Bacterial translocation and microgap formation at a novel conical indexed implant abutment system for single crowns.

OBJECTIVES: A conometric concept was recently introduced in which conical implant abutments hold the matching crown copings by friction alone, eliminating the need for cement or screws. The aim of this in vitro study was to assess the presence of microgap formation and bacterial leakage at the Acuris conometric restorative interface of three different implant abutment systems. MATERIAL AND METHODS: A total of 75 Acuris samples of three implant-abutment systems (Ankylos, Astra Tech EV, Xive) were subjected to microbiological (n = 60) and scanning electron microscopic (SEM) investigation (n = 15). Bacterial migration into and out of the conical coupling system were analyzed in an anaerobic workstation for 48, 96, 144, and 192 h. Bacterial DNA quantification using qrt-PCR was performed at each time point. The precision of the conometric coupling and internal fit of cemented CAD/CAM crowns on corresponding Acuris TiN copings were determined by means of SEM. RESULTS: qrt-PCR results failed to demonstrate microbial leakage from or into the Acuris system. SEM analysis revealed minute punctate microgaps at the apical aspect of the conometric junction (2.04 to 2.64 µm), while mean cement gaps of 12 to 145 µm were observed at the crown-coping interface. CONCLUSIONS: The prosthetic morse taper connection of all systems examined does not allow bacterial passage. Marginal integrity and internal luting gap between the ceramic crown and the coping remained within the clinically acceptable limits. CLINICAL RELEVANCE: Conometrically seated single crowns provide sufficient sealing efficiency, relocating potential misfits from the crown-abutment interface to the crown-coping interface.

Marginal Adaptation and Microbial Leakage at Conometric Prosthetic Connections for Implant-Supported Single Crowns: An In Vitro Investigation.

Encouraging clinical results were reported on a novel cone-in-cone coupling for the fixation of dental implant-supported crowns (Acuris, Dentsply Sirona Implants, Mölndal, Sweden). However, the presence or absence of a microgap and a potential bacterial leakage at the conometric joint has not yet been investigated. A misfit and a resulting gap between the conometric components could potentially serve as a bacterial reservoir that promotes plaque formation, which in turn may lead to inflammation of the peri-implant tissues. Thus, a two-fold study set-up was designed in order to evaluate the bidirectional translocation of bacteria along conometrically seated single crowns. On conometric abutments filled with a culture suspension of anaerobic bacteria, the corresponding titanium nitride-coated (TiN) caps were fixed by friction. Each system was sterilized and immersed in culture medium to provide an optimal environment for microbial growth. Positive and negative controls were prepared. Specimens were stored in an anaerobic workstation, and total and viable bacterial counts were determined. Every 48 h, samples were taken from the reaction tubes to inoculate blood agar plates and to isolate bacterial DNA for quantification using qrt-PCR. In addition, one Acuris test system was subjected to scanning electron microscopy (SEM) to evaluate the precision of fit of the conometric coupling and marginal crown opening. Throughout the observational period of one week, blood agar plates of the specimens showed no viable bacterial growth. qrt-PCR, likewise, yielded a result approaching zero with an amount of about 0.53 × 10-4 µg/mL DNA. While the luting gap/marginal opening between the TiN-cap and the ceramic crown was within the clinically acceptable range, the SEM analysis failed to identify a measurable microgap at the cone-in-cone junction. Within the limits of the in-vitro study it can be concluded that the Acuris conometric interface does not allow for bacterial translocation under non-dynamic loading conditions.

Influence of margin location and luting material on the amount of undetected cement excess on CAD/CAM implant abutments and cement-retained zirconia crowns: an in-vitro study.

