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).

Self-assembly and mineralization of full-length human amelogenin and its functional fragments / 华西口腔医学杂志

OBJECTIVES@#To investigate the dynamic process of the self-assembly behaviors of a full-length human amelogenin (AM) and its functional fragments tyrosine-rich amelogenin peptide (TRAP) and leucine-rich amelogenin peptide(LRAP) @*METHODS@#The full-length human AM and its functional fragments, TRAP and LRAP, were reassembled and purified @*RESULTS@#When pH=8, the full-length human AM and TRAP assembly started spontaneously and formed "nanospheres" after 15 min.The nanospheres formed by TRAP existed independently, with a uniform size but without obvious internal structures. The full-length AM was assembled hierarchically, which formed "nanospheres" and further extended in all directions, formed a chain structure, and then aggregated into a net. The self-assembly behavior of LRAP was not obvious. Proteins mostly existed in the form of monomers without "nanosphere" formation. Only few oligomers were observed. The full-length AM was induced independently for 3 days to form rod-shaped HA crystals. TRAP and LRAP proteins were added, after 3 days the crystal elongation was obvious in the c axis, but the growth in plane A and plane B was poor.@*CONCLUSIONS@#The self-assembly and mineralization behaviors of full-length human AM, TRAP, and LRAP were consistent with the directional growth mechanism of HA crystals

Expression of enamel proteins in human dental pulp stem cells by the effect of extracellular matrix / Expresión de proteínas de esmalte en células madre de pulpa dental humana por el efecto de matriz extracelular

SUMMARY: Mesenchymal stem cells are present in adult tissues such as the human dental pulp. They are pluripotent and can differentiate into various specialized cell types in vitro through appropriate stimuli. Ameloblasts produce human tooth enamel only during embryonic development before tooth eruption, so endogenous regeneration is not possible. Various efforts have been aimed at generating natural or artificial substitutes for dental enamel with properties similar to the specific components of said tissue. The purpose of this study was to induce human dental pulp stem cells to produce enamel proteins using extracellular matrix derived from the rat tail tendon and pigskin. Primary cultures of human dental pulp stem cells were established and characterized by RT-PCR and immunofluorescence, using mesenchymal cell markers such as CD14, CD40, CD44, CD105, and STRO-1. The cells were then incubated with the extracellular matrix for fourteen days and labeled with specific antibodies to detect the expression of dental enamel proteins such as amelogenin, ameloblastin, enamelisin, tuftelin, and parvalbumin, characteristics of the phenotype of ameloblasts. This work demonstrated a positive effect of the extracellular matrix to induce the expression of enamel proteins in the stem cells of the human dental pulp.

RESUMEN: Las células madre mesenquimales están presentes en los tejidos adultos como la pulpa dental humana. Son pluripotentes y pueden diferenciarse en varios tipos de células especializadas in vitro a través de estímulos adecuados. Los ameloblastos producen esmalte dental humano sólo durante el desarrollo embrionario antes de la erupción dental, por lo que no es posible su regeneración endógena. Varios esfuerzos se han orientado a generar sustitutos naturales o artificiales de esmalte dental con propiedades similares a los componentes específicos de este tejido. El propósito de este estudio fue inducir células madre de pulpa dental humana para producir proteínas del esmalte dental a través del estímulo de matriz extracelular derivada del tendón de la cola de rata y piel de cerdo. Se establecieron cultivos primarios de células madre de pulpa dental humana y se caracterizaron por RT-PCR e inmunofluorescencia utilizando marcadores de células mesenquimales como CD14, CD40, CD44, CD105 y STRO-1. Posteriormente, las células se incubaron con matriz extracelular durante un período de catorce días y se marcaron con anticuerpos específicos para detectar la expresión de proteínas de esmalte dental como amelogenina, ameloblastina, enamelisina, tuftelina y parvalbúmina, las cuales son características del fenotipo de ameloblastos. Este trabajo demostró el efecto positivo que tiene el empleo de la matriz extracelular para inducir la expresión de proteínas de esmalte en las células pluripotenciales de la pulpa dental humana.

The effect of enamel matrix derivatives on root coverage: a 12-month follow-up of a randomized clinical trial

Abstract Subepithelial connective tissue grafts (SCTGs) with a coronally advanced flap (CAF) are accepted as the gold standard for covering denuded root surfaces. In recent years, enamel matrix derivatives (EMDs) have been used for their regenerative potential in periodontics. The aim of this split-mouth and randomized controlled study was to assess the clinical and aesthetical impacts of EMD application in combination with SCTG+CAF in patients with Miller's Class I and II gingival recessions in contralateral canines of the maxilla. Participants who underwent SCTG+CAF+EMD application were identified as the test group (n = 19) and those who underwent SCTG+CAF as control group (n = 19). The outcome parameters were recession depth/width, root coverage percentage, and root coverage aesthetic score (RES). RES was evaluated by two calibrated blind periodontists one year after the treatment. Statistically significant root coverage percentage was observed at one year post-treatment for both groups (p < 0.05). However, significant differences between the groups were not observed in terms of total RES and complete root coverage rate (p > 0.05). The test group had significantly better results than the control according to the soft tissue texture and mucogingival junction alignment results (p < 0.05). These results indicate that EMDs contribute to the healing of soft tissue without scarring. As a result of better wound healing, the EMD-added group exhibited better results in terms of the harmony of the mucogingival junction between adjacent teeth. This paper is the first split-mouth study in which SCTG+CAF and SCTG+CAF+EMD were compared using RES in bilateral canines.

Enamel biomimetics-fiction or future of dentistry / 国际口腔科学杂志·英文版

Tooth enamel is a complex mineralized tissue consisting of long and parallel apatite crystals configured into decussating enamel rods. In recent years, multiple approaches have been introduced to generate or regenerate this highly attractive biomaterial characterized by great mechanical strength paired with relative resilience and tissue compatibility. In the present review, we discuss five pathways toward enamel tissue engineering, (i) enamel synthesis using physico-chemical means, (ii) protein matrix-guided enamel crystal growth, (iii) enamel surface remineralization, (iv) cell-based enamel engineering, and (v) biological enamel regeneration based on de novo induction of tooth morphogenesis. So far, physical synthesis approaches using extreme environmental conditions such as pH, heat and pressure have resulted in the formation of enamel-like crystal assemblies. Biochemical methods relying on enamel proteins as templating matrices have aided the growth of elongated calcium phosphate crystals. To illustrate the validity of this biochemical approach we have successfully grown enamel-like apatite crystals organized into decussating enamel rods using an organic enamel protein matrix. Other studies reviewed here have employed amelogenin-derived peptides or self-assembling dendrimers to re-mineralize mineral-depleted white lesions on tooth surfaces. So far, cell-based enamel tissue engineering has been hampered by the limitations of presently existing ameloblast cell lines. Going forward, these limitations may be overcome by new cell culture technologies. Finally, whole-tooth regeneration through reactivation of the signaling pathways triggered during natural enamel development represents a biological avenue toward faithful enamel regeneration. In the present review we have summarized the state of the art in enamel tissue engineering and provided novel insights into future opportunities to regenerate this arguably most fascinating of all dental tissues.

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®

Expression of enamel proteins in rough endoplasmic reticulum and golgi complex in human dental germs / Expresión de proteínas del esmalte en retículo endoplásmico rugoso y complexo golgiensis en gérmenes dentales humanos

Tooth enamel is the hardest tissue in the body. The organic matrix configuration is provided by the main proteins amelogenin, ameloblastin and enamelysin (MMP20), an enzyme that helps to shape the matrix. The aim of this study was to determine by histochemistry the expression of amelogenin and enamelysin through the rough endoplasmic reticulum in the late stages of amelogenesis, and its expression in the Complexus golgiensis (Golgi complex / Golgi apparatus) in the early stages in human fetuses. In early stages a colocalization of both proteins inside the Golgi apparatus was found, being more evident the relationship between Golgi and amelogenin (99.92 %). In the late stage, a colocalization of both proteins and rugged endoplasmic reticulum was found. With enamelysin being more evident in relation with rough endoplasmic reticulum (99.95 %). Our findings demonstrated the presence of amelogenin and enamelysin in odontoblast and ameloblast. However, the presence of these two proteins in odontoblast remains unknown.

El esmalte dental es el tejido más duro del cuerpo. La configuración de la matriz orgánica es proporcionada por las proteínas principales amelogenina, ameloblastina y enamelisina (MMP20), una enzima que ayuda a dar forma a la matriz. El objetivo de este estudio fue determinar mediante histoquímica la expresión de amelogenina y enamelisina a través del retículo endoplasmático rugoso en las últimas etapas de la amelogénesis , y su expresión en el Complexo golgiensis en las primeras etapas de formación en fetos humanos. En las primeras etapas se observó colocalización de ambas proteínas en el interior del Complexo golgiensis, siendo más evidente la relación entre Golgi y amelogenina (99,92 %). En la última etapa, se identificó una colocalización de ambas proteínas y retículo endoplásmico rugoso. Resulto más evidente la enamelisina en relación con el retículo endoplasmático rugoso (99,95 %). Nuestros resultados demostraron la presencia de amelogenina y enamelisina en odontoblastos y ameloblastos, sin embargo se desconoce la presencia de estas dos proteínas en odontoblastos.

