared to G2 and in EMD-treated sites of both groups. CONCLUSIONS: CS may produce a significant detrimental effect on BD. EMD may provide greater DF compared to non-treated control in the presence of CS and increased BD and NCF in the presence or absence of CS.

Diabetes mellitus may increase bone loss after occlusal trauma and experimental periodontitis.

BACKGROUND: Diabetes mellitus (DM) involves metabolic changes that can negatively influence periodontal tissues, resulting in more prevalent and severe periodontitis and impaired bone formation. Occlusal trauma (OT) is an injury of the supportive periodontium that results in bone loss. It can be hypothesized that DM would increase bone loss after OT, mainly when associated with periodontitis. Thus, the aim of the present study is to evaluate the influence of DM on bone response in the furcation area of teeth subjected to OT in the presence, or absence, of experimental periodontitis (EP) in the rat model. METHODS: Thirty-two male Wistar rats were assigned to four groups: 1) group 1 (G1): DM+OT+EP (n = 8); 2) group 2 (G2): DM+OT (n = 8); 3) group 3 (G3): OT+EP (n = 8); and 4) group 4 (G4): OT (n = 8). G1 and G2 received a single intraperitoneal injection of streptozotocin (STZ). After 10 days, G1 and G3 were subjected to EP by ligature placement. Fifteen days after the start of EP, OT was induced by the creation of a premature contact. The animals were euthanized 35 days after DM induction. RESULTS: DM enhanced bone loss in the presence of OT combined with EP, but did not increase bone loss in teeth subjected to OT alone. EP caused greater bone loss when associated with OT. CONCLUSION: Within the limits of this animal study, it can be concluded that DM enhances bone loss in the presence of occlusal trauma associated with EP.

The impact of cigarette smoke inhalation on the outcome of enamel matrix derivative treatment in rats: histometric analysis.

BACKGROUND: Tobacco use is the most significant risk factor of periodontal disease. Clinical evidence has demonstrated that tobacco may negatively influence the results after surgical and non-surgical periodontal therapy. Enamel matrix derivative (EMD) have been used in periodontal regenerative procedures resulting in improvement of clinical parameters. The effect of EMD in the presence of tobacco compounds is unclear. Thus, the aim of the present study is to evaluate the impact of cigarette smoke inhalation (CSI) on the results of EMD treatment. METHODS: Twenty-two Wistar rats were assigned to two groups: Group 1, CSI (n = 11); Group 2, non-exposed (n = 11). Thirty days after initiation of CSI, fenestration defects were created at the buccal aspect of the first mandibular molar. The study followed a split-mouth design. After the surgeries the defects were randomly assigned to two subgroups: non-treated control and treated with EMD. The animals were sacrificed 21 days later and the percentage of defect fill, density of newly formed bone, and new cementum formation were histometrically assessed. The number of osteoclasts was determined by tartrate-resistant acid phosphatase. RESULTS: CSI was associated with less bone density compared to the non-exposed group. EMD provided an increased defect fill and new cementum formation in both groups. The number of tartrate-resistant acid phosphatase-positive osteoclasts was significantly higher in the CSI non-treated control group compared to the non-treated control of the non-exposed animals. CONCLUSIONS: EMD may provide increased defect fill and cementum formation in the presence or absence of CSI. However, tobacco smoke produced a detrimental effect on bone healing when density of newly formed bone was considered.

Estado atual da clorexidina na terapia periodontal / Current status of chlorhexidine in periodontal therapy

A clorexidina é uma bisbiguanida de amplo espectro que age em bactérias gram-positivas, gram-negativas, alguns vírus, leveduras e fungos. É considerada o agente químico gold standard no controle do biofilme supragengival e da inflamação e vem sendo utilizada como terapia coadjuvante em Periodontia desde a década de 1970. A literatura ao longo desse tempo vem propondo indicações, protocolos, formulações e concentrações distintas, podendo gerar dúvidas quanto a sua prescrição. Esta revisão teve como objetivo abordar a literatura científica com intuito de auxiliar o decision making do periodontista quanto ao uso da clorexidina frente ao tipo de doença, perfil sistêmico e limitações de cada paciente.