Use of checkerboard DNA-DNA hybridization to evaluate the internal contamination of dental implants and comparison of bacterial leakage with cast or pre-machined abutments

To evaluate the checkerboard DNA-DNA hybridization method for detection and quantitation of bacteria from the internal parts of dental implants and to compare bacterial leakage from implants connected either to cast or to pre-machined abutments. Nine plastic abutments cast in a Ni-Cr alloy and nine pre-machined Co-Cr alloy abutments with plastic sleeves cast in Ni-Cr were connected to Branemark-compatible implants. A group of nine implants was used as control. The implants were inoculated with 3 mu l of a solution containing 10(8) cells/ml of Streptococcus sobrinus. Bacterial samples were immediately collected from the control implants while assemblies were completely immersed in 5 ml of sterile Tripty Soy Broth (TSB) medium. After 14 days of anaerobic incubation, occurrence of leakage at the implant-abutment interface was evaluated by assessing contamination of the TSB medium. Internal contamination of the implants was evaluated with the checkerboard DNA-DNA hybridization method. DNA-DNA hybridization was sensitive enough to detect and quantify the microorganism from the internal parts of the implants. No differences in leakage and in internal contamination were found between cast and pre-machined abutments. Bacterial scores in the control group were significantly higher than in the other groups (P < 0.05). Bacterial leakage through the implant-abutment interface does not significantly differ when cast or pre-machined abutments are used. The checkerboard DNA-DNA hybridization technique is suitable for the evaluation of the internal contamination of dental implants although further studies are necessary to validate the use of computational methods for the improvement of the test accuracy. To cite this article:do Nascimento C, Barbosa RES, Issa JPM, Watanabe E, Ito IY, Albuquerque Junior RF. Use of checkerboard DNA-DNA hybridization to evaluate the internal contamination of dental implants and comparison of bacterial leakage with cast or pre-machined abutments.Clin. Oral Impl. Res. 20, 2009; 571-577.doi: 10.1111/j.1600-0501.2008.01663.x.

Bacterial leakage along the implant-abutment interface of premachined or cast components

In recent clinical studies, contamination of the inner parts of dental implants through bacterial penetration along the implant components has been observed. The aim of the present in-vitro study was to investigate leakage of Fusobacterium. nucleatum through the interface between implants and premachined or cast abutments. Both premachined (n = 10) and cast (n = 10) implant abutment assemblies were inoculated with 3.0 mu L of microbial inoculum. The assemblies were completely immersed in 5.0 mL of tryptic soy broth culture medium to observe leakage at the implant-abutment interface after 14 days of anaerobic incubation. Bacterial growth in the medium, indicative of microbial leakage, was found only in 1 out of 9 samples (11.1%) in each group. Both premachined and cast abutments connected to external hexagonal implants provide low percentages of bacterial leakage through the interface in in vitro unloaded conditions if the manufacturer`s instructions and casting procedures are properly followed.

Bacterial leakage evaluation of root canals filled with different endodontic sealers

Objective. The objective of this study was to evaluate the sealing ability of AH Plus, Epiphany, Acroseal, Endofill, and Polifil after active lateral condensation technique, by using a bacterial test, during 64 days.Study design. One hundred bovine incisors were cleaned and shaped; then they were filled with the endodontic sealers and adapted into a microcentrifuge tube. The setup root/microcentrifuge tube was added to glass flasks containing Brain Heart Infusion broth. A culture of Enterococcus faecalis was inserted into the upper chamber of each assembly. Daily leakage was evaluated through the broth turbidity.Results. The results were submitted to statistical analysis (Kaplan-Meier method, Kruskal-Wallis and Dunn tests).Conclusions. AH Plus and Endofill had the worst sealing ability when compared with Polifil, which showed the least leakage. Acroseal and Epiphany showed a tendency toward having an intermediate behavior; however, there was no significant difference among Acroseal, Epiphany, and the other sealers. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2009; 108: e56-e60)

Bacterial leakage in root canals filled with conventional and MTA-based sealers

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

CLSM assessment of tubule penetration and bacterial leakage evaluation of two resin-based sealer

Aim: The aims of this study were to assess the penetration of two endodontic sealers (salicylate and epoxy resin-based sealers) into dentinal tubules using CLSM; and to evaluate the bacterial leakage of roots filled with the same sealers associated with gutta-percha. Material and Methods: For sealer penetrability assessment, thirty bovine roots were instrumented and divided into three groups: AHP: EDTA + filling with AH Plus and gutta-percha (n=10), MTAF: EDTA + filling with MTA Fillapex and gutta-percha (n=10), control group: canals were not irrigated with EDTA and were filled with gutta-percha and AH Plus (n=5) or MTA Fillapex (n=5). Rhodamine B was added to the sealers in order to provide adequate fluorescence. The roots were transversely sectioned 3mm from the apex to enable CLSM analysis. Leakage was evaluated for turbidity of the broth in a split chamber model system for 30 days, using Enterococcus faecalis as a microbial marker. Thirty roots were instrumented and divided in four grupos: AHP: filling with AH Plus and gutta-percha (n=10); MTAF: filling with MTA Fillapex and gutta-percha (n=10); positive control: filling with gutta-percha without sealer (n = 5); negative control: sealing with cyanoacrylate to test the seal of the system (n = 5). Results: The medians for dentinal tubule penetration were 6.8% (AHP) and 6.6% (MTAF) (P = 0.82). The average time for bacterial leakage was 8 days in both experimental groups (P = 0.79). Conclusion: MTA Fillapex and AH Plus presented similar behavior regarding dentinal tubule penetration and bacterial leakage.

Abutment coating with diamond-like-carbon films to reduce implant-abutment bacterial leakage

Backgroud: The influence of diamond-like-carbon (DLC) films on bacterial leakage through the interface between abutments and dental implants of external hexagon (EH) and internal hexagon (IH) was evaluated. Film deposition was performed by PECVD (Plasma Enhanced Chemical Vapor Deposition). Sets of implants and abutments (N=180, n=30) were divided according to the connection design and the treatment of the abutment base: (1) no treatment (control); (2) DLC film deposition, and (3) Ag-DLC film deposition. Under sterile conditions, 1 µL of Enterococcus faecalis was inoculated inside the implants, and abutments were tightened. The sets were tested for immediate external contamination, suspended in test tubes containing sterile culture broth, and followed-up for five days. Turbidity of the broth indicated bacterial leakage. At the end of the period, the abutments were removed and the internal content of the implants was collected with paper points and plated in Petri dishes. They were incubated for 24 h for bacterial viability assessment and colony-forming unit (CFU) counting. Bacterial leakage was analyzed by Chi-square and Fisher exact tests (α=5%). The percentage of bacterial leakage was 16.09% for EH implants and 80.71% for IH implants (P<0.0001). The bacterial load was higher inside these implants (P=0.000). The type of implant significantly influenced the results (P=0.000), whereas the films did not (P=0.487). We concluded that: (1) IH implants showed a higher frequency of bacterial leakage and (2) the DLC and Ag-DLC films did not significantly reduce the frequency of bacterial leakage and bacteria load inside the implants.

Bacterial leakage along the implant-abutment interface: culture and DNA Checkerboard hybridization analyses

Objective Bacterial species have been found harboring the internal surface of dental implants as consequence of their failed connections. The aim of the present study was to compare the detection frequency of bacterial leakage from human saliva through the implantabutment interface, under non-loading conditions, using either DNA Checkerboard or culture method. Materials and methods Thirty dental implants with hexagonal platforms were connected to pre-machined abutments according to the manufacturers specifications. The assemblies were individually incubated in human saliva under anaerobic conditions for 7 similar to days at 37 degrees C. Afterward, contents from the inner parts of the implants were collected and evaluated with either DNA Checkerboard (s similar to=similar to 15) or culture (n similar to=similar to 15). Subsequently, identification and quantitation of bacterial species from saliva and implants were carried out for the group evaluated with the DNA Checkerboard method. Results Both DNA Checkerboard and culture showed positive signals of bacterial leakage in 6 of the 15 evaluated samples. Capnocytophaga gingivalis and Streptococcus mutans were the most frequently detected species harboring the internal surface of the implants followed by Veillonella parvula. Conclusion Occurrence of bacterial leakage along the implantabutment interface is comparably detected with both DNA Checkerboard hybridization and conventional culture methods.