Expresión de tuftelina en gérmenes dentales humanos / Expression of tuftelin in human dental germs

La tuftelina es una proteína secretada en la matriz adamantina en desarrollo durante la formación del esmalte. Su función continúa sin esclarecerse, aunque se presume que juega un papel importante en la biomineralización de esmalte y dentina, así como en el desarrollo del órgano dental. Con el presente estudio se identificó su localización en las diferentes estructuras de gérmenes dentales de fetos humanos, conforme a los resultados se observó su expresión en el estadio pre-secretor observándose en el citoplasma de los ameloblastos, retículo estrellado, papila dental, así como en el estrato intermedio; en el secretor se identificó principalmente en la unión amelodentinaria, y en la superficie externa del esmalte, observando una marcada expresión de la proteína en la porción basal del proceso odontoblástico, pero no en la matriz extracelular de la dentina. De acuerdo a los resultados obtenidos se puede considerar que su expresión se presenta tanto en la amelogénesis, como en la odontogénesis en tejidos sin mineralizar.

The tuftelin is a secreted protein in the adamantine matrix in developing during the enamel formation. Its function continues unclarified, although it plays a role in the biomineralization of the dental organ. With the present studio the location was identified in the different structures of dental germs from human fetuses, according to the results it was observed the expression in the pre-secretor stage being observed in the cytoplasm of ameloblasts, stellate reticulum, dental papilla, also in the intermediate stratum; in the secretor it was mainly identified in the amelodentinal junction and in the outer surface of enamel, observing a marked expression of the protein in the basal portion of the odontoblastic process, but not in the extracellular matrix of the dentine. According to the results obtained it can be considered that its expression occurs in both amelogenesis and odontegenesis in unmineralized tissues.

O impacto clínico e imunológico da matriz derivada de esmalte na cicatrização de cirurgias para recobrimento radicular / Clinical and imunological impact of enamel matriz derivative in root covering surgeries

O objetivo do presente estudo foi avaliar o impacto clínico e imunológico da matriz derivada de esmalte (EMD) na cicatrização periodontal, após a realização de cirurgias de recobrimento radicular. Dezesseis pacientes, com média de idade de 47,7 anos (DP=7,0), foram submetidos à cirurgia de recobrimento radicular com enxerto de tecido conjuntivo subepitelial associado ou não à matriz derivada de esmalte. O estudo seguiu o modelo de boca dividida e randomizado para os lados teste e controle. Os seguintes parâmetros clínicos foram analisados no dia da cirurgia e 6 meses após o procedimento cirúrgico: altura da recessão (AR), largura da faixa de gengiva queratinizada (LFG), largura da recessão (LR), profundidade de sondagem (PS), nível de inserção clínica (NIC) e espessura gengival (EG). Além da análise clínica, foram realizadas avaliações dos seguintes marcadores inflamatórios: IL-1ß, IL -1RA, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL12, IL-13, IL-15, IL-17, EOTAXIN, FGF, G-CSF, GM-CSF, IFN-, IP-10, MCP-1, MIP 1α, PDGF-bb, MIP 1ß, RANTES, TNF-α e VEGF, no fluido gengival no dia da cirurgia e com 7 e 14 dias de pós-operatório. Análise da dor pós-operatória e a análise estética também foram realizadas. Os resultados demonstraram que o lado tratado com EMD resultou em percentual de cobertura radicular (p=0,03) significantemente maior quando comparado com o lado controle. Setenta e cinco por cento dos sítios apresentaram cobertura radicular completa no lado teste e 43,8% no controle, em 6 meses. O EMD não trouxe resultados significantes para aumento de espessura gengival e ganho de faixa de tecido queratinizado. As análises imunológicas mostraram um aumento de VEGF (p=0,03) e PDGF (p=0,03) aos 14 dias de pós-operatório com o EMD e altas expressões de mediadores inflamatórios como IL-1ß (p=0,04; p=0,03), IL-6 (p=0,05; p=0,05), TNF-α (p=0,02; p=0,02), MIP-1α (p=0,02; p=0,05) com 7 dias de pós-operatório, em ambos os grupos, teste e controle, respectivamente. Não houve diferença entre os grupos na avaliação da dor pós-operatória e foi demonstrada uma diferença significante entre a avaliação da estética final do paciente daquela realizada por um profissional calibrado e experiente. Assim sendo, concluímos que o EMD parece ter impacto clínico e imunológico, nas cirurgias de recobrimento de raízes, com percentuais significantemente maiores de cobertura radicular e completa cobertura radicular assim como aumento significantemente maior na liberação de VEGF e PDGF-bb com 14 dias de pós-operatório, sugerindo ser um importante elemento para uma melhor cicatrização.

