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.

The efficiency of the regeneration of periodontal intrabony defects in East Asians: A systematic review and pooled analysis.

This article aimed to assess the efficacy of periodontal regenerative therapy (PRT) for treating periodontal intrabony defects in East Asians. The systematic review was performed according to the PRISMA guidelines. Literature searches on the PubMed and national medical journal databases, and representative clinical journals of the East Asians were performed on July 31, 2018. Randomized controlled trials, prospective case-control studies, retrospective analyses, and case series receiving regenerative procedures, including barrier membrane (BM) and enamel matrix derivative (EMD) applications with or without bone replacement graft (BRG), with follow-up periods of 6 and 12 months were evaluated. The outcome variables were probing depth (PD) reduction and clinical attachment level (CAL) gain. Twenty studies were included, of which eight were assessed for bias risk. Compared to open flap debridement, PD reduction and CAL gain were superior in all PRTs at both follow-up time points. BM or EMD alone showed equivalent outcomes at 6 months, and CAL gain appeared greater with BM alone at 12 months. BM with BRG showed inferior CAL gain relative to BM alone, but EMD with BRG showed superior CAL gain relative to EMD alone at 12 months. In conclusion, PRT showed improved regenerative outcomes compared with OFD in East Asians, while BM application appeared less efficient than in non-East Asians. BRG supplementation provided additional clinical benefits in EMD application.

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).

Application of enamel matrix derivative in conjunction with non-surgical therapy for treatment of moderate to severe periodontitis: A 12-month, randomized prospective, multicenter study.

BACKGROUND: Treatment of periodontitis aims to halt progressive bone and attachment loss and regenerate periodontal structures. In this study, the effect of using an enamel matrix derivative (EMD) as an adjunct to non-surgical periodontal therapy (test) versus non-surgical therapy alone (control) was evaluated. METHODS: A prospective, split-mouth, multicenter study evaluated scaling and root planing (SRP) with and without EMD in 51 patients presenting with moderate to severe periodontitis (PPD = 5 to 8 mm) in at least 2 pockets per contralateral quadrants within the same arch. The primary outcome variable was change in clinical attachment level (CAL) after 12 months. Secondary variables included probing pocket depth (PPD), bleeding on probing (BoP), gingival margin level, dentin hypersensitivity, and percent of pockets converted to sites no longer requiring surgical treatment. RESULTS: CAL changed significantly (P  < 0.001) from baseline to 12 months for both treatment modalities (test = -2.2 ± 1.5 mm versus control = -2.1 ± 1.3 mm) and similarly for PPD; the difference between groups was not significant. A significant difference, favoring test conditions, was observed in percentage of both healthy PPDs (pockets < 5 mm) and converted pockets (sites no longer requiring surgical treatment); 79.8% of test versus 65.9% of control sites. BoP decreased significantly more (P < 0.05) in test sites (BoP at 17.8% test versus 23.1% control). CONCLUSIONS: Both test and control treatments resulted in significant improvements in CAL and PPD. The adjunct use of EMD with SRP resulted in significantly greater improvements in overall periodontal health with less frequent BoP and a higher number of healthy PPDs.

Platelet-rich plasma, platelet-rich fibrin, and enamel matrix derivative for oral mucosal wound healing.

Different approaches to enhance healing of hard or soft tissues include the use of cytokines and growth factors to modify cellular behaviour. Numerous growth factors are found in autologous blood concentrates - platelet-rich plasma (PRP) and platelet-rich fibrin (PRF). Enamel matrix derivative (EMD) may improve tissue healing via amelogenins. Bilayered collagen matrix (CM) is used for soft tissue augmentation. The aim of the present study was to assess potential benefits of PRP, PRF and EMD in combination with bilayered collagen matrix or CM alone in treatment of oral mucosal defects in rabbits. Twenty-seven New Zealand white rabbits were included in this randomized controlled trial. Artificial oral mucosal defects were treated with one of these five approaches: PRP+CM, PRF+CM, EMD+CM, CM alone, or left untreated as a negative control - CO. The animals were euthanized 1 day, 7 days, or 28 days after surgery and necropsies were harvested. Histological and molecular biological analyses were performed. All defects were healed by day 28. No differences between PRP+CM, PRF+CM, CM alone and CO groups were recorded at any time point. Slower angiogenesis and a higher presence of inflammatory infiltrate were observed in the EMD+CM group 28 days after surgery. Molecular biological analyses did not reveal any statistically significant changes. In conclusion, no improvement in mucosal healing of wounds covered with a collagen membrane and PRP, PRF, or EMD was observed, compared with CM alone or untreated controls.

Periodontal regeneration by leukocyte and platelet-rich fibrin with autogenous bone graft versus enamel matrix derivative with autogenous bone graft in the treatment of periodontal intrabony defects: A randomized non-inferiority trial.

BACKGROUND: Aim of the present study was to ascertain if a combination of leukocyte and platelet-rich fibrin (L-PRF) + autogenous bone graft (ABG) may be a clinically "non-inferior" treatment modality as compared with the association of enamel matrix derivative (EMD) with ABG in the management of intrabony defects (IBDs). METHODS: A total of forty-four patients, exhibiting at least one unfavorable intraosseous defect, were treated by L-PRF associated with ABG (22 patients; test group) or EMD+ABG (control group) in each defect. At baseline and 12 months, a complete clinical and radiographic examination was done. Pre- and post-therapy clinical (probing pocket depth [PPD], clinical attachment level [CAL], gingival recession [GR]) and radiographic (defect Bone level [(DBL)] parameters for the different treatments were compared. To guarantee the test treatment's efficacy 1mm was chosen as non-inferiority margin; for clinical relevance, a second non-inferiority margin = 0.5 mm was set. RESULTS: Clinical and radiographic parameters significantly improved 12 months after surgery in both test and control sites, without inter-groups differences for each measurement. The control group - test group differences for the parameters CAL gain -0.248 mm (-0.618 to 0.122), PPD Reduction -0.397 mm (-0.810 to 0.015), GR Change 0.059 mm (-0.300 to 0.418), DBL Gain -0.250 mm (-0.746 to 0.246) were all within the non-inferiority margin of 0.5 mm. CONCLUSION: Our results suggest that the L-PRF+ABG combined treatment of non-contained IBDs produces non-inferior results in terms of CAL gain, PPD reduction, GR increase and DBL gain in comparison with the EMD+ABG combination.

Does enamel matrix derivative application improve clinical outcomes after semilunar flap surgery? A randomized clinical trial.

OBJECTIVES: To evaluate the treatment of gingival recessions by semilunar coronally positioned flap plus enamel matrix derivative (SCPF + EMD). MATERIALS AND METHODS: Thirty patients with class I localized gingival recession were included. They were randomly allocated in two groups: SCPF + EMD and SCPF. Recession height (RH), recession width (RW), width of keratinized tissue (WKT), thickness of keratinized tissue (TKT), probing depth (PD), and clinical attachment level (CAL) were measured at baseline, 6 and 12 months post-surgery. Patient/professional evaluation of esthetics and root sensitivity was performed. RESULTS: After 12 months, mean root coverage was 1.98 ± 0.33 mm for SCPF + EMD (90.86 ± 14.69%) and 1.85 ± 0.41 mm (79.76 ± 17.44%) for SCPF (p > 0.05). The esthetic evaluation by the patient showed preference for SCPF + EMD. According to the professional evaluation (QCE), the use of EMD decreases the appearance of postoperative scar tissue line. There was a significant reduction in root hypersensitivity with no further complaints by the patients. CONCLUSIONS: The addition of EMD provides significantly better esthetics to SCPF, according to patient and professional assessments. SCPF + EMD is effective but not superior to SCPF for root coverage, after 12 months. CLINICAL RELEVANCE: Previous clinical trials showed that the combination of EMD with coronally advanced flaps may enhance the outcome of root coverage. There is a lack of studies testing the combination of EMD with SCPF. The combination SCPF + EMD provides better esthetics when compared to the SCPF and is effective, but not superior, to SCPF for root coverage, after 12 months. TRIAL REGISTRATION: NCT02459704.