Bacterial leakage through temporary fillings in core buildup composite material - an in vitro study

To evaluate the ability of the provisional filling material Cavit-W alone or in combination with different restorative materials to prevent bacterial leakage through simulated access cavities in a resin buildup material.

Microbial leakage of MTA, Portland cement, Sealapex and zinc oxide-eugenol as root-end filling materials

Objective: The aim of this study was to compare the microbial leakage of mineral trioxide aggregate (MTA), Portland cement (PC), Sealapex and zinc oxide-eugenol (ZOE) as root-end filling materials.Study design: An in vitro microbial leakage test (MLT) with a split chamber was used in this study. A mixture of facultative bacteria and one yeast (S. aureus + E. faecalis + P. aeruginosa + B. subtilis + C. albicans) was placed in the upper chamber and it could only reach the lower chamber containing Brain Heart Infusion broth by way of leakage through the root-end filling. Microbial leakage was observed daily for 60 days. Sixty maxillary anterior human teeth were randomly assigned to different groups - MTA and PC (gray and white), Sealapex + zinc oxide and ZOE, control groups and subgroups to evaluate the influence of EDTA for smear layer removal. These materials were further evaluated by an agar diffusion test (ADT) to verify their antimicrobial efficacy. Data were analyzed statistically by Kruskal-Wallis and Mann-Whitney test.Results: In the MLT, Sealapex + zinc oxide and ZOE did not show evidence of microbial leakage over the 60-day experimental period. The other materials showed leakage from the 15th day. The presence of smear layer influenced microbial leakage. Microbial inhibition zones were not observed in all samples tested by ADT.Conclusion: Sealapex + zinc oxide and ZOE did not show microbial leakage over the experimental period, whereas it was verified within 15 to 45 days in MTA and Portland cement.

Leakage of Saliva Through the Implant-Abutment Interface: In Vitro Evaluation of Three Different Implant Connections Under Unloaded and Loaded Conditions

Purpose: Bacterial leakage along the implant-abutment interface, with consequent species harboring the inner parts of two-part dental implant systems, has been reported in the literature. The aim of this in vitro study was to evaluate bacterial leakage from human saliva to the internal part of the implants along the implant-abutment interface under loaded and unloaded conditions using DNA Checkerboard. Materials and Methods: Sixty denial implants-20 each of external-hexagon, internal-hexagon, and Morse cone-connection designs-and their conical abutments were used in this study. Each group was subdivided into two groups of 10 loaded and 10 unloaded implants. The assemblies were immersed in human saliva and either (1) loaded with 500,000 cycles at 120 N (experimental group) or (2) incubated in static conditions for 7 days at 35 degrees C (unloaded control group). Results: Microorganisms were found in the internal surfaces of all types of connections. The Morse cone connection presented the lowest count of microorganisms in both the unloaded and loaded groups. Loaded implants presented with higher counts of microorganisms than unloaded implants for external- and internal-hex connections. Conclusion: Bacterial species from human saliva may penetrate along the implant-abutment interface under both unloaded and loaded conditions for all connections evaluated. Morse cone-connection implants showed the lowest counts of microorganisms for both conditions. External- and internal-hex implants showed a higher incidence of bacteria and higher bacterial counts after simulated loading. INT J ORAL MAXILLOFAC IMPLANTS 2012;27:551-560.

Back-scattered electron imaging for leakage analysis of four retrofilling materials

This study aimed to evaluate, ex vivo, the nanoleakage in dentinal tubules, the linear infiltration of silver nitrate in the dentin wall/root-end filling material interface, and the presence of gaps in this interface in root-end cavities filled with 4 filling materials. Forty-eight disto-buccal root canals of maxillary molars were instrumented and filled. Retrograde cavities were prepared with ultrasonic points (apical 2 mm). The samples were divided into 2 control groups (n = 4) and 4 experimental groups (n = 10): Group I white mineral trioxide aggregate (MTA); Group II Super EBA; Group III Portland cement; and Group IV Sealer 26. After 1 week, the specimens were subjected to silver nitrate and prepared for SEM (backscattered electrons). In the apical-apical segment, an area with significantly higher leakage was observed for Super EBA, followed by Portland cement, MTA, and Sealer 26 (P = 0.0054). In the medium and cervical segments, all materials showed the same leakage behavior (P = 0.1815 and P = 0.1723, respectively). The linear infiltration at the dentin wall/root-end filling material interface was higher with Super EBA than the other groups. No differences in the percentage of gaps along the 3 mm of dentin wall/root-end filling material interface between the 4 materials were evident (P > 0.05). Nanoleakage occurred mainly in the apical segment of the samples, and Super EBA showed the highest values. The area and linear leakage were lower in the middle and coronal segments, regardless of the root-end filling material. No material perfectly sealed the root-end cavities, which allowed for the leakage occurrence. Microsc. Res. Tech. 75:796800, 2012. (C) 2011 Wiley Periodicals, Inc.

Infiltração microbiana por Enterococcus faecalis em canais retrobturados com MTA, iRoot SP e Endo CPM Sealer

O objetivo deste trabalho consistiu na análise da infiltração apical em dentes retrobturados por três materiais: MTA, iROOT SP e Endo CPM Sealer. Para tal, foram utilizados 51 dentes humanos extraídos, incisivos centrais superiores, que foram instrumentados manualmente com limas tipo K, pela técnica Crown-down, obturados com compactação lateral e, após serem apicectomizados a 3mm aquém do ápice foram submetidos à retrobturação, com os três materiais propostos. As amostras foram divididas, randomicamente, em três grupos: GI MTA, GII iROOT SP e GIII Endo CPM Sealer, cada grupo com 15 amostras. Os dentes foram inseridos em tubos de eppendorfs, e feitos a impermeabilização do remanescente radicular utilizando duas camadas de cianocrilato, epóxi, e outra camada de esmalte. Em cada eppendorf foi adicionado caldo TSB estéril e uma suspensão de Enterococcos faecalis e adaptado ao frasco de vidro com meio de cultura enterococcosel. A infiltração bacteriana foi verificada pela turvação do meio de cultura. Após a análise no período de 60 dias, podemos concluir que durante esse tempo ocorreram infiltrações no Grupo I, 43,75 % das amostras apresentaram turvamento do meio de cultura demonstrando persistência da infecção. Já no Grupo II, 31,25 % das amostras tiveram crescimento bacteriano. Por fim no Grupo III, 25,00 % houve a infiltração. Grupos controle positivo e negativo para crescimento bacteriano foram realizados (n=3, cada). Os cimentos testados comportaram-se de maneira semelhante frente à infiltração bacteriana durante o período testado.

Complicações microbiológicas na interface implante-pilar

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Medicina Dentária

Implant-abutment gap versus microbial colonization: Clinical significance based on a literature review

Microorganisms from the oral cavity may settle at the implant-abutment interface (IAI). As a result, tissue inflammation could occur around these structures. The databases MEDLINE/PubMed and PubMed Central were used to identify articles published from 1981 through 2012 related to the microbial colonization in the implant-abutment gap and its consequence in terms of crest bone loss and osseointegration. The following considerations could be put forward, with respect to the clinical importance of IAI: (a) the space present at the IAI seems to allow bacterial leakage to occur, in spite of the size of this space; (b) bacterial leakage seems to occur at the IAI, irrespective of the type of connection. More studies are necessary to clarify the relationship between leakage at IAI and abutment connection designs; (c) losses at the peri-implant bone crests cannot be related to the IAI size, since few studies have shown no relationship. Also, the microbial leakage at the IAI cannot be related to the bone crest loss, since there are no articles reporting this relationship; remains controversial the influence of the IAI position on the bone crest losses. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 101B: 1321-1328, 2013. Copyright © 2013 Wiley Periodicals, Inc.

Avaliação da infiltração bacteriana corono-apical de canais radiculares obturados com diferentes tipos de cimentos endodônticos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)