The aim of the present study was to evaluate the clinical and immunological impact of the enamel-derived matrix (EMD) on periodontal healing, after root-covering surgeries. Sixteen patients, mean age 47.7 years (SD = 7.0), underwent root-graft surgery with graft of subepithelial connective tissue associated or not with enamel-derived matrix. The study followed the split-mouth model and randomized to the sides test and control. The following clinical parameters were analyzed on the day of surgery and 6 months after the surgical procedure: height of the recession (AR), width of keratinized tissue (LFG), width of the recession (LR), probing depth (PS), clinical attachment level (NIC) and gingival thickness (EG). In addition to the clinical analysis, the following inflammatory markers were evaluated: IL-1ß, IL-1RA, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL IL-13, IL-15, IL-17, EOTAXIN, FGF, GCSF, GMCSF, IFN-, IP-10, MCP-1, MIP-1α, PDGF-bb, MIP-1ß, RANTES, TNF-α and VEGF in the gingival crevicular fluid at baseline and at 7 and 14 postoperative days. Postoperative pain analysis and aesthetic analysis were performed. The results showed that the side treated with EMD resulted in a significantly higher percentage of root coverage (p = 0.03) when compared to the control side. Seventy-five percent of the sites had complete root coverage on the test side and 43.8% on the control side in 6 months. The EMD did not bring significant results to increase gingival thickness and gain of width keratinized tissue. Immunological analyzes showed an increase in VEGF (p = 0.03) and PDGF (p = 0.03) at 14 days postoperatively with EMD and high expression of inflammatory mediators such as IL-1ß (p = 0.04) (P = 0.05), MIP-1α (p = 0.05), TNF-α (p = 0.02, p = 0.02) 02, p = 0.05) with 7 postoperative days, in both groups, test and control, respectively. There was no difference between the groups in the evaluation of postoperative pain and a significant difference between the evaluation of the final aesthetic of the patient and the one performed by a calibrated and experienced professional was demonstrated. Thus, we concluded that EMD appears to have a clinical and immunological impact in root coverage surgeries, with significantly greater percentage of root coverage and complete root coverage, as well as a significantly higher increase in the release of VEGF and PDGF-bb with 14 days of postoperative, suggesting to be an important element for a better wound healing.

Study of human leucine-rich amelogenin peptide and its regulation of mineralization by cryogenic transmission electron microscopy / 华西口腔医学杂志

<p><b>OBJECTIVE</b>Recombinant human leucine-rich amelogenin peptide (LRAP) was studied by cryogenic transmission electron microscopy (TEM); evaluation focused on its self-assembly and crystal growth in vitro.</p><p><b>METHODS</b>Human LRAP was recombined through prokaryotic expression vector pCold-SUMO and transformed into Escherichia coli BL21plys to acquire purified proteins. Cryogen TEM recorded assembly and self-assembling of LRAP from pH 3.5 to pH 8.0, and the hydroxyapatite crystal growth in the mixture of LRAP protein solution and artificial saliva was observed using TEM and selected area electron diffraction.</p><p><b>RESULTS</b>More than 90% purity LRAP was expressed, purified and identified as described in methods. LRAP linked into oligomers, nanospheres, nanochains, and microribbons, whereas pH value increased from 3.5 to 8.0. Mature hydroxyapatite crystal growth was guided in artificial saliva filled with calcium phosphate.</p><p><b>CONCLUSIONS</b>LRAP is simplified amelogenin functional domain and conserved the basic characters of amelogenin such as self-assembling and inducing crystallization along c axis. In the area of acellular synthesis of hydroxyapatite using extracellular enamel matrix protein, LRAP is one of candidate repair materials for irregular hard tissue defection.
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Tratamiento de defectos periodontales con matriz derivada del esmalte: serie de casos / Treatment of periodontal defects with enamel matrix derivative: case series

Objetivo: Presentar los resultados clínicos y radiográficos obtenidos a 6 meses del tratamiento de defectos intraóseos con la técnica quirúrgica mínimamente invasiva y el uso de matriz derivada del esmalte sola o en combinación con hueso bovino desmineralizado. Casos clínicos: se seleccionaron 13 sitios con profundidad de sondaje mayor a 5 mm y pérdida ósea vertical. Se evaluaron los siguientes párámetros clínicos: profundidad de sondaje, nivel de inserción clinica y recesión gingival. Los defectos se trataron con matriz derivada del esmalte sola o en combinación con hueso bovino desmineralizado. La profundidad de sondaje inicial promedio era de 6,31 (rango: 5-9) y a los 6 meses, de 3,15 (rango 1-4), lo que significa que la reducción en el promedio de la profundidad de sondaje fue de 3,16. La ganancia en el nivel de inserción clínica fue de 2,9 mm. La evaluación radiográfica evidenció llenado óseo en los sitios tratados. Conclusiones: en la serie de casos presentados, los resultados fueron exitosos y coinciden con los informados en la literatura. La estabilidad del colgajo y el cierre primario son requisitos fundamentales para el éxito y la predictibilidad de cualquier procedimiento regenerativo. Las técnicas quirúrgicas mínimamente invasivas -en comparación con el abordaje tradicional- mejoran notablemente estos parámetros, al reducir los tiempos quirúrgicos y la morbilidad, disminuyendo el malestar del paciente.

Avaliação comparativa entre as técnicas de recobrimento radicular com enxerto de conjuntivo associada ou não à matriz derivada de esmalte (Emdogain) / Comparative evaluation of techniques for root coverage with connective graft associated or not with enamel matrix derivative (Emdogain)

As recessões gengivais são motivos frequentes de visitas aos consultórios odontológicos devido à sua presença causar prejuízos estéticos ao sorriso, além de sensibilidade dolorosa. Existem muitas técnicas descritas na literatura para o recobrimento das raízes expostas, minimizando as consequências das recessões. O objetivo deste trabalho é analisar a técnica de enxerto de tecido conjuntivo e a utilização do Emdogain® para o recobrimento de recessões gengivais e comparar os benefícios de cada uma. Para isso realizou-se uma revisão de literatura a partir de um levantamento bibliográfico utilizando a base de dados da United States National Library of Medicine (PubMed) durante o período entre 2008 e 2013. Foram encontrados 74 estudos no total. Quarenta e sete estudos foram selecionados e coletados. Após aplicação dos critérios de inclusão, vinte e três artigos foram selecionados para a confecção da revisão de literatura e dez passaram por uma análise comparativa a fim de possibilitar a coleta de dados e particularidades de cada estudo. Após a análise comparativa, as duas técnicas demostraram ser efetivas para o tratamento das recessões gengivais e apresentaram taxas semelhantes de cobertura radicular. O uso do enxerto de tecido conjuntivo (ETC) tem a desvantagem de possuir um sítio doador, o que pode causar maior desconforto pós-operatório, por outro lado, as proteínas derivadas da matriz do esmalte (PDME) têm a desvantagem de possuir um alto custo.

Gingival recessions are common reasons for visits to dental offices because of their presence and cause cosmetic damage to smile and soreness. There are many techniques described in the literature for the covering of exposed roots, thus minimizing the effects of recessions. The objective of this work is to analyze the technique of connective tissue graft and use of Emdogain® for coverage of gingival recession and compare the benefits of each. For this we carried out a literature review from a literature survey using the database of the United States National Library of Medicine (PubMed) during the period between 2008 and 2013. Seventy-four studies were found in total. Forty-seven studies were selected and collected. After applying the inclusion criteria, twenty three articles were selected for the preparation of the literature review and ten went through a comparative analysis to enable data collection and particularities of each study. After a comparative analysis, both techniques have shown to be effective for the treatment of gingival recessions and had similar rates of root coverage. The use of the ETC has the disadvantage of having a donor site, which can cause greater discomfort after surgery, on the other hand, PDME has the disadvantage of having a high cost.

In vitro re-hardening of artificial enamel caries lesions using enamel matrix proteins or self-assembling peptides

ABSTRACT Objectives To assess the re-hardening potential of enamel matrix derivatives (EMD) and self-assembling peptides in vitro, hypothesizing that these materials may increase the mineralization of artificial carious lesions and improve hardness profiles. Material and Methods Forty-eight enamel samples were prepared from extracted bovine lower central incisors. After embedding and polishing, nail varnish was applied, leaving a defined test area. One third of this area was covered with a flowable composite (non-demineralized control). The remaining area was demineralized in an acidic buffer solution for 18 d to simulate a carious lesion. Half the demineralized area was then covered with composite (demineralized control), while the last third was left open for three test and one control treatments: (A) Application of enamel-matrix proteins (EMD - lyophilized protein fractions dissolved in acetic acid, Straumann), (B) self-assembling peptides (SAP, Curodont), or (C) amine fluoride solution (Am-F, GABA) for 5 min each. Untreated samples (D) served as control. After treatment, samples were immersed in artificial saliva for four weeks (remineralization phase) and microhardness (Knoop) depth profiles (25-300 µm) were obtained at sections. Two-way ANOVA was calculated to determine differences between the areas (re-hardening or softening). Results Decalcification resulted in significant softening of the subsurface enamel in all groups (A-D). A significant re-hardening up to 125 µm was observed in the EMD and SAP groups. Conclusions This study showed that EMD and SAP were able to improve the hardness profiles when applied to deep demineralized artificial lesions. However, further research is needed to verify and improve this observed effect.

Regeneración tisular guiada en defecto óseo de 3 paredes: enfermedad periodontal y oclusión / Guided tissue regeneration and in a three-walled osseous defect: periodontal disease and occlusion

Relacionar la importancia del éxito en regeneración tisular guiada y el correcto diagnóstico del problema, en este caso enfermedad periodontaly un contacto prematuro en ORC producto de una obturación de amalgama incorrecta. Caso clínico: tratamiento de un defecto infraóseo de3 paredes mediante la utilización de hueso de origen bovino particulado junto con proteínas derivadas de la matriz del esmalte. Tanto los parámetros clínicoscomo los radiográficos fueron evaluados al inicio, en el postquirúrgico inmediato y a los 12 meses. Conclusión: se observó un alto grado de regeneración pasados los 12 meses del tratamiento. Parecería no ser siempre necesaria la utilización de membrana colágena. Las proteínas derivadas de la matrizdel esmalte serían un sustituto de la membrana en algunos casos. Resulta fundamental el chequeo de la situación oclusal en piezas periodontalmente comprometidas...

Avaliação compreensiva da efetividade da proteína da matriz do esmalte no tratamento de recessões gengivais através da revisão sistemática de ensaios pré-clínicos e clínicos / Comprehensive evaluation of the effectiveness of enamel matrix protein in gengival recessions treatment through systematic review of pre-clinical and clinical trials

A proteína da matriz do esmalte (Emdogain, Straumann, Basel, Switzerland) vem ganhando amplo destaque no tratamento das recessões gengivais, por permitir a regeneração dos tecidos de suporte perdidos. O objetivo desta revisão sistemática foi avaliar compreensivamente a efetividade da proteína da matriz do esmalte no recobrimento radicular. Os artigos inclusos nesta revisão foram encontrados através de uma pesquisa eletrônica na base de dados Medline/Pubmed, e a busca estratégica incluiu a seguinte combinação de palavras para estudos pré-clínicos: “periodontal disease AND gingival ressection OR root coverage AND enamel matrix protein AND animal”; e para os estudos clínicos: “periodontal disease AND gingival ressection OR root coverage AND enamel matrix protein”. Não foram inseridos critérios de refinamento de busca. Inicialmente, 64 artigos foram selecionados para análise. Os estudos pré-clínicos incluídos deveriam ser clínicos, pré-clínicos (em animais) e analisar a efetividade da proteína da matriz do esmalte. Os estudos clínicos deveriam apresentar dados numéricos (média e desvio-padrão) inicias e finais relativos às mensurações de nível clínico de inserção (NCI), de profundidade de sondagem (PS), de mucosa queratinizada (MC), de recessão gengival (RG) e período de observação mínimo de 12 meses. Ao final da análise, 18 artigos – cinco estudos pré-clínicos e 13 estudos clínicos longitudinais – foram inclusos e analisados. As evidências científicas analisadas compreensivamente nesta revisão sistemática sugerem que a utilização da proteína da matriz do esmalte é efetiva, apresentando boa estabilidade e previsibilidade no tratamento de recessões gengivais.

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. .