Platelet-Rich Fibrin in Coverage of Gingival Recession: A Systematic Review and Meta-Analysis.

This systematic review and meta-analysis sought to evaluate the efficacy of platelet-rich fibrin (PRF) membranes versus subepithelial connective tissue grafts (SCTGs) in the coverage of Miller class I and II gingival recessions. After the inclusion and exclusion criteria were applied, the quality of seven articles (Cohen's Kappa = 0.9) was evaluated using the Jadad scale. The MEDLINE/PubMed, Cochrane, and Web of Science databases were consulted, and manual searches were performed in the most popular periodontics journals. The studies included considered a total of 122 patients, 203 surgical fields on which SCTGs were used, and 205 surgical fields on which PRF was used. The parameters analyzed were probing depth, clinical attachment level, gingival recession, and keratinized mucosa. The minimum follow-up period accepted was 6 months. A statistically significant difference between the SCTG and PRF groups was found only in the case of keratinized mucosa. However, gingival recession, clinical attachment level, and probing depth parameters in the PRF group were found to be statistically equal to those of the SCTG group (the gold standard) (p ≥0.05). PRF membranes were determined to be a promising alternative to autogenous gingival grafts in the treatment of Miller class I and II gingival recessions.

A new technique for tongue brushing and halitosis reduction: the X technique.

The tongue is one of the primary sources of halitosis. The manual or mechanical removal of biofilm is known to decrease oral malodor. OBJECTIVE: To evaluate a new tongue hygiene technique hereby referred to as "the X technique" and its effects on both halitosis and the number of microorganisms based on microbiological parameters and diagnostic features of the breath. MATERIAL AND METHODS: The study included thirty patients divided into a control group (patients without systematized guidelines of lingual hygiene, but who performed the mechanical cleaning of tongue dorsum, each in its own way), the 3R group (instructed to perform the movements of the X technique for 3 repetitions at each brushing), and the 6R group (instructed to perform 6 repetitions of the technique at each brushing). After two weeks, a new data collection was performed. RESULTS: Patients in the 6R group presented the lowest score on the organoleptic assessment scale at the second consultation, followed by the 3R group and the controls. Regarding the self-perception of breath by the method of Visual Analog Scale (VAS), the control group did not perceive improvements in oral malodor; the results of the 3R group and the 6R group were similar. CONCLUSION: These results indicate that the X technique improves both measurements and perceptions of halitosis. Microbiological analyses revealed greatest reduction in the 6R group. The findings show that the X technique reduces both organoleptic scores and the number of bacterial colonies, and improves users' perceptions of their breath.

Application of botulinum toxin type A in gummy smile: case report / Aplicação da toxina botulínica tipo A em sorriso gengival: relato de caso

ABSTRACT Gummy smile is one of the major complaints of patients, due to its influence on the self-esteem. It is known that it has a multifactorial cause and can be present in the excessive vertical growth of the maxilla, excessive labial contraction, short upper lip and extrusion of the anterior teeth. The use of the botulinum toxin can be associated with additional treatments or be applied individually, according to the need of each patient. The goal of this research study was to present a clinical case using botulinum toxin type A as an alternative therapy to correct the gummy smile. A 22-year-old patient, presenting vertical maxillary growth, reporting aesthetic discomfort while smiling, chose for an alternative treatment therapy with the application of botulinum toxin to correct the accentuated gingival display. The results show that the toxin is a satisfactory treatment option and its indication is considered a fast, effective, easy and safe treatment method, being a more conservative approach compared to surgical procedures.

RESUMO O sorriso gengival, é uma das grandes queixas dos pacientes, devido a sua influência na auto estima do mesmo. Sua causa é multifatorial, podendo estar presente no excesso de crescimento vertical da maxila, contração labial excessiva, lábio superior curto e extrusão dos dentes anteriores. A aplicação da toxina botulínica pode ser associada a outros tratamentos ou isoladamente, de acordo com a necessidade de cada paciente. O objetivo desse trabalho foi apresentar um caso clinico utilizando a toxina botulínica tipo A como uma alternativa terapêutica para a correção do sorriso gengival. Paciente, 22 anos, apresentando crescimento vertical da maxila, relatando desconforto estético ao sorrir, optou pela alternativa terapêutica, com a aplicação da toxina botulínica para correção da exposição gengival acentuada. Os resultados encontrados mostram que a toxina é um tratamento satisfatório e sua indicação é considerada um tratamento rápido, eficaz, fácil e seguro. Um método mais conservador quando comparado aos procedimentos cirúrgicos.

