Use of enamel matrix- derived proteins combined with bone graft for the treatment of intrabony defects: overview of reviews (FRISBEE review) / Uso de proteínas derivadas de la matriz del esmalte combinado con injerto óseo para el tratamiento de defectos intraóseos: resumen estructurado de revisiones sistemáticas (revisión FRISBEE)

Introduction: The use of enamel matrix-derived proteins (EMD) has increased in recent years due to their tissue-inducing properties that support periodontal regeneration. This study is an overview of systematic reviews with FRISBEE methodology on the use of EMD alone or combined with autologous bone graft materials (BGM) in the treatment of intrabony defects. Materials and Methods: A systematic search in the Epistemonikos database was performed. RevMan 5.3 and GRADEpro were used for data analysis and presentation Results: Four systematic reviews and two clinical trials were identified. All studies analysed change in probing depth, clinical attachment level, gingival margin level and bone defect depth (all changes in favour of EMD+BGM groups: mean difference (MD): 0.37 mm more, MD: 0.7 mm more, MD: 0.3 mm less, MD: 0.75 more, respectively). Conclusions: Adding autologous bone graft to EMD to treat intrabony defects showed better results, but not a relevant clinical difference compared to the use of EMD alone.

Introducción: El uso de proteínas derivadas de la matriz del esmalte (EMD) ha aumentado en los últimos años debido a sus propiedades inductoras de tejidos que apoyan la regeneración periodontal. Este estudio es una revisión sistemática de revisiones sistemáticas utilizando metodología FRISBEE sobre el uso de EMD solo o combinado con materiales injerto óseo autólogo (BGM) en el tratamiento de defectos intraóseos. Materiales y Métodos: Se realizó una búsqueda sistemática en la base de datos Epistemonikos. Se utilizaron RevMan 5.3 y GRADEpro para el análisis y la presentación de los datos. Resultados: Se identificaron cuatro revisiones sistemáticas y dos ensayos clínicos. Todos los estudios analizaron el cambio en la profundidad de sondaje, el nivel de inserción clínica, el nivel del margen gingival y la profundidad del defecto óseo (todos los cambios a favor de los grupos EMD+BGM: MD: 0,37 mm más, media de diferencia (MD): 0,7 mm más, MD: 0,3 mm menos, MD: 0,75 más, respectivamente). Conclusión: La adición de injerto óseo autólogo a la EMD para tratar defectos intraóseos mostró mejores resultados, pero no una diferencia clínica relevante en comparación con el uso de la EMD sola.

Avaliação radiográfica da resposta pulpar e periapical de dentes de cães ao Emdogain® após pulpotomia / Radiographic evaluation of pulpal and periapical response of dogs teeth to Emdogain® used as pulpotomy agent

Introdução: O derivado da matriz do esmalte (Emdogain®) é um extrato de proteína usado para cicatrização periodontal. Objetivo: O objetivo desse estudo foi avaliar radiograficamente a resposta pulpar e periapical de dentes de cães após pulpotomia e uso do gel derivado da matriz do esmalte (Emdogain®). Métodos: A pulpotomia foi realizada em 30 dentes (60 raízes) de 3 cães, e o tecido pulpar remanescente foi recoberto com os seguintes materiais: Grupos 1 e 4: gel derivado da matriz do esmalte (Emdogain®); Grupos 2 e 5: hidróxido de cálcio; Grupos 3 e 6: óxido de zinco e cimento de eugenol. Após 10 dias (Grupos 1-3) e 75 dias (Grupos 4-6) foram obtidas radiografias periapicais e a avaliação radiográfica foi realizada considerando-se: a integridade da lâmina dura, presença de áreas de rarefação óssea periapical, reabsorção radicular (interna e externa) e formação de ponte de dentina. Resultados: No período de 10 dias, todos os espécimes dos Grupos 1-3 apresentaram ausência de rarefação óssea periapical, reabsorção radicular (interna e externa) e formação de ponte dentinária. No período de 75 dias, os Grupos 4-6 não apresentaram formação de ponte dentinária em nenhum espécime. Áreas de rarefação óssea periapical foram observadas em 100% das raízes no Grupo 4, 62,5% das raízes no Grupo 6 e em 25% das raízes nos Grupos 5. Conclusão: O uso do gel derivado da matriz do esmalte (Emdogain®) como material para capeamento após a pulpotomia levou à formação de lesões periapicais e não induziu a deposição de tecido mineralizado (AU).