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.

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.

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.

Alternatives to connective tissue graft in the treatment of localized gingival recessions: A systematic review.

AIM: The aim of this Systematic Review (SR) was to assess the clinical efficacy of alternatives procedures; Acellular Dermal Matrix (ADM), Xenogeneic Collagen Matrix (XCM), Enamel Matrix Derivative (EMD) and Platelet Rich Fibrin (PRF), compared to conventional procedures in the treatment of localized gingival recessions. MATERIAL AND METHODS: Electronic searches were performed to identify randomized clinical trials (RCTs) on treatment of single gingival recession with at least 6 months of follow-up. Applying guidelines of the Preferred Reporting Items for Systematic Review and Meta-Analyses statement (PRISMA). The risk of bias was assessed using the Cochrane Collaboration's Risk of Bias tool. RESULTS: Eighteen randomized controlled trials (RCTs) with a total of 390 treated patients (606 recessions) were included. This systematic review showed that: Coronally Advanced Flap (CAF) in conjunction with ADM was significantly better than CAF alone, while the comparison between CAF+ADM and CTG was affected by large uncertainty. The CAF+EMD was significantly better than CAF alone, whereas the comparison between CAF+EMD and CTG was affected by large uncertainty. No significant difference was recorded when comparing CAF+XCM with CAF alone, and the comparison between CAF+XCM and CTG was affected by large uncertainty. The comparison between PRF and others technique was affected by large uncertainty. CONCLUSION: ADM, XCM and EMD assisted to CAF might be considered alternatives of CTG in the treatment of Miller class I and II gingival recession.

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.

Furcation Therapy With Enamel Matrix Derivative: Effects on the Subgingival Microbiome.

BACKGROUND: Although enamel matrix derivative (EMD) has been used to promote periodontal regeneration, little is known of its effect on the microbiome. Therefore, this investigation aims to identify changes in periodontal microbiome after treatment with EMD using a deep-sequencing approach. METHODS: Thirty-nine patients with mandibular Class II buccal furcation defects were randomized to beta-tricalcium-phosphate/hydroxyapatite graft (BONE group), EMD+BONE, or EMD alone. Plaque was collected from furcation defects at baseline and 3 and 6 months post-treatment. Bacterial DNA was analyzed using terminal restriction fragment length polymorphism and 16S pyrotag sequencing, resulting in 169,000 classifiable sequences being compared with the Human Oral Microbiome Database. Statistical comparisons were made using parametric tests. RESULTS: At baseline, a total of 422 species were identified from the 39 defects, belonging to Fusobacterium, Pseudomonas, Streptococcus, Filifactor, and Parvimonas. All three regenerative procedures predictably altered the disease-associated microbiome, with a restitution of health-compatible species. However, EMD and BONE+EMD groups demonstrated more long-term reductions in a higher number of species than the BONE group (P <0.05), especially disease-associated species, e.g., Selenomonas noxia, F. alocis, and Fusobacterium. CONCLUSIONS: EMD treatment predictably alters a dysbiotic subgingival microbiome, decreasing pathogen richness and increasing commensal abundance. Further investigations are needed to investigate how this impacts regenerative outcomes.

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)

Identification of proteins from human permanent erupted enamel.

Proteins from the extracellular matrix of enamel are highly specific and necessary for proper enamel formation. Most proteins are removed from the matrix by enamel proteases before complete mineralization is achieved; however, some residual protein fragments persist in the mineralized matrix of erupted enamel. So far, only amelogenin peptides obtained by traditional bottom-up proteomics have been recovered and identified in human permanent erupted enamel. In this study, we hypothesize that other enamel-specific proteins are also found in human permanent enamel, by analysing human erupted third molars. Pulverized enamel was used to extract proteins, and the protein extract was subjected directly to liquid-chromatography coupled to tandem mass spectrometry (LC-MS/MS) without a previous trypsin-digestion step. Amelogenin and non-amelogenin proteins (ameloblastin and enamelin) were succesfully identified. The sequences of the naturally occurring peptides of these proteins are reported, finding in particular that most of the peptides from the amelogenin X-isoform come from the tyrosine-rich amelogenin peptide (TRAP) and that some were identified in all specimens. In conclusion, our LC-MS/MS method without trypsin digestion increased the coverage of identification of the enamel proteome from a few amelogenin peptides to a higher number of peptides from three enamel-specific proteins.

Proline-Rich Peptide Mimics Effects of Enamel Matrix Derivative on Rat Oral Mucosa Incisional Wound Healing.

BACKGROUND: Proline-rich peptides have been shown to promote periodontal regeneration. However, their effect on soft tissue wound healing has not yet been investigated. The aim of this study is to evaluate the effect of enamel matrix derivative (EMD), tyrosine-rich amelogenin peptide (TRAP), and a synthetic proline-rich peptide (P2) on acute wound healing after a full-thickness flap procedure in an incisional rat model. METHODS: This experimental study has a split-mouth, randomized, placebo-controlled design. Test and control wounds were created on the palatal mucosa of 54 Sprague-Dawley rats. Wounds were histologically processed, and reepithelialization, leukocyte infiltration, and angiogenesis were assessed at days 1, 3, and 7 post-surgery. RESULTS: EMD and P2 significantly promoted early wound closure at day 1 (P <0.001 and P = 0.004, respectively). EMD maintained a significant acceleration of reepithelialization at day 3 (P = 0.004). Wounds treated by EMD and P2 showed increased angiogenesis during the first 3 days of healing (P = 0.03 and 0.001, respectively). Leukocyte infiltration was decreased in EMD-treated wounds at day 1 (P = 0.03), and P2 and TRAP induced a similar effect at days 3 (P = 0.002 and P <0.0001, respectively) and 7 (P = 0.005 and P <0.001). CONCLUSION: EMD and P2 promoted reepithelialization and neovascularization in full-thickness surgical wounds on rat oral mucosa.

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.

Clinical outcomes following regenerative therapy of non-contained intrabony defects using a deproteinized bovine bone mineral combined with either enamel matrix derivative or collagen membrane.

BACKGROUND: The purpose of this study is to compare clinical outcomes in the treatment of deep non-contained intrabony defects (i.e., with ≥70% 1-wall component and a residual 2- to 3-wall component in the most apical part) using deproteinized bovine bone mineral (DBBM) combined with either enamel matrix protein derivative (EMD) or collagen membrane (CM). METHODS: Forty patients with multiple intrabony defects were enrolled. Only one non-contained defect per patient with an intrabony depth ≥3 mm located in the interproximal area of single- and multirooted teeth was randomly assigned to the treatment with either EMD + DBBM (test: n = 20) or CM + DBBM (control: n = 20). At baseline and after 12 months, clinical parameters including probing depth (PD) and clinical attachment level (CAL) were recorded. The primary outcome variable was the change in CAL between baseline and 12 months. RESULTS: At baseline, the intrabony component of the defects amounted to 6.1 ± 1.9 mm for EMD + DBBM and 6.0 ± 1.9 mm for CM + DBBM sites (P = 0.81). The mean CAL gain at sites treated with EMD + DBBM was not statistically significantly different (P = 0.82) compared with CM + DBBM (3.8 ± 1.5 versus 3.7 ± 1.2 mm). No statistically significant difference (P = 0.62) was observed comparing the frequency of CAL gain ≥4 mm between EMD + DBBM (60%) and CM + DBBM (50%) or comparing the frequency of residual PD ≥6 mm between EMD + DBBM (5%) and CM + DBBM (15%) (P = 0.21). CONCLUSION: Within the limitations of the present study, regenerative therapy using either EMD + DBBM or CM + DBBM yielded comparable clinical outcomes in deep non-contained intrabony defects after 12 months.