A new technique for tongue brushing and halitosis reduction: the X technique

Abstract The tongue is one of the primary sources of halitosis. The manual or mechanical removal of biofilm is known to decrease oral malodor. Objective: To evaluate a new tongue hygiene technique hereby referred to as "the X technique" and its effects on both halitosis and the number of microorganisms based on microbiological parameters and diagnostic features of the breath. Material and Methods: The study included thirty patients divided into a control group (patients without systematized guidelines of lingual hygiene, but who performed the mechanical cleaning of tongue dorsum, each in its own way), the 3R group (instructed to perform the movements of the X technique for 3 repetitions at each brushing), and the 6R group (instructed to perform 6 repetitions of the technique at each brushing). After two weeks, a new data collection was performed. Results: Patients in the 6R group presented the lowest score on the organoleptic assessment scale at the second consultation, followed by the 3R group and the controls. Regarding the self-perception of breath by the method of Visual Analog Scale (VAS), the control group did not perceive improvements in oral malodor; the results of the 3R group and the 6R group were similar. Conclusion: These results indicate that the X technique improves both measurements and perceptions of halitosis. Microbiological analyses revealed greatest reduction in the 6R group. The findings show that the X technique reduces both organoleptic scores and the number of bacterial colonies, and improves users' perceptions of their breath.

Biomateriais utilizados na artroplastia parcial ou total da articulação temporomandibular: uma revisão de literatura / Biomaterials used in partial or total temporomandibular joint arthroplasty: a review of the literature

Introdução: as disfunções da articulação temporomandibular constituem uma variedade de doenças que compreendem disfunções nos músculos da mastigação, nas estruturas articulares ou em ambos. Em alguns casos a melhora da disfunção pode decorrer apenas de tratamento clínico conservador, porém, quando os resultados não são favoráveis, o tratamento cirúrgico deve ser considerado. A reconstrução articular parcial ou total é uma das formas de tratamento cirúrgico disponível. Objetivo: objetivo deste trabalho foi realizar uma revisão bibliográfica sobre os principais biomateriais disponíveis para a reconstrução da articulação temporomandibular. Método: o trabalho foi realizado na base de dados da Pubmed, Scielo e Periódicos Capes. Os descritores empregados foram: artoplastia temporomandibular, prótese articular, disfunção temporomandibular. As referências foram avaliadas quanto à relevância correlacionadas ao tema proposto. Foram selecionados os artigos em português e inglês, publicados no período de Janeiro 1971 a outubro de 2017. Considerações finais: existe a recomendação de haver uma continuidade de esforços para melhorar o material aloplástico para que os resultados obtidos com essa técnica se tornem o tratamento padrão para a DTM severa, contudo é importante não se esquecer de esgotar as modalidades terapêuticas conservadoras antes de se prosseguir para qualquer tipo de cirurgia. Nos casos onde uma prótese articular total é indicada, que os princípios biomecânicos sejam respeitados e que estudos clínicos metodologicamente adequados sejam realizados para fundamentarem esta condição de maneira referendada para a substituição aloplástica da ATM.

Introduction: temporomandibular joint dysfunctions are a variety of diseases that include dysfunctions in masticatory muscles, joint structures, or both. In some cases the improvement of the dysfunction may only result from conservative clinical treatment, however, when the results are not favorable the surgical treatment should be considered. Partial or total joint reconstruction is one of the forms of surgical treatment available. Objective: the objective of this work was to perform a bibliographic review on the main biomaterials available for the reconstruction of the temporomandibular joint. Method: work was carried out in the database of Pubmed, Scielo and Capes Periodicals. The descriptors employed were: temporomandibular arthroplasty, joint prosthesis, temporomandibular dysfunction. The references were evaluated regarding the relevance correlated to the proposed theme. The articles in Portuguese and English, published between January 1971 and October 2017, were selected. Final remarks: there is a recommendation for continuity of efforts to improve the alloplastic material so that the results obtained with this technique become the treatment standard for severe TMD, but it is important to remember to exhaust the conservative therapeutic modalities before proceeding for any type of surgery. In cases where a total joint prosthesis is indicated, biomechanical principles are respected and methodologically adequate clinical studies are performed to support this condition in a manner recommended for the alloplastic replacement of TMJ.

Biomateriais utilizados em artroplastia de quadril / Biomaterials used in hip arthroplasty

Introdução: a artroplastia de quadril é um procedimento cirúrgico utilizado com frequência na recuperação do quadril lesado, sendo necessário a utilização de biomateriais na confecção das próteses que farão a nova função dessa articulação. Os biomateriais mais frequentemente utilizados em reconstruções de quadril podem ser agrupados em: metálicos, poliméricos, cerâmicos e compósitos. Objetivo: discorrer sobre tipos de biomateriais utilizados nos implantes das artroplastias de quadril, apresentando suas características, vantagens e falhas, descritas na literatura. Método: foi realizado uma revisão da literatura nas bases de dados eletrônicos: PubMED/Medline, Lilacs e Scielo. Conclusão: os materiais utilizados em artroplastia de quadril evoluíram muito nos últimos 20 anos, especialmente considerando a resistência ao desgaste. Porém, não é somente o tipo de material implantado que determina os resultados em artroplastia do quadril. Bons resultados dependem de uma cirurgia bem indicada, bem planejada, bem realizada e do uso de material adequado.

Introduction: hip arthroplasty is a surgical procedure frequently used in the recovery of the injured hip, and it is necessary to use biomaterials in the preparation of prostheses that will perform a new function of this joint. The biomaterials most used in hip reconstructions can be grouped into metallic, polymeric, ceramic and composite. Objective: to describe the types of biomaterials used in hip arthroplasty implants, presenting their characteristics, advantages and failures, descriptions in the literature. Method: a literature review was performed in the electronic databases: PubMED / Medline, Lilacs and Scielo. Conclusion: the materials used in hip arthroplasty have evolved a lot in the last 20 years, considering the resistance to wear. However, it is not the type of implanted material that determines the results in hip arthroplasty. Good results depend on well-indicated, well-planned, well-performed surgery and use of the right material.

Diagnóstico e tratamento cirúrgico do freio labial com inserção marginal: relato de caso / Diagnosis and surgical treatment of labial frenulum with marginal insertion: a case report

O freio labial é uma estrutura presente em todos os indivíduos que em alguns casos pode apresentar-se mal posicionado, gerando efeitos adversos importantes, como formação de diastemas, mal adaptação de prótese, inibição dos movimentos dos lábios e língua, inflamação gengival, retração gengival e perda óssea. O presente estudo foi realizado com objetivo de relatar o diagnóstico e tratamento de um caso clínico de freio labial inferior com inserção marginal, causando retração gengival. Paciente de 39 anos de idade, gênero feminino, apresentou-se com acúmulo de biofilme e retração gengival entre os incisivos centrais inferiores. Após o exame clínico e a constatação da presença de freio labial inferior com inserção na gengiva marginal e ausência de gengiva queratinizada, optou-se pelo tratamento cirúrgico por meio da frenectomia labial inferior. Após o período de 45 dias do procedimento cirúrgico, o processo de reparo estava adequado, a paciente não relatou sintomatologia dolorosa, não havia sinais de infecção e houve uma melhora na qualidade de gengiva inserida. Pode-se concluir que, desde que bem indicada, através de um diagnóstico correto, a frenectomia labial pode resultar em um bom prognóstico e melhora da qualidade da gengiva inserida. (AU)

The labial frenulumis a structure present in all individuals that in some cases may be poorly positioned, generating important adverse effects such as diastema formation, maladaptation of prosthesis, inhibition of lip and tongue movements, gingival inflammation, gingival retraction and bone loss. The present study was carried out with the objective of reporting the diagnosis and treatment of a clinical case of lower labial brake with abnormal insertion, causing gingival retraction. A 39-year-old female patient presented with biofilm accumulation and gingival retraction between the lower central incisors. After the clinical examination and observation of the presence of a lower labial frenulum with marginal gingival insertion and absence of keratinized gingiva, surgical treatment was chosen by means of the inferior labial frenectomy. After the 45-day period of the surgical procedure, the repair process was adequate, the patient did not report painful symptoms, there were no signs of infection and there was an improvement in the quality of attached gingiva. It can be concluded that, since well indicated through a correct diagnosis, the lip frenectomy can result in a good prognosis and improvement of the quality of the attached gingiva. (AU)

Clinical Comparison of the Subepithelial Connective Tissue versus Platelet-Rich Fibrin for the Multiple Gingival Recession Coverage on Anterior Teeth Using the Tunneling Technique.

The aim of this study was to evaluate and compare, clinically, the efficiency of the subepithelial connective tissue graft (SCTG) and platelet-rich fibrin (L-PRF) using the tunnel technique to cover the multiple gingival recessions on anterior teeth, in the same patient. Within the limits of this study, we conclude that both SCTG and L-PRF proved to be reliable options for the treatment of gingival recessions, efficiently supporting the biological and aesthetic demand, stimulating the periodontal tissues' health, and bringing reliable and highly predictable results.

Comparison of Two Screw-Retained Free Gingival Grafting Techniques.

Free gingival graft is a predictable technique for increasing the amount of attached gingiva and root coverage; however, its use is limited for cosmetic reasons. To overcome this issue, this study sought to compare 2 free gingival graft techniques that use oral screws to attach grafts. Free gingival graft was performed on teeth 44 to 46 using the traditional technique, while on the opposite side, on teeth 34 to 36, partly epithelialized free gingival grafts were performed. The partly epithelialized free gingival grafts were found to provide better cosmetic results relative to the completely epithelialized free gingival graft, and the use of stabilizing screws was found to be simple and effective.

Biomateriais utilizados em substituição ao enxerto autógeno de gengiva / Biomaterials used in replacement of gingival autograft

A utilização de biomateriais na tentativa de reparar ou substituir tecidos perdidos não é recente e evoluiu juntamente com os avanços da medicina regenerativa visando à maior qualidade de vida da população. Na Odontologia a compreensão dos aspectos biomiméticos envolvidos após implantação de biomateriais como substitutos de tecido mole é de suma importância na prática clínica, uma vez que o enxerto autógeno de gengiva, considerado padrão ouro, possui inúmeras limitações em relação à dimensão da área a ser reparada, tempo cirúrgico e dor pós-operatória na loja doadora. Esta revisão tem como objetivo reunir estudos que propõem a utilização de substitutos ao enxerto autógeno de palato, ressaltando suas aplicabilidades, contraindicações, disponibilidade no mercado, além de resultados clínicos.(AU)

The use of biomaterials to repair or replace lost tissues is not new and has evolved with advances in regenerative medicine aimed at higher quality of life of the population. In dentistry, the understanding of biomimetic aspects after implantation of biomaterials used as soft tissue replacements is significant in practice, since the autograft gum considered the gold standard has many limitations regarding the size of the area to berepaired surgical time and postoperative pain in the donor area.This review aims to bring together studies that propose substitutes to autograft palate, highlighting its applicability, contraindications, availability in the market, and clinical outcomes. (AU)

MMP1-1607 polymorphism increases the risk for periapical lesion development through the upregulation MMP-1 expression in association with pro-inflammatory milieu elements.

OBJECTIVE: In this study, we evaluated the association between the MMP1-1607 polymorphism (rs1799750) and pro-inflammatory milieu elements with MMP-1 mRNA levels in vivo. MATERIAL AND METHODS: MMP1-1607 SNP and the mRNA levels of MMP-1, TNF-a, IFN-g, IL-17A, IL-21, IL-10, IL-4, IL-9, and FOXp3 were determined via RealTimePCR in DNA/RNA samples from patients presenting periapical granulomas (N=111, for both genotyping and expression analysis) and control subjects (N=214 for genotyping and N=26 for expression analysis). The Shapiro-Wilk, Fisher, Pearson, Chi-square ordinal least squares regression tests were used for data analysis (p<0.05 was considered statistically significant). RESULTS: The MMP1-1607 1G/2G and 1G/2G+2G/2G genotypes were significantly more prevalent in the patients than in controls, comprising a risk factor for periapical lesions development. MMP-1 mRNA levels were higher in periapical lesions than in healthy periodontal ligament samples, as well as higher in active than in inactive lesions. The polymorphic allele 2G carriers presented a significantly higher MMP-1 mRNA expression when compared with the 1G/1G genotype group. The ordered logistic regression demonstrated a significant correlation between the genetic polymorphism and the expression levels of MMP-1. Additionally, the pro- and anti-inflammatory cytokines IL-17A, IFN-g, TNF-a, IL-21, IL-10, IL-9, and IL-4 were significant as complementary explanatory variables of MMP-1 expression. CONCLUSION: The MMP1-1607 SNP was identified as a risk factor for periapical lesions development, possibly due to its association with increased MMP-1 mRNA levels in periapical lesions. The MMP-1 expression is also under the control of the inflammatory milieu elements, being the cytokines TNF-a, IL-21, IL-17A, and IFN-g associated with increased MMP-1 levels in periapical lesions, while IL-10, IL-9, or IL-4 presented an inverse association.

MMP1-1607 polymorphism increases the risk for periapical lesion development through the upregulation MMP-1 expression in association with pro-inflammatory milieu elements

ABSTRACT Increased matrix metalloproteinases (MMPs) activity is a hallmark of periapical granulomas. However, the factors underlying the MMPs expression modulation in healthy and diseased periapical tissues remains to be determined. Objective In this study, we evaluated the association between the MMP1-1607 polymorphism (rs1799750) and pro-inflammatory milieu elements with MMP-1 mRNA levels in vivo. Material and Methods MMP1-1607 SNP and the mRNA levels of MMP-1, TNF-a, IFN-g, IL-17A, IL-21, IL-10, IL-4, IL-9, and FOXp3 were determined via RealTimePCR in DNA/RNA samples from patients presenting periapical granulomas (N=111, for both genotyping and expression analysis) and control subjects (N=214 for genotyping and N=26 for expression analysis). The Shapiro-Wilk, Fisher, Pearson, Chi-square ordinal least squares regression tests were used for data analysis (p<0.05 was considered statistically significant). Results The MMP1-1607 1G/2G and 1G/2G+2G/2G genotypes were significantly more prevalent in the patients than in controls, comprising a risk factor for periapical lesions development. MMP-1 mRNA levels were higher in periapical lesions than in healthy periodontal ligament samples, as well as higher in active than in inactive lesions. The polymorphic allele 2G carriers presented a significantly higher MMP-1 mRNA expression when compared with the 1G/1G genotype group. The ordered logistic regression demonstrated a significant correlation between the genetic polymorphism and the expression levels of MMP-1. Additionally, the pro- and anti-inflammatory cytokines IL-17A, IFN-g, TNF-a, IL-21, IL-10, IL-9, and IL-4 were significant as complementary explanatory variables of MMP-1 expression. Conclusion The MMP1-1607 SNP was identified as a risk factor for periapical lesions development, possibly due to its association with increased MMP-1 mRNA levels in periapical lesions. The MMP-1 expression is also under the control of the inflammatory milieu elements, being the cytokines TNF-a, IL-21, IL-17A, and IFN-g associated with increased MMP-1 levels in periapical lesions, while IL-10, IL-9, or IL-4 presented an inverse association.

Effect of gingivitis treatment on the breath of chronic periodontitis patients.

PURPOSE: The present study assessed halitosis after treatment of gingivitis by using different diagnostic modalities. MATERIALS AND METHODS: A total of 27 patients (47 ± 7 years old; 14 women, 13 men) diagnosed with chronic periodontitis underwent supragingival mechanical debridement (day 0). Measurement of volatile sulfur compounds (VSC), visual analogue scale (VAS) and organoleptic assessment, visible plaque index (VPI) and gingival bleeding index (GBI) were obtained at baseline (prior to treatment of gingivitis) and then after 30, 90, and 180 days. The Friedman test was used to compare outcome data at days 0, 30, 90, and 180. Post-hoc comparison (tongue cleaning and non-cleaning) was performed using the Student t test for VSC and VAS and the Mann-Whitney test for organoleptic measurements. The level of significance was set at 5%. RESULTS: VSC and VAS means (days 0 and 180) were, respectively, 463.41 ppb (± 496.12 ppb) and 245.96 ppb (± 301.51 ppb) and 6.28 cm (± 2.23 cm) and 5.03 cm (± 2.01 cm). The frequency of high organoleptic scores (3 to 5) declined from 96.29% (day 0) to 81.48% (day 30), with a reduction of around 50% at day 180. Post-hoc comparison did not reveal any significant differences between the indicators analysed. CONCLUSION: Supragingival plaque control reduced halitosis in patients with periodontitis, but tongue cleaning vs no tongue cleaning did not yield different results.

Non-inflammatory destructive periodontal disease: a clinical, microbiological, immunological and genetic investigation.

Periodontitis comprises a group of multifactorial diseases in which periodontopathogens accumulate in dental plaque and trigger host chronic inflammatory and immune responses against periodontal structures, which are determinant to the disease outcome. Although unusual cases of non-inflammatory destructive periodontal disease (NIDPD) are described, their pathogenesis remains unknown. A unique NIDPD case was investigated by clinical, microbiological, immunological and genetic tools. The patient, a non-smoking dental surgeon with excessive oral hygiene practice, presented a generalized bone resorption and tooth mobility, but not gingival inflammation or occlusion problems. No hematological, immunological or endocrine alterations were found. No periodontopathogens (A. actinomycetemcomitans, P. gingivalis, F. nucleatum and T. denticola) or viruses (HCMV, EBV-1 and HSV-1) were detected, along with levels of IL-1ß and TNF-a in GCF compatible with healthy tissues. Conversely ALP, ACP and RANKL GCF levels were similar to diseased periodontal sites. Genetic investigation demonstrated that the patient carried some SNPs, as well HLA-DR4 (*0404) and HLA-B27 alleles, considered risk factors for bone loss. Then, a less vigorous and diminished frequency of toothbrushing was recommended to the patient, resulting in the arrest of alveolar bone loss, associated with the return of ALP, ACP and RANKL in GCF to normality levels. In conclusion, the unusual case presented here is compatible with the previous description of NIDPD, and the results that a possible combination of excessive force and frequency of mechanical stimulation with a potentially bone loss prone genotype could result in the alveolar bone loss seen in NIDPD.

Non-inflammatory destructive periodontal disease: a clinical, microbiological, immunological and genetic investigation

Periodontitis comprises a group of multifactorial diseases in which periodontopathogens accumulate in dental plaque and trigger host chronic inflammatory and immune responses against periodontal structures, which are determinant to the disease outcome. Although unusual cases of non-inflammatory destructive periodontal disease (NIDPD) are described, their pathogenesis remains unknown. A unique NIDPD case was investigated by clinical, microbiological, immunological and genetic tools. The patient, a non-smoking dental surgeon with excessive oral hygiene practice, presented a generalized bone resorption and tooth mobility, but not gingival inflammation or occlusion problems. No hematological, immunological or endocrine alterations were found. No periodontopathogens (A. actinomycetemcomitans, P. gingivalis, F. nucleatum and T. denticola) or viruses (HCMV, EBV-1 and HSV-1) were detected, along with levels of IL-1β and TNF-a in GCF compatible with healthy tissues. Conversely ALP, ACP and RANKL GCF levels were similar to diseased periodontal sites. Genetic investigation demonstrated that the patient carried some SNPs, as well HLA-DR4 (*0404) and HLA-B27 alleles, considered risk factors for bone loss. Then, a less vigorous and diminished frequency of toothbrushing was recommended to the patient, resulting in the arrest of alveolar bone loss, associated with the return of ALP, ACP and RANKL in GCF to normality levels. In conclusion, the unusual case presented here is compatible with the previous description of NIDPD, and the results that a possible combination of excessive force and frequency of mechanical stimulation with a potentially bone loss prone genotype could result in the alveolar bone loss seen in NIDPD.

Expression analysis of wound healing genes in human periapical granulomas of progressive and stable nature.

INTRODUCTION: Wound healing process involves the activation of extracellular matrix components, remodeling enzymes, cellular adhesion molecules, growth factors, cytokines and chemokines genes. However, the molecular patterns underlying the healing process at the periapical environment remain unclear. Here we hypothesized that endodontic infection might result in an imbalance in the expression of wound healing genes involved in the pathogenesis of periapical lesions. Furthermore, we suggest that differential expression of wound healing markers in active and latent granulomas could account for different clinical outcomes for such lesions. METHODS: Study samples consisted of 93 periapical granulomas collected after endodontic surgeries and 24 healthy periodontal ligament tissues collected from premolars extracted for orthodontic purposes as control samples. Of these, 10 periapical granulomas and 5 healthy periapical tissues were used for expression analysis of 84 wound healing genes by using a pathway-specific real-time polymerase chain reaction array. The remaining 83 granulomas and all 24 control specimens were used to validate the obtained array data by real-time polymerase chain reaction. Observed variations in expression of wound healing genes were analyzed according to the classification of periapical granulomas as active/progressive versus inactive/stable (as determined by receptor activator for nuclear factor kappa B ligand/osteoprotegerin expression ratio). RESULTS: We observed a marked increase of 5-fold or greater in SERPINE1, TIMP1, COL1A1, COL5A1, VTN, CTGF, FGF7, TGFB1, TNF, CXCL11, ITGA4, and ITGA5 genes in the periapical granulomas when compared with control samples. SERPINE1, TIMP1, COL1A1, TGFB1, and ITGA4 mRNA expression was significantly higher in inactive compared with active periapical granulomas (P < .001), whereas TNF and CXCL11 mRNA expression was higher in active lesions (P < .001). CONCLUSIONS: The identification of novel gene targets that curb the progression status of periapical lesions might contribute to a more accurate diagnosis and lead to treatment modalities more conducive to endodontic success.

MIF induces osteoclast differentiation and contributes to progression of periodontal disease in mice.

Periodontal disease (PD) is a chronic inflammatory and alveolar bone destructive disease triggered by microorganisms from the oral biofilm. Oral inoculation of mice with the periodontopathogen Aggregatibacter actinomycetemcomitans (Aa) induces marked alveolar bone loss and local production of inflammatory mediators, including Macrophage Migration Inhibitory Factor (MIF). The role of MIF for alveolar bone resorption during PD is not known. In the present study, experimental PD was induced in BALB/c wild-type mice (WT) and MIF knockout mice (MIF⁻/⁻) through oral inoculation of Aa. Despite enhanced number of bacteria, MIF⁻/⁻ mice had reduced infiltration of TRAP-positive cells and reduced alveolar bone loss. This was associated with decreased neutrophil accumulation and increased levels of IL-10 in periodontal tissues. TNF-α production was similar in both groups. In vitro, LPS from Aa enhanced osteoclastic activity in a MIF-dependent manner. In conclusion, MIF has role in controlling bacterial growth in the context of PD but contributes more significantly to the progression of bone loss during PD by directly affecting differentiation and activity of osteoclasts.

Mecanismos envolvidos na resposta imune e inflamatória frente à implantação de membrana de cortical óssea bovina no tecido subcutâneo de camundongos: caracterização histomorfométrica, imunoenzimática e molecular / Mechanisms involved in immune and inflammatory response to implantation of bovine bone cortical membrane in subcutaneous tissue of mice: characterization histomorphometric, imunoenzimatic and molecular

Os avanços relacionados à ciência dos biomateriais e engenharia tecidual buscam esclarecer os mecanismos envolvidos na resposta biológica associada ao uso desses dispositivos e sua interação com o sistema imune. Neste estudo, avaliou-se a resposta imune e inflamatória desenvolvida em camundongos frente à implantação de membrana de cortical óssea bovina no tecido subcutâneo, em implantação única e sequencial de duas membranas, assim como os possíveis mecanismos envolvidos no processo de reconhecimento e reabsorção desse biomaterial, de acordo com análise histomorfométrica, enzimática e molecular. Após a implantação da membrana, sinais de reabsorção que antes eram notados em pontos isolados, aos poucos se unem até sua completa degradação, observada somente após 15 dias. Todo o processo de reabsorção da membrana é acompanhado por uma reação inflamatória de magnitude moderada, seguida pelo declínio do número de leucócitos, surgimento de células gigantes multinucleadas e formação de uma cápsula de tecido conjuntivo fibroso. A cinética de TNF-α e MMP-2, MMP-8, MMP-9 e MMP-13 apresentou um padrão de produção decrescente, entretanto os níveis dos inibidores de metaloproteinases (TIMPs) e TGF-β parecem atuar de forma inversa. A velocidade de reabsorção após duas implantações consecutivas da membrana foi maior quando comparada ao grupo de animais que sofreu apenas uma implantação, porém os resultados do teste de hipersensibilidade do tipo tardia (DTH) demonstraram que a membrana é biocompatível, pois não elicitou resposta imunológica exacerbada após uma segunda implantação, confirmando então a natureza não imunogênica desse biomaterial. Finalmente, os animais CD14KO e MyD88KO apresentaram uma reabsorção mais lenta da membrana implantada, quando comparados aos animais C57Bl/6 (WT)...

Advances related to the biomaterials science and tissue engineering seek to clarify the mechanisms involved in the biological response associated with the use of those devices and their interaction with the immune system. This study evaluated the inflammatory and immune response developed in mice after implantation bovine bone cortical membrane in subcutaneous tissue, in both, unique and sequential implantation of 2 membranes, as the mechanisms involved in this biomaterial recognition and resorption process, on regards to histomorphometric, enzymatic and molecular analysis. After membrane implantation, previously observed signs of resorption in isolated spots, gradually unite until their complete degradation after 15 days. The whole membrane resorption process is accompanied by a moderate inflammatory reaction, followed by a decline in the leukocytes number, appearance of multinucleated giant cells and formation of a capsule of fibrous connective tissue. The kinetics of TNF-α, MMP-2, MMP-8, MMP-9 e MMP-13 showed a pattern of decreasing production, however, levels of metalloproteinases inhibitors (TIMPs) and TGF-β seem to act in reverse way. The resorption rate after two successive membrane implantations was higher when compared to the group which suffered only one implantation, however, the results of delayed test hypersensity (DTH) demonstrated that the membrane is biocompatible, that is, it does not elicited too high immune response after a second position, confirming the non immunogenic nature of this biomaterial. Eventually, CD14KO and MyD88KO strains showed a slower membrane resorption when compared to animals C57Bl/6, demonstrating that the CD14 and MyD88 molecules are involved in biomaterial recognition and play an important role in bovine cortical bone membrane resorption process, indicating that PAMPs and/or DAMPs are involved in biological response generated by this biomaterial.

Dose-response met-RANTES treatment of experimental periodontitis: a narrow edge between the disease severity attenuation and infection control.

Chemokines and chemokine receptors have been implicated in the selective migration of leukocyte subsets to periodontal tissues, which consequently influences the disease outcome. Among these chemoattractants, the chemokines CCL3, CCL4 and CCL5 and its receptors, CCR1 and CCR5, have been associated with increased disease severity in mice and humans. Therefore, in this study we investigated the modulation of experimental periodontitis outcome by the treatment with a specific antagonist of CCR1 and 5 receptors, called met-RANTES. C57Bl/6 mice was orally infected with Aggregatibacter actinomycetemcomitans and treated with 0.05, 0.1, 0.5, 1.5 and 5 mg doses of met-RANTES on alternate days, and evaluated by morphometric, cellular, enzymatic and molecular methods. At 0.5 mg up to 5 mg doses, a strong reduction in the alveolar bone loss and inflammatory cell migration were observed. Interestingly, 5 mg dose treatment resulted in the maximum inhibition of inflammatory cell migration, but resulted in a similar inhibition of bone loss when compared with the lower doses, and also resulted in increased bacterial load and CRP response. When 0.5 and 5 mg therapy regimens were compared it was observed that both therapeutic protocols were able to downregulate the levels of pro-inflammatory, Th1-type and osteoclastogenic cytokines, and CD3+ and F4/80+ cells migration to periodontal tissues, but the high dose modulates host response in a more pronounced and unspecific and excessive way, interfering also with the production of antimicrobial mediators such as MPO, iNOS and IgG, and with GR1+ and CD19+ cells migration. Our results demonstrate a thin line between beneficial immunoregulation and impaired host defense during experimental periodontitis, and the determination of the exact equilibrium point is mandatory for the improvement of immune-targeted therapy of periodontitis.

Evidences of the cooperative role of the chemokines CCL3, CCL4 and CCL5 and its receptors CCR1+ and CCR5+ in RANKL+ cell migration throughout experimental periodontitis in mice.

Periodontal disease (PD) is characterized by the inflammatory bone resorption in response to the bacterial challenge, in a host response that involves a series of chemokines supposed to control cell influx into periodontal tissues and determine disease outcome. In this study, we investigated the role of chemokines and its receptors in the immunoregulation of experimental PD in mice. Aggregatibacter actinomycetemcomitans-infected C57Bl/6 (WT) mice developed an intense inflammatory reaction and severe alveolar bone resorption, associated with a high expression of CCL3 and the migration of CCR5+, CCR1+ and RANKL+ cells to periodontal tissues. However, CCL3KO-infected mice developed a similar disease phenotype than WT strain, characterized by the similar expression of cytokines (TNF-alpha, IFN-gamma and IL-10), osteoclastogenic factors (RANKL and OPG) and MMPs (MMP-1, MMP-2, MMP-3, TIMP-1 and TIMP-3), and similar patterns of CCR1+, CCR5+ and RANKL+ cell migration. The apparent lack of function for CCL3 is possible due the relative redundancy of chemokine system, since chemokines such as CCL4 and CCL5, which share the receptors CCR1 and CCR5 with CCL3, present a similar kinetics of expression than CCL3. Accordingly, CCL4 and CCL5 kinetics of expression after experimental periodontal infection remain unaltered regardless the presence/absence of CCL3. Conversely, the individual absence of CCR1 and CCR5 resulted in a decrease of leukocyte infiltration and alveolar bone loss. When CCR1 and CCR5 were simultaneously inhibited by met-RANTES treatment a significantly more effective attenuation of periodontitis progression was verified, associated with lower values of bone loss and decreased counts of leukocytes in periodontal tissues. Our results suggest that the absence of CCL3 does not affect the development of experimental PD in mice, probably due to the presence of homologous chemokines CCL4 and CCL5 that overcome the absence of this chemokine. In addition, our data demonstrate that the absence of chemokine receptors CCR1+ and CCR5+ attenuate of inflammatory bone resorption. Finally, our data shows data the simultaneous blockade of CCR1 and CCR5 with MetRANTEs presents a more pronounced effect in the arrest of disease progression, demonstrating the cooperative role of such receptors in the inflammatory bone resorption process throughout experimental PD.