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Purpose: To evaluate, in vitro, the efficiency of a novel apparatus to test the adherence and penetration of bacteria on different membranes for guided regeneration. Methodology: To create the 3D device, Computer Aided Design/Computer Aided Manufacturing (CAD/CAM) systems were used. Three types of biomaterials were tested (n = 6): (DT) a collagen membrane; (DS) a polymer membrane; and (LP) a dense polytetrafluoroethylene barrier. The biomaterials were adapted to the apparatuses and challenged with two different monospecies bacterial culture of A. actinomycetemcomitans b and S. mutans. After 2 h, bacterial adherence and penetration were quantified by counting the number of colony-forming units (CFUs). Two specimens from each group were used for image analysis using Confocal Laser Scanning Microscopy. Statistical analysis was performed. Findings: The DS group had a higher adherence of S. mutans compared to A. actinomycetemcomitans b (p = 0.05). There was less adherence of A. actinomycetemcomitans b in the DS group, compared to the LP (p = 0.011) and DT (p < 0.001) groups. Only the membranes allowed penetration, which was blocked by barriers. The DT group allowed a greater penetration of S. mutans to occur compared to A. actinomycetemcomitans b (p = 0.009), which showed a higher penetration into the DS membranes compared to S. mutans (p = 0.016). The penetration of A. actinomycetemcomitans b through DS was higher compared to its penetration through DT and LP (p < 0.01 for both). DT and DS allowed a greater penetration of S. mutans to occur compared to LP, which prevented both bacterial species from penetrating. Conclusion: The apparatus allowed for the settlement and complete sealing of the biomaterials, enabling standardization.

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The current literature on guided bone regeneration (GBR) and guided tissue regeneration (GTR) membrane contamination reports that the physicochemical characteristics of these biomaterials might influence affinity to bacteria, which appears to be a major drawback for the clinical outcome of the regenerative procedures. Thus, this study aimed to evaluate, in vitro, a multispecies biofilm adherence and passage of bacteria through different types of commercially available membranes for GTR/GBR. Four types of membranes were tested (n=12): LC) Lumina Coat®; JS) Jason®; BG) Biogide®; and LP) Lumina PTFE®. Aluminum foil (AL) simulated an impermeable barrier and was used as the control. The membranes were adapted to specific apparatus and challenged with a mixed bacterial culture composed of A. actinomycetemcomitans b, S. mutans, S. mitis, and A. israelii. After 2 h or 7 days, bacterial adhesion and passage of bacteria were evaluated through CFU counting, which was analyzed by two-way ANOVA e post hoc Tukey, at a 5% significance level. Representative areas of two membranes of each group were analyzed through scanning electron microscopy (SEM) to assess the morphology and organization of the biofilm over the membrane fibers. LC and LP presented similar values of adhered bacterial cells (p > 0.05), significantly inferior when compared to the other groups, in both time points (p < 0.05). All the tested groups were permeable to bacterial cells, with no significant difference between the trial period of 2 h and 7 days (p > 0.05). SEM analyses demonstrated that adhered bacteria number increased throughout the time points (2 h < 7 days). Commercially available biological membranes demonstrated intense bacterial adherence and passage of bacteria, which increased throughout the trial period.

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Abstract The current literature on guided bone regeneration (GBR) and guided tissue regeneration (GTR) membrane contamination reports that the physicochemical characteristics of these biomaterials might influence affinity to bacteria, which appears to be a major drawback for the clinical outcome of the regenerative procedures. Thus, this study aimed to evaluate, in vitro, a multispecies biofilm adherence and passage of bacteria through different types of commercially available membranes for GTR/GBR. Four types of membranes were tested (n=12): LC) Lumina Coat®; JS) Jason®; BG) Biogide®; and LP) Lumina PTFE®. Aluminum foil (AL) simulated an impermeable barrier and was used as the control. The membranes were adapted to specific apparatus and challenged with a mixed bacterial culture composed of A. actinomycetemcomitans b, S. mutans, S. mitis, and A. israelii. After 2 h or 7 days, bacterial adhesion and passage of bacteria were evaluated through CFU counting, which was analyzed by two-way ANOVA e post hoc Tukey, at a 5% significance level. Representative areas of two membranes of each group were analyzed through scanning electron microscopy (SEM) to assess the morphology and organization of the biofilm over the membrane fibers. LC and LP presented similar values of adhered bacterial cells (p > 0.05), significantly inferior when compared to the other groups, in both time points (p < 0.05). All the tested groups were permeable to bacterial cells, with no significant difference between the trial period of 2 h and 7 days (p > 0.05). SEM analyses demonstrated that adhered bacteria number increased throughout the time points (2 h < 7 days). Commercially available biological membranes demonstrated intense bacterial adherence and passage of bacteria, which increased throughout the trial period.

Resumo O objetivo deste estudo foi avaliar, in vitro, a aderência do biofilme multiespécie e a passagem de bactérias através dos diferentes tipos de membranas disponíveis comercialmente para RTG/ROG. Quatro tipos de membranas foram testados (n=12): LC) Lumina Coat®; JS) Jason®; BG) Biogide®; e LP) Lumina PTFE®. Papel alumínio (AL) simulou uma barreira impermeável e foi usado como controle negativo. As membranas foram adaptadas à um aparato específico e desafiadas com uma cultura bacteriana mista composta de A. actinomycetemcomitans b, S. mutans, S. mitis, e A. israelii. Após 2 h ou 7 dias, a aderência e passagem bacteriana foi avaliada através da contagem de UFCs. Duas membranas de cada grupo foram analisadas através da microscopia eletrônica de varredura (MEV). LC e LP apresentaram valores semelhantes de células bacterianas aderidas (p < 0.05), significativamente inferiores quando comparados aos outros grupos, em ambos os períodos experimentais (p < 0.05). Desde a análise inicial, todos os grupos testados foram permeáveis às células bacterianas, sem diferença significativa entre o período experimental de 2 h e 7 dias (p > 0.05). As análises em MEV demonstraram que o número de bactérias aderidas aumentou com o tempo (2 h < 7 days). Membranas biológicas comercialmente disponíveis demonstraram intensa aderência bacteriana e passagem de bactérias, que aumentou durante os períodos experimentais.

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The aim of this integrative review was to identify whether alternative scaffolds used in regenerative endodontics contribute to better root development, in relation to the increase in root length and thickness of dentin walls, compared with blood clot (BC) scaffolds. The literature search was conducted in PubMed, SciELO and Lilacs databases, using descriptors related to the topic. After applying the eligibility criteria, 11 articles were selected and analyzed according to the proposed aim. Five clinical and six in vivo studies, conducted in animals, compared different types of alternative scaffolds with BCs, with emphasis on platelet-rich plasma (PRP) and platelet-rich fibrin (PRF). All scaffolds, alternative or BC, promoted an increase in root length and dentin wall thickness, with varying percentages of increase between studies. In general, there was a significant increase in root length and dentin thickness promoted by PRF and PRP scaffolds, compared with BC. It was concluded that the majority of the scaffolds tested contributed to the increase in root length and thickness of dentin walls, with emphasis on PRF and PRP.

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Abstract The aim of this integrative review was to identify whether alternative scaffolds used in regenerative endodontics contribute to better root development, in relation to the increase in root length and thickness of dentin walls, compared with blood clot (BC) scaffolds. The literature search was conducted in PubMed, SciELO and Lilacs databases, using descriptors related to the topic. After applying the eligibility criteria, 11 articles were selected and analyzed according to the proposed aim. Five clinical and six in vivo studies, conducted in animals, compared different types of alternative scaffolds with BCs, with emphasis on platelet-rich plasma (PRP) and platelet-rich fibrin (PRF). All scaffolds, alternative or BC, promoted an increase in root length and dentin wall thickness, with varying percentages of increase between studies. In general, there was a significant increase in root length and dentin thickness promoted by PRF and PRP scaffolds, compared with BC. It was concluded that the majority of the scaffolds tested contributed to the increase in root length and thickness of dentin walls, with emphasis on PRF and PRP.

Resumo O objetivo desta revisão integrativa foi identificar se os scaffolds alternativos utilizados em endodontia regenerativa contribuem para um melhor desenvolvimento radicular, em relação ao aumento do comprimento e espessura das paredes da dentina, em comparação com os scaffolds de coágulo sanguíneo (BC). A pesquisa bibliográfica foi realizada nas bases de dados PubMed, SciELO e Lilacs, utilizando descritores relacionados ao tema. Após a aplicação dos critérios de elegibilidade, 11 artigos foram selecionados e analisados de acordo com o objetivo proposto. Cinco estudos clínicos e seis in vivo, realizados em animais, compararam diferentes tipos de scaffolds alternativos com BCs, com ênfase no plasma rico em plaquetas (PRP) e fibrina rica em plaquetas (PRF). Todos os scaffolds, alternativos ou BC, promoveram um aumento no comprimento da raiz e na espessura da parede dentinária, com percentuais variáveis de aumento entre os estudos. Em geral, houve um aumento significativo do comprimento da raiz e da espessura da dentina promovido pelos scaffolds PRF e PRP, em comparação com a BC. Concluiu-se que a maioria dos scaffolds testados contribuiu para o aumento do comprimento das raízes e da espessura das paredes dentinárias, com ênfase em PRF e PRP.

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Objetivo: Avaliar e comparar os efeitos antimicrobiano e antibiofilme, e a citotoxicidade promovida pela associação do hidróxido de cálcio ao óleo essencial de Melaleuca alternifolia (MA), em diferentes concentrações, e ao propilenoglicol (PG). Métodos: As seguintes medicações compuseram os grupos experimentais: G1) HC/MA 1%; G2) HC/MA 5%; G3) HC/MA 10%; G4) HC/MA 20%; e G5) HC/PG. Solução salina 0,85% e meio DMEM serviram como controle nos testes antimicrobianos e de citotoxidade em fibroblastos do ligamento periodontal humano (FbLP), respectivamente. A atividade antimicrobiana (n = 12) foi avaliada por meio do teste de difusão em ágar. O efeito antibiofilme (n = 12) imediato das medicações foi avaliado por meio do teste de viabilidade bacteriana em biofilmes de 72 horas de E. faecalis, formados sobre discos de dentina e tratados por sete dias com as medicações. Após a coleta microbiológica do biofilme remanescente, os discos de dentina foram imersos em meio estéril e armazenados por mais sete dias, para a análise do efeito antibiofilme residual das medicações, quando nova coleta microbiológica foi realizada. A atividade metabólica de FbLP foi avaliada por meio do ensaio colorimétrico MTS (n = 9). Os valores médios dos halos de inibição, em mm, das unidades formadoras de colônia, e o percentual de atividade metabólica celular foram analisados pelos testes Kruskal-Wallis e post hoc Dunn (α = 5%). Resultados:Todas as medicações experimentais apresentaram superior ação antimicrobiana e antibiofilme comparadas ao controle, solução salina (p < 0,05), e mantiveram viáveis os FbLP, semelhante ao controle DMEM (p > 0,05). Conclusão: A associação do óleo essencial de Melaleuca alternifolia, nas concentrações de 1%, 5%, 10% e 20%, ao hidróxido de cálcio promoveu excelente ação antimicrobiana, antibiofilme e biocompatibilidade com fibroblastos, de forma semelhante à associação com propilenoglicol.

Aim:To evaluate and compare the antimicrobial and antibiofilm effect, as well as the cytotoxicity of calcium hydroxide (CH) associated with the Melaleuca alternifolia (MA)essential oil, in different concentrations, and with propylene glycol. Methods: The following medications composed the experimental groups: G1) CH/MA 1%; G2) CH/MA 5%; G3) CH/MA 10%; G4) CH/MA 20%; and G5) CH/PG. Saline solution and culture medium DMEM were used as a control in antimicrobial and cytotoxicity tests in human periodontal ligament fibroblasts (PDLF), respectively. The antimicrobial activity (n = 12) was evaluated by the disk-diffusion agar method. The immediate antibiofilm effect (n = 12) of the medications was evaluated for bacterial viability in 72 hours-biofilms of E. faecalis, formed on the dentin disc surface and treated for seven days with medications. After microbiological sampling of the remaining biofilm, the dentin discs were immersed in sterile culture medium and stored for another seven days, for analysis of the residual antibiofilm effect of the medications, when a new microbiological sampling was performed. PDLF viability was evaluated by MTS colorimetric assay (n = 9). The mean values of the inhibition halos, in mm, the colony forming units, and the metabolic cell activity percentage were analyzed by means of Kruskal-Wallis and post hoc Dunn (α = 5%) tests. Results:All of the experimental medications presented higher antimicrobial and antibiofilm effects, when compared to the saline solution control (p < 0.05), and maintained the PDLF feasible, similar to the DMEM control (p > 0.05). Conclusions:The association of the Melaleuca alternifolia essential oil, at concentrations of 1%, 5%, 10%, and 20%, with calcium hydroxide promoted an excellent antimicrobial and antibiofilm activity, and biocompatibility with fibroblasts, similarly to the association with propylene glycol.

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