BACKGROUND: The flexibility in designing the submucosal part of CAD/CAM customized implant abutments and the individual positioning of its shoulder line has been suggested to reduce the risk of leaving undetected cement residues, thus preventing adverse effects on peri-implant tissues. A high correlation between excess cement left in the soft tissues and the occurrence of increased biofilm accumulation with sulcular bleeding and/ or suppuration has been reported. This in turn may cause peri-implant inflammation and peri-implant marginal bone loss. The aim of this study was to assess the frequency of cement remnants after the luting of zirconia crowns on CAD/CAM custom molar abutments with different margin levels and to evaluate the impact of the luting material. MATERIAL AND METHODS: A total of 20 titanium molar CAD/CAM implant abutments (BEGO Medical GmbH) with internal taper connection/ internal hex anti-rotation protection, and a convex emergence profile with different margin positions (0, 1, 2 and 3 mm below the mucosa), were virtually designed (Implant Studio, 3Shape) and manufactured. A master cast was scanned, duplicated by a 3D printer and individual gingival masks were produced to simulate peri-implant soft tissues. 20 corresponding zirconia crowns were designed (Cerec 3D, Dentsply Sirona), produced and cemented to the abutments with two different luting materials; a zinc oxide non-eugenol cement (Temp Bond NE) or a methacrylate cement (Panavia V5). To ensure retrievability of the crown/abutment connection, occlusal openings providing access to the abutment screws were designed. Excess cement was thoroughly removed and the crown/abutment units were unscrewed to evaluate the occurrence of cement residues. All the quadrants of each specimen were evaluated for calculation of the ratio between the cement remnant area and the total specimen area using Adobe Photoshop. Spearman analysis was performed to detect correlations between different variables. A two-sided t-test, ANOVA, Mann-Whitney, and Kruskal-Wallis tests were applied to detect differences between the groups. RESULTS: Cement remnants were found in every depth of the crown abutment complex and in almost every area investigated. The amount of cement residues increased as the crown-abutment margin was located more submucosally. Lingual areas were more prone to cement remnants than other surface areas (p = 0.0291). Excess cement was not only found at the margins of the crown-abutment complex, but also underneath (basal) the abutment itself, where cleaning was impossible. No statistical difference in the effect of zinc oxide non-eugenol- and methacrylate cement on the frequency of excess material at the lateral abutment surfaces could be demonstrated in vitro. The proportion of basal abutment aspects covered with cement residues was, however, significantly smaller in Panavia V5 samples with an average of 4.9 ± 3.7% compared to Temp Bond samples with an average of 8.6 ± 5.5%. CONCLUSIONS: Given the results obtained in the present investigation the margin of CAD/CAM molar abutments should be located as coronally as possible to minimize the amount of cement remnants. If an epigingival or supragingival margin location is not feasible due to esthetic concerns, it cannot be recommended to place the margin of molar CAD/CAM abutments deeper than 1.5 mm in the proximal and oral regions.

Pharmacokinetics of Intravenous Finafloxacin in Healthy Volunteers.

Finafloxacin is a novel fluoroquinolone exhibiting enhanced activity under acidic conditions and a broad-spectrum antibacterial profile. The present study assessed the pharmacokinetic properties and the safety and tolerability of finafloxacin following intravenous infusions. In this mixed-parallel-group, crossover study, healthy male and female volunteers received single ascending doses (18 volunteers, 200 to 1,000 mg) or multiple ascending doses (40 volunteers, 600 to 1,000 mg) of finafloxacin or placebo. Plasma and urine samples were collected by a dense sampling scheme to determine the pharmacokinetics of finafloxacin using a noncompartmental approach. Standard safety and tolerability data were documented. Finafloxacin had a volume of distribution of 90 to 127 liters (range) at steady state and 446 to 550 liters at pseudoequilibrium, indicating the elimination of a large fraction before pseudoequilibrium was reached. Areas under the concentration-time curves and maximum plasma concentrations (geometric means) increased slightly more than proportionally (6.73 to 45.9 µg · h/ml and 2.56 to 20.2 µg/ml, respectively), the terminal elimination half-life increased (10.6 to 17.1 h), and the urinary recovery decreased (44.2% to 31.7%) with increasing finafloxacin doses (single doses of 200 to 1,000 mg). The pharmacokinetic profiles suggested multiphasic elimination by both glomerular filtration and saturable tubular secretion. The values of the parameters were similar for single and multiple administrations. The coefficient of variation for the between-subject variability of exposure ranged from 10% (≤600 mg) to 38% (>600 mg). Adverse events were mild and nonspecific, with no dependence of adverse events on dose or treatment (including placebo) being detected. Despite a relatively high interindividual variability at higher doses, the level of exposure following intravenous administration of finafloxacin appears to be predictable. Individual elimination processes should be evaluated in more detail. Finafloxacin exhibited a favorable safety and tolerability profile. (This study has been registered at ClinicalTrials.gov under registration no. NCT01910883.).