Introduction: The enamel matrix derivative (Emdogain®) is a protein extract used for periodontal healing. The objective of this study was to evaluate radiographically the pulpal and periapical response of dogs teeth after pulpotomy and use of enamel matrix derivative gel (Emdogain®). Methods: Pulpotomy was performed in 30 teeth (60 roots) from 3 dogs and the remaining pulp tissue was capped with the following materials: Groups 1 and 4: enamel matrix derivative gel (Emdogain®); Groups 2 and 5: calcium hydroxide; Groups 3 and 6: zinc oxide and eugenol cement. After 10 days (Groups 1-3) and 75 days (Groups 4-6) periapical radiographs were obtained and the radiographic evaluation was performed considering the integrity of the lamina dura, presence of areas of periapical bone rarefaction, root resorption (internal and external) and dentin bridge formation. Results: In the 10- day period, all specimens in Groups 1-3 presented absence of periapical bone rarefaction, absence of root resorption (internal and external) and absence of dentin bridge formation. In the 75-day period, Groups 4-6 did not present dentin bridge formation in any specimen. Periapical bone rarefaction areas were observed in 100% of the roots in Group 4, 62,5% of the roots in Group 6 and in 25% of the roots in Groups 5. Conclusion: The use of enamel matrix derivative gel (Emdogain®) as a capping material after pulpotomy lead to formation of periapical lesions and did not induce deposition of mineralized tissue(AU).

Three-dimensional assessment of enamel and dentine in mouse molar teeth during masseter muscle hypofunction / Evaluación tridimensional del esmalte y la dentina de dientes molares de ratones durante la hipofunción del músculo masetero

Background: Mouse molar is a widely used model for teeth development. However, the effect of masticatory function on enamel and dentine in adult individuals remains poorly understood. As reported, the unilateral masseter hypofunction induced by botulinum toxin type A (BoNTA) resulted in mandibular bone damage and signs of unilateral chewing in adult mice. Objective: We aimed to assess the amount of enamel and dentine in the first molar (M1) during the unilateral masseter hypofunction in mice, using high-resolution X-ray microtomography (µCT) as threedimensional approach. Materials and methods: Mandibles of adult BALB/c mice, located either in a Control-group (without intervention) or a BoNTA-group, were ex-vivo scanned using µCT. Treated individuals received each one BoNTA intervention in the right masseter, and saline solution in the left masseter (intra-individual control). Enamel and dentine from M1 were segmented, and volume, thickness and mesial root length were quantified. Results: Enamel volume from treated side resulted unchanged after 2 weeks of unilateral masseter hypofunction. No differences for enamel volume were found between both sides of control individuals, and between these and samples from hypofunctional side in BoNTA-group. Enamel volume from saline-injected side was reduced when compared with experimental side (p<0,01). No differences in dentine volume, thickness of enamel and dentine, and mesial root length were found for any group. Conclusion: The amount of enamel in hypofunctional molars remains unaffected after unilateral BoNTA intervention in the masseter, but contralateral side showed reduced enamel volume. Therefore, increased functional wearing during unilateral chewing after BoNTA intervention should be considered.

Introducción: El molar de ratón es utilizado como modelo de estudio en el desarrollo dental. El efecto de la función masticatoriasobre el tejido dental en individuos adultos aún se comprende. En ratones adultos, la hipofunción unilateral del masetero inducida por toxina botulínica tipo A (BoNTA) resultó en daño óseo mandibular y signos de masticación unilateral. Objetivo: Evaluamos la cantidad de esmalte y dentina en el primer molar (M1) durante la hipofunción unilateral del músculo masetero en ratones mediante análisis con microtomografía (µCT). Materiales y métodos: Las mandíbulas de ratones BALB/c adultos, del grupo Control (sin intervención) o el grupo BoNTA, fueron escaneadas ex-vivo con µCT. Los individuos tratados se inyectaron con BoNTA en el masetero derecho y con solución salina en el masetero izquierdo (control intra-individuo). El volumen y grosor de esmalte y dentina del M1, y la longitud de la raíz mesial fueron medidos. Resultados: No hubo cambios en el volumen del esmalte del lado tratado con BoNTA y en ambos lados del grupo Control, 2 semanas post-intervención. El esmalte del lado control intra-individuo se redujo comparado con el lado experimental (p< 0,01). No hubo cambios en el volumen de dentina, el grosor de esmalte y dentina o en longitud de la raíz mesial de ambos grupos. Conclusión: La cantidad de esmalte en los molares hipofuncionales no se afecta después de la inyección unilateral de BoNTA en masetero, pero si se reduce en el lado contralateral. Por lo tanto, se debe considerar un desgaste dental asimétrico durante esta intervención.

Genes involved in amelogenesis imperfecta. part i / Genes involucrados en la amelogénesis imperfecta. parte i

ABSTRACT Amelogenesis imperfecta (AI) refers to a group of genetic alterations of the normal structure of the dental enamel that disturbs its clinical appearance. AI is classified as hypoplastic, hypocalcified, and hypomaturation. These abnormalities may exist in isolation or associated with other systemic conditions in the context of a syndrome. This article is aimed to thoroughly describe the genes involved in non-syndromic AI, the proteins encoded by these genes and their functions according to current scientific evidence. An electronic literature search was carried out from the year 2000 to December of 2017, preselecting 1,573 articles, 63 of which were analyzed and discussed. The results indicated that mutations in 16 genes are responsible for non-syndromic AI: AMELX, AMBN, ENAM, LAMB3, LAMA3, ACPT, FAM83H, C4ORF26, SLC24A4, ITGB6, AMTN, MMP20, KLK4, WDR72, STIM1, GPR68. Future research with a translational approach will help to identify new mutations or genes, contributing to the evolution in the way of classifying, diagnosing and treating the various types of amelogenesis imperfecta.

RESUMEN La amelogénesis imperfecta (AI) constituye un grupo de alteraciones de la estructura normal del esmalte dental de origen genético que perturba su apariencia clínica. La AI se clasifica en hipoplásica, hipomadura e hipocalcificada. Estas anomalías pueden existir de manera aislada o asociada a otras afecciones sistémicas en el marco de un síndrome. En el presente artículo se pretende describir de manera detallada los genes involucrados en la AI no sindrómica, las proteínas codificas por estos genes y sus funciones, de acuerdo a la evidencia científica actual. Se realizó una búsqueda electrónica de literatura desde el año 2000 hasta diciembre de 2017, haciendo una preselección de 1573 artículos, de los cuales 63 fueron analizados y discutidos. Los resultados indicaron que las mutaciones en 16 genes son responsables de la AI no sindrómica: AMELX, AMBN, ENAM, LAMB3, LAMA3, ACPT, FAM83H, C4ORF26, SLC24A4, ITGB6, AMTN, MMP20, KLK4, WDR72, STIM1, GPR68. Las futuras investigaciones abordadas desde la visión translacional ayudarán a identificar nuevas mutaciones o nuevos genes, lo cual contribuirá a la evolución en la manera de clasificar, diagnosticar y tratar los diferentes tipos de amelogénesis imperfecta.

Osteogain: Um novo biomodificador para estímulo ósseo? / Osteogain: a novel biomodifier to stimulate bone formation?

Objetivo: avaliar, a partir de revisão de literatura, o uso de matriz derivada de esmalte na forma líquida (Osteogain) junto com biomateriais, para aprimorar a regeneração em destaque na formação óssea. Material e métodos: realizou-se busca de artigos através do PubMed e outras bases de dados eletrônicas no Medline, até o mês maio de 2017. Utilizou os seguintes MeSH terms "Osteogain" OR "enamel matrix proteins liquid". Dos artigos selecionados criou-se uma tabela de resumo. Resultados: dos 18 artigos encontrados, oito artigos foram selecionados e separados para leitura completa. Houve apenas dois estudos in vivo, uma revisão breve do Osteogain e cinco análises in vitro. Toda a literatura mostrou-se favorável ao uso do Osteogain. A nova formulação líquida de matriz derivada de esmalte mostrou induzir a mineralização óssea e foi positiva quanto à fixação celular nas partículas de enxerto ósseo, diferenciação/mineralização dos osteoblastos, expressão gênica de muitas citocinas e fatores de crescimento. Conclusão: o Osteogain é uma alternativa biológica favorável para estímulo ósseo, com melhor adsorção de proteína nos materiais de enxerto ósseo, sendo uma escolha promissora na regeneração óssea, principalmente em defeitos complexos.

Objective: to evaluate from a literature review the use of Enamel Matrix Derivative Liquid (Osteogain) in combination with biomaterials, to improve tissue regeneration, featured in bone formation. Material and methods: a literature search was performed on PubMed and others Medline electronic databases until May 2017, using the following MeSH terms "Osteogain" OR "liquid enamel matrix proteins". From the selected articles a summary table has been created. Results: from the 18 articles, 8 were selected and separated for full-text screening. There were only two in vivo studies, one brief review about Osteogain and five in vitro analyzes. All available literature was favorable to the use of Osteogain. The new liquid formulation of enamel matrix derivative showed to induce bone mineralization and was positive effect on fi xing cell bone graft particles, differentiation/mineralization of osteoblasts, gene expression of many cytokines and growth factors. Conclusion: osteogain is presented as a promising biological alternative for bone regeneration, with superior protein adsorption to bone graft materials, being considered a potential choice for tissue regeneration, especially in non-contained defects.

Tratamento das recessões gengivais utilizando o enxerto de tecido conjuntivo isolado e associado às proteínas derivadas da matriz do esmalte ­ uma revisão integrativa / Gingival recession treatment using subepithelial connective tissue graft isolated and associated to enamel matrix protein ­ an integrative review

Objetivos: A utilização do enxerto de tecido conjuntivo subepitelial é considerada o tratamento padrão-ouro para o recobrimento das recessões gengivais classe I e II de Miller. Porém, alguns fatores como a necessidade de um segundo sítio cirúrgico para a obtenção do enxerto, a limitada quantidade de tecido a ser doado, um maior tempo cirúrgico, e um possível desconforto pós-operatória, tem incentivado pesquisas utilizando biomateriais. Em função disso, o objetivo desta revisão integrativa foi realizar uma comparação entre os tratamentos da recessão gengival utilizando o enxerto de tecido conjuntivo isoladamente, e quando associado às proteínas derivadas da matriz do esmalte, buscando avaliar o benefício da sua utilização em relação aos parâmetros de profundidade de sondagem, nível de inserção clínica, espessura e largura do tecido queratinizado, cobertura de raiz e estabilidade da cobertura radicular ao longo do tempo. Material e métodos: Dois revisores de forma independente realizaram a pesquisa em bases de dados eletrônicas, entre os anos de 2000 e 2016, buscando ensaios clínicos randomizados em humanos. Foram encontrados 266 artigos no total, que após a leitura de título e resumo, e leitura na íntegra, foram selecionados para compor esta revisão 3 artigos. Resultados: Não foram observadas diferenças estatisticamente significativas entre o tratamento com o enxerto de tecido conjuntivo subepitelial isolado e em associação com o Emdogain®, para os parâmetros apresentados. Conclusão: Apesar de ambos os tratamentos apresentarem resultados clínicos satisfatórios, não se apresentam benefícios adicionais da associação do enxerto de tecido conjuntivo com o Emdogain®. (AU)

Objective: The use of subepithelial connective tissue graft is considered the gold standard treatment for the coverage of gingival recessions Miller's Class I and II. However, some considerations as the need of a second surgical procedure to obtain the graft, a limited amount of tissue that can be donated, a longer surgical duration, and a possible post-operative discomfort have improved researches using biomaterials. According to that, the objective of this integrative review was to compare treatments to gingival recession using only subepithelial connective tissue grafts and grafts associated to enamel matrix protein, aiming to evaluate the benefits of its usage considering the probing depths parameters, clinical insertion levels, thickness and width of keratinized tissue, root coverage and its stability through time. Material and methods: Two reviewers searched independently in electronic databases for clinical trials in humans from 2000 to 2016 that were related to this approach. It was found initially 266 articles in total. After title, abstract and full-text reading, three articles were selected to compose this review. Results: It was not observed statistically significant difference between the subepithelial connective tissue graft isolated and associated with Emdogain®. Conclusion: Even though both treatments showed satisfactory clinical results, there is no additional benefit associating the graft and Emdogain®

Pathogenesis of dental fluorosis: biochemical and cellular mechanisms / Patogénesis de la fluorosis dental: mecanismos bioquímicos y celulares

Abstract. The mechanisms involved in the development of dental fluorosis are still unknown. The development of in vivo and in vitro models using biologically relevant concentrations of fluoride for the emergence of fluorosis has allowed suggesting hypotheses that contribute to the understanding of the mechanisms that produce this defect in enamel development, with high prevalence in Colombia. This topic review presents an update on the normal mechanisms of the formation of enamel and how they are affected by exposure to high concentrations of fluoride. This is a thorough review of the deleterious effects of fluoride on the cells and the extracellular matrix, especially during the maturation stage, resulting in a delay of the removal of the protein matrix of amelogenins, as well as the appearance of mottled enamel-a characteristic of dental fluorosis. Finally, it shows the perspectives of the study of this defect in enamel development from biochemistry and cellular and molecular biology.

RESUMEN. Los mecanismos involucrados en el desarrollo de la fluorosis dental aún no se conocen a cabalidad. El desarrollo de modelos in vivo e in vitro que utilizan concentraciones de fluoruro biológicamente relevantes para la aparición de fluorosis ha permitido el planteamiento de hipótesis que aportan cada vez más al conocimiento de los mecanismos que generan este defecto del desarrollo del esmalte, de alta prevalencia en Colombia. Esta revisión presenta una actualización sobre los mecanismos normales de la formación del esmalte y cómo estos se ven afectados por la exposición a altas concentraciones de fluoruro. Se presenta una revisión en detalle de los efectos deletéreos del fluoruro sobre las células y sobre la matriz extracelular, especialmente durante la etapa de maduración, que tendrán como consecuencia el retraso de la remoción de la matriz proteica de amelogeninas y se traducirá en la apariencia de esmalte moteado, característica de la fluorosis dental. Por último, se muestran las perspectivas del estudio de este defecto del desarrollo del esmalte desde la bioquímica y la biología celular y molecular.

Uso da matriz derivada do esmalte (mde) associada ao vidro bioativo no tratamento de defeitos infra-ósseos: uma revisão de literatura / Use of enamel matrix derivate (EMD) associated with bioactive glass in the treatment of intraosseous defects: A Literature Review

A regeneração periodontal tem como objetivo recuperar as estruturas perdidas (osso alveolar, cemento e ligamento periodontal) como sequelas da doença periodontal. O conhecimento da Matriz Derivada do Esmalte (MDE) tem na literatura uma ampla aplicação, porém sua relação direta associada aos vidros bioativos é pouco caracterizada em todas suas vias, nesse contexto este estudo teve como proposta baseada numa revisão literária por meio de bases de dados (Pubmed, Scielo, lilacs), discutir aspectos importantes sobre a avaliação da efetividade clínica do vidro bioativo em combinação com a MDE (Emdogain) principalmente em defeitos infra-ósseos. O Emdogain (EMD) sozinho ou em associação a outros biomaterais de enxertia parece promover ganhos na regeneração tecidual de tecidos perdidos pela periodontite, porém sua associação tem-se poucos resultados significativos que justifiquem sua ampla utilização, sua indicação deve estar baseada em um bom diagnóstico periodontal e da morfologia do defeito infraósseo. Assim, tornam-se necessários mais estudos para elucidar interações e mecanismos celulares envolvidos no complexo de ação para justificar o seu uso.(AU)

Periodontal regeneration aims to recover the lost structures (alveolar bone, cementum and periodontal ligament) as sequelae of periodontal disease. The knowledge derived from the enamel matrix (EMD) is in the literature a wide application, but its direct relationship associated with bioactive glass is poorly characterized in all its way in this context this study was proposed based on a literature review through databases (Pubmed, Scielo, lilacs), discuss important aspects of the evaluation of the clinical effectiveness of bioactive glass in combination with EMD (Emdogain) mainly intraosseous defects.Emdogain (EMD) alone or in association with other biomaterials of grafting appears to promote gains in the tissue regeneration of lost tissues by periodontitis, but its association has few significant results that justify its wide use, its indication must be based on a good periodontal diagnosis and infraosseous defect morphology. Thus, further studies are needed to elucidate interactions and cellular mechanisms involved in the action complex to justify its use.(AU)

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. .

Effect of enamel matrix derivatives on the differentiation and proliferation of human periodontal ligament stem cells / 中国组织工程研究

BACKGROUND:The enamel matrix derivative has been used in the clinical treatment of severe periodontitis; however, the mechanism(s) by which enamel matrix derivative promotes periodontal regeneration is stil obscure. OBJECTIVE:To explore the effects of enamel matrix derivatives on the differentiation and proliferation of periodontal ligament stem cels. METHODS:Periodontal ligament stem cels were isolated and identified from human teeth. Cloning forming efficiency, surface antigen expression and pluripotency were detected and identified. Enamel matrix derivatives with different concentrations (20, 50, 100 mg/L) were used to culture periodontal ligament stem cels for 2 and 4 weeks. Colagen synthesis and mineralized nodule formation were detected using Trichrom staining and Von Kosa’s staining, respectively; real-time RT-PCR was employed to detect expressions of colagen type I, osteocalcin, and RUX2; MTT and cel growth rate assay were used to detect the proliferation of periodontal ligament stem cels. RESULTS AND CONCLUSION:Periodontal ligament stem cels were spindle-shaped and showed a higher colony forming efficiency than periodontal ligament cels. The expressions of surface antigens of periodontal ligament stem cels-CD105, CD29, CD45, CD44 were respectively 99.8%, 99.7%, 1.26%, 98.8%, indicating periodontal ligament stem cels have the multilineage differentiation potential. Enamel matrix derivatives improve the colagen synthesis and mineralization nodule formation of periodontal ligament stem cels in a time-dose dependent manner. They also can improve the expression of osteogensis-related genes colagen type I, osteocalcin, RUX2 and proliferation of periodontal ligament stem cels.

Do the Clinical Effects of Enamel Matrix Derivatives in Infrabony Defects Decrease Overtime? A Systematic Review and Meta-Analysis

Previous systematic reviews have demonstrated better results with enamel matrix derivative proteins (EMDP) as compared with open flap debridement (OFD) for the management of infrabony periodontal defects (IPD). The aim of this study was to determine whether these differences vary according to the follow-up and quality of the studies. Cochrane Central Register of Controlled Trials, Medline/PubMed, Lilacs, Embase and Web of Science electronic databases were searched up to August 2013 for randomized clinical trials.Eligible outcomes were changes in probing depth (PD), clinical attachment level (CAL),gingival recession (GR) and bone changes (BC). Studies with follow-up of 12 months showed differences of 0.97 mm (CI95% 0.52 - 1.43) and 1.19 mm (CI95% 0.77 - 1.60) for PD and CAL, respectively, favorable for EMDP. Studies with follow-up ≥ 24 months presented advantages of 1.11 mm (CI95% 0.74 -1.48) for CAL and 0.83 mm (CI95% 0.19 -1.48) for PD,with use of EMDP. Considering the quality of studies, those with low risk of bias showed lower difference between groups, presenting 0.8 mm (CI95% 0.24-1.36) for CAL, favorable for EMDP and without differences for PS (0.51 mm, CI95% -0.21 - 1.23). In conclusion, follow-up time (< or > 2 years) and the risk of bias influence the results of treatment with EMDP in IPD.

Revisões sistemáticas prévias tem demonstrado melhores resultados com proteínas derivadas da matriz de esmalte (PDME) em comparação a retalho de espessura total (RET) para o manejo de defeitos periodontais infraósseos (DPI). O objetivo desse estudo foi determinar se essas diferenças variam de acordo com o tempo de acompanhamento e com a qualidade dos estudos. As bases de dados Cochrane Central Register of Controlled Trials, MEDLINE (PubMed), Lilacs, Embase e Web of Science foram pesquisadas sem limitação de tempo ate agosto de 2013 para ensaios clínicos randomizados. Os desfechos elegíveis foram alterações na profundidade de sondagem (PS), nível de inserção clinica (NIC), recessão gengival (RG) e alterações ósseas (AO). Resultados: Estudos com acompanhamento de ate 12 meses mostraram diferenças de 0.97 mm (CI95% 0.52 – 1.43) e 1.19 mm (CI95% 0.77 – 1.60) para PS e NIC, favoráveis a PDME, respectivamente. Estudos com acompanhamento ≥24 meses demonstraram vantagens de 1.11 mm (CI95% 0.74 -1.48) para NIC e 0.83 mm (CI95% 0.19 -1.48) para PS, com o uso de PDME. Considerando a qualidade dos estudos, publicações com baixo risco de viés exibiram menores diferenças entre os grupos apresentando 0.8 mm (CI95% 0.24-1.36) para o NIC, sem diferenças para PS (0.51 mm, CI95% -0.21 – 1.23). Pode-se concluir que o tempo de acompanhamento (< ou > 2anos) e o risco de viés são variáveis que influenciam nos resultados do tratamento com PDME em DPIO.

Estandarizacion de un protocolo de extraccion y caracterizacion del material proteico del esmalte dental erupcionado / Standardization of a protocol of extraction and characterization of erupted dental enamel proteins

Introduccion: El 95% del peso del esmalte dental erupcionado sano corresponde a material inorganico, 4% agua y 1% a materia organica (con un 0,03 a 0,1% de trazas de proteinas estructurales). En el caso del esmalte con defectos del desarrollo - como la fluorosis dental - existe evidencia de la retencion de material proteico, despertando la necesidad del estudio del proteoma del esmalte en estas condiciones. Limitaciones inherentes a los tejidos duros, como su bajo contenido proteico encerrado en una matriz mineral, dificultan los procesos de extraccion y caracterizacion de proteinas. El presente estudio, busca hacer una adaptacion metodologica de los procedimientos de extraccion de proteinas en esmalte dental erupcionado humano, para su posible aplicacion al estudio del proteoma del esmalte con defectos del desarrollo. Objetivo: Estandarizar una tecnica de extraccion y caracterizacion del material proteico del esmalte de dientes erupcionados. Materiales y metodos: Se recolectaron dientes sanos con extraccion indicada y se realizaron: -cortes de secciones longitudinales de 550 ¦Ìm, - separacion mecanica de esmalte/dentina y - pulverizacion de esmalte dental. El pulverizado se sometio a desmineralizacion/precipitacion de proteinas con TCA 12%. El extracto fue separado por electroforesis SDS-PAGE y caracterizado por LC-MS/ MS. Resultados: el procedimiento fue estandarizado. No se evidenciaron bandas de proteina despu¨¦s de la electroforesis SDS-PAGE, pero se identificaron y caracterizaron 138 peptidos, correspondientes a 13 proteinas, 3 de ellas especificas del esmalte (Amelogenina X, Amelogenina Y, Ameloblastina). Conclusiones: por primera vez en Colombia, se estandarizan y se adaptan metodos de extraccion y caracterizacion de proteinas del esmalte dental, abriendo las puertas al estudio del proteoma de este tejido.

Introduction: 95% of erupted enamel corresponds to inorganic material, 1% organic and 4% water content (by weight) and it only has traces of structural proteins (0,03¨C0,1%), making extraction and characterization a difficult process. Enamel developmental defects as dental fluorosis, have been related to protein retention, indicating a clear need for the study of the enamel proteome. Standardization of methods for extraction and characterization of enamel proteins will allow the characterization of the human enamel proteome under these particular conditions. Methods: ethical approval from Universidad El Bosque ethics committee was granted and informed consent was given. Human permanent erupted teeth were collected from Universidad El Bosque Dental Clinics. The teeth crowns were cut in 550 ¦Ìm sections, and dental tissues were separated. The enamel sections were grinded with liquid nitrogen to get enamel powder. Powdered enamel was demineralized and the proteins precipitated with TCA 12%. The proteins were separated by SDS-PAGE Electrophoresis and characterized by LC-MS/MS. Results: enamel powdering and protein extraction techniques were standardized at Universidad El Bosque laboratories. No protein bands were detected after SDS- PAGE Electrophoresis, however, by LC-MS/MS, 138 peptides from 13 proteins were identified, 3 of them enamel-specific (Amelogenin X, Amelogenin Y). Conclusions: for the first time in Colombia, methods for extraction and characterization of proteins are standardized and applied on dental enamel, opening the doors for the study of the proteome from sound and defective dental enamel.

Immunohistochemical localization of enamelin in developing rat tooth germ / 北京大学学报(医学版)

Objective: To observe the immunohistochemical localization of enamelin in enamel formationand mineralization. Methods: Tissue sections of the first mandibular molar tooth germ from 1, 3, 7, 10, 14 days rats after birth were prepared, expression of the enamelin protein was identified by immunohistochemical technique. Results: Enamelin was found in the cytoplasm of ameloblasts in 1-10 days old rat postnatal first mandibular molar tooth germs. Enamelin appeared weakly in the tooth germs of 1 day rats. From 3 to 10 days, enamelin localized both in the cytoplasm of ameloblasts and the uncalcified enamel from the dentino-enamel junction to surfaces of the tooth. Enamelin protein was negative in the tooth germs of 14 days rats postnatally. Conclusion: Enamelin protein is synthesised and secreted by ameloblasts, specially localized in enamel from DEJ to surfaces of the tooth, suggesting that enamelin has important roles in enamel formation.

An in vivo study on osteoinductive property of enamel matrix proteins / 实用口腔医学杂志

Objective:To investigate whether enamel matrix proteins (EMPs) has osteoinductive property. Methods: EMPs and propylene glycol alginate (PGA) in the size of 2 mm3 were placed into calf muscle of 8 SD rats on experiment side. PGA was placed on control side. The rats were sacrificed four weeks after operation. The calf samples were examined by histological observation.Result:Bone-like and/or cartilage-like tissues was not observed in all the operated calf muscles. Conclusion:EMPs are not osteoinductive.