Effect of cement type, luting protocol, and ceramic abutment material on the shade of cemented titanium-based lithium disilicate crowns and surrounding peri-implant soft tissue: a spectrophotometric analysis.

PURPOSE: The objective of the study was to analyze the impact of cement, bonding pretreatment, and ceramic abutment material on the overall color results of CAD-CAM ceramic crowns bonded to titanium-based hybrid abutments. MATERIALS AND METHODS: For single implant restoration of a maxillary lateral incisor a total of 51 CAD-CAM-fabricated monolithic lithium disilicate crowns were fabricated and subsequently bonded onto 24 lithium disilicate Ti-base abutments, 24 zirconia Ti-base abutments and 3 resin abutment replicas as a control group. The 48 copings were cemented with three definitive and one provisional cement on both grit-blasted and non-blasted Ti-bases. The color of each restoration and surrounding artificial gingiva was measured spectrophotometrically at predefined measuring points and the CIELAB (ΔEab) color scale values were recorded. RESULTS: The color outcome of ceramic crowns bonded to hybrid abutments and soft tissues was affected differently by cements of different brands. Grit-blasting of Ti-bases prior to cementing CAD-CAM copings affected the color results of all-ceramic crowns. There was a significant difference (P = .038) for the median ΔE value between blasted and non-blasted reconstructions at the cervical aspect of the crown. Full-ceramic crowns on zirconia Ti-base abutments exhibited significantly lower ΔE values below the threshold of visibility (ΔE 1.8). In all subcategories tested, the use of a highly opaque temporary cement demonstrated the lowest median ΔE for both the crown and the artificial gingiva. CONCLUSION: Various cements, core ceramic materials and airborne particle abrasion prior to bonding can adversely affect the color of Ti-base supported ceramic crowns and peri-implant soft tissue. However, zirconia CAD-CAM copings and an opaque cement can effectively mask this darkening.

Retrospective, Multi-center, Cohort Study Evaluating Clinical, Technical, Biological, and Esthetic Outcomes of Implant Supported Zirconia Single Crowns Cemented on Titanium-base Abutments: 7-year Mean Follow-up.

PURPOSE: To retrospectively evaluate the long-term clinical, technical, biological, and esthetic outcomes of implant supported single zirconia crowns (ISCs) intraorally cemented to Ti-base hybrid-abutments up to 16 years after placement. MATERIALS AND METHODS: A total of 63 ISCs (Xive S, Camlog Screw Line, Replace Select TC NP, Branemark MK II, and 3i Osseotite) were evaluated in 36 patients at two different centers. Original Ti-bases were selected and zirconia meso structures and zirconia crowns were designed using CAD/CAM software and then milled from partially stabilized zirconia blocks. After the meso structures were cemented extraorally onto the Ti-bases, the ceramic crowns were intraorally luted to the hybrid abutments. The Ti-base ISC restorations were followed up for up to 16 years, and their clinical, biological, and esthetic outcomes were recorded at distinct time points (T1; T2) at three-year intervals. RESULTS: 36 patients (18 men, 18 women) received 32 ISCs in the anterior region and 31 in the posterior region of the maxilla and mandible. The mean follow-up of the Ti-base ISCs was 6.93 ± 2.60 years. The mean follow-up of the implants amounted to 8.11 ± 3.26 years. No implants were lost during follow-up, resulting in a cumulative implant survival rate of 100%. Abutment screw loosening was observed in two ISCs after one year in service. The overall cumulative restorative survival rate of the Ti-base restorations reached thus 96.83%. At T2 follow-up 24% of the ISCs exhibited an increase in PD despite maintaining clinically healthy peri-implant tissue. An 11% increase in BOP and a 3.17% decrease in PI were recorded. Despite spectrophotometrically measured ΔE values indicating visible discoloration of some restorations and their peri-implant soft tissue, a low incidence of esthetic complications was observed with an average PES/WES score of ≥ 12. No correlation was found between PES (R = -0.25; p = 0.27) and WES (R = -0.18; p = 0.43) scores and digital shade determination. CONCLUSIONS: The results of the present retrospective, multicenter, cohort study indicate satisfactory clinical outcomes for intraorally cemented single zirconia crowns (ISCs) supported by Ti-base hybrid abutments. An overall esthetic superiority of Ti-base ISCs could not be confirmed.

Reconfigurable wavefront sensor for ultrashort pulses.

A highly flexible Shack-Hartmann wavefront sensor for ultrashort pulse diagnostics is presented. The temporal system performance is studied in detail. Reflective operation is enabled by programming tilt-tolerant microaxicons into a liquid-crystal-on-silicon spatial light modulator. Nearly undistorted pulse transfer is obtained by generating nondiffracting needle beams as subbeams. Reproducible wavefront analysis and spatially resolved second-order autocorrelation are demonstrated at incident angles up to 50° and pulse durations down to 6 fs.

Microbiological cleaning and disinfection efficacy of a three-stage ultrasonic processing protocol for CAD-CAM implant abutments.

PURPOSE: Computer-aided design and manufacturing (CAD-CAM) of implant abutments has been shown to result in surface contamination from site-specific milling and fabrication processes. If not removed, these contaminants can have a potentially adverse effect and may trigger inflammatory responses of the peri-implant tissues. The aim of the present study was to evaluate the bacterial disinfection and cleaning efficacy of ultrasonic reprocessing in approved disinfectants to reduce the microbial load of CAD-CAM abutments. MATERIALS AND METHODS: Four different types of custom implant abutments (total N = 32) with eight specimens in each test group (type I to IV) were CAD-CAM manufactured. In two separate contamination experiments, specimens were contaminated with heparinized sheep blood alone and with heparinized sheep blood and the test bacterium Enterococcus faecium. Abutments in the test group were processed according to a three-stage ultrasonic protocol and assessed qualitatively and quantitatively by determination of residual protein. Ultrasonicated specimens contaminated with sheep blood and E. faecium were additionally eluted and the dilutions were incubated on agar plates for seven days. The determined bacterial counts were expressed as colony-forming units (CFU). RESULTS: Ultrasonic reprocessing resulted in a substantial decrease in residual bacterial protein to less than 80 µg and a reduction in microbiota of more than 7 log levels of CFU for all abutment types, exceeding the effect required for disinfection. CONCLUSION: A three-stage ultrasonic cleaning and disinfection protocol results in effective bacterial decontamination. The procedure is reproducible and complies with the standardized reprocessing and disinfection specifications for one- or two-piece CAD-CAM implant abutments.

Optical Efficacy of Titanium Nitride-Coated Abutment Material on Soft Tissue Discoloration: A Spectrophotometric In Vitro Analysis.

PURPOSE: The objective of this investigation was to assess the extent of mucosal discoloration caused by different CAD/CAM abutment materials and to determine the influence of mucosa thickness on the subsequent color, with a particular focus on titanium nitride. MATERIALS AND METHODS: In a pig maxilla, a trapdoor-shaped mucosa flap was prepared unilaterally. Several CAD/CAM abutment materials were used to assess a number of clinical scenarios. Varying mucosa thicknesses were simulated by connective tissue grafts harvested at the contralateral side of the palate, resulting in layer thicknesses of 1.5, 2, and 3 mm. Titanium (Ti), zirconia (ZrO2), and titanium nitride (TiN) served as test specimens with and without ceramic veneering. Color differences (ΔE) and deviations in brightness (L), chroma (C), and hue (H) were determined spectrophotometrically, comparing the measured value of the native tissue and the results obtained with different materials at varying mucosa thicknesses. RESULTS: All tested specimens caused a mucosa discoloration in comparison to the native tissue, diminishing with increasing mucosa thickness. The use of TiN demonstrated the least mucosa discoloration in thin soft tissue of 1.5 mm, with a mean ΔE value of 1.93 (P = .004). While ZrO2 revealed a comparable ΔE value of 2.13 (P = .022) at a tissue thickness of 1.5 mm, Ti showed the highest mucosa discoloration above the visibility threshold of ΔE = 3.1, with a mean ΔE value of 4.07 (P = .002). Ceramic veneering of the Ti samples led to a considerable reduction in soft tissue discoloration, with a resulting ΔE value of 2.2. The veneering of TiN and ZrO2 samples with porcelain, on the other hand, had no noticeable effect on the mucosa color. CONCLUSION: CAD/CAM abutment materials cause an adverse soft tissue color shift that decreases with increasing mucosa thickness. In thin peri-implant mucosa, titanium nitride and zirconia lead to the least discoloration. Due to their positive optical properties and mechanical superiority compared with ceramic abutments, gold-hue titanium nitride-coated CAD/CAM abutments could be a clinical alternative in cases of thin peri-implant mucosa.

Qualitative and Semi-Quantitative Assessment of Processing-Related Surface Contamination of One- and Two-Piece CAD/CAM Abutments before and after Ultrasonic Cleaning.

Manufacturing processes of custom implant abutments may contaminate their surfaces with micro wear deposits and generic pollutants. Such particulate debris, if not removed, might be detrimental and provoke inflammatory reactions in peri-implant tissues. Although regulatory guidelines for adequate cleaning, disinfection, or sterilization exist, there does not appear to be a consistent application and data on the amount and extent of such contaminants is lacking. The aim of the present in vitro study was to evaluate the quality and quantity of processing-related surface contamination of computer-aided design/computer-aided manufacturing (CAD/CAM) abutments in the state of delivery and after ultrasonic cleaning. A total of 28 CAD/CAM monotype and hybrid abutments were cleaned and disinfected applying a three-stage ultrasonic protocol (Finevo protocol). Before and after cleaning, the chemical composition and the contamination of the abutments were assessed using scanning electron microscopy (SEM), dispersive X-ray spectroscopy (EDX), and computer-aided planimetric measurement (CAPM). In the delivery condition, monotype abutments showed a significantly higher amount of debris compared to hybrid abutments (4.86 ± 6.10% vs. 0.03 ± 0.03%, p < 0.001). The polishing process applied in the laboratory after bonding the hybrid abutment components reduces the surface roughness and thus contributes substantially to their purity. The extent of contamination caused by computer-aided manufacturing of custom abutments can be substantially minimized using a three-stage ultrasonic protocol.

The Influence of an Ultrasonic Cleaning Protocol for CAD/CAM Abutment Surfaces on Cell Viability and Inflammatory Response In Vitro.

BACKGROUND/AIM: To evaluate the effect of an ultrasonic cleaning and disinfection method for CAD/CAM abutment surfaces on cell viability and inflammatory response in vitro. MATERIALS AND METHODS: Untreated and manually polished surfaces of CAD/CAM generated titanium and zirconia disks were randomly assigned, either to a 3-step ultrasonic cleaning and disinfection process (test: TiUF, TiPF, ZrUF, ZrPF) or to 30 sec steam cleaning (control: TiUS, TiPS, ZrUS, ZrPS). Pre-cleaning surface analyses using scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), and surface profilometry were performed. Human gingival fibroblasts (HGFs) were cultured on test and control specimens and subsequently examined for cell viability and inflammatory response. Expression of acute inflammatory cytokine interleukin (IL)-6 and vascular endothelial growth factor A (VEGFA) were assessed by means of RT-qPCR. RESULTS: Cells on all specimens exhibited a satisfactory viability, indicating firm attachment. Cells on polished zirconia samples, cleaned by means of sonication (ZrPF), exhibited significantly higher viability than cells on the same material cleaned by steam (ZrPS), p=0.019. For all other three material/ surface treatment combinations (TiU, TiP, ZrU), no such difference was observed between the cleaning methods. The messenger ribonucleic acid (mRNA) levels of IL-6 and VEGFA were between 50 and 105% of that of the control cells on the non-toxic control surface. mRNA levels of IL-6 and VEGFA correlated well with each other. CONCLUSION: Except for higher viability of cells cultured on polished zirconia specimens, no universally applicable advantage could be found for the ultrasonic cleaning procedure for zirconia and titanium abutment surfaces regarding cell viability, IL-6 expression or VEGFA expression. The cleaning procedures did not have any negative effect either.

Marginal and Internal Adaptation of Hybrid Abutment Assemblies After Central and Local Manufacturing, Respectively.

PURPOSE: Central manufacturing of two-piece computer-aided design/computer-aided manufacture (CAD/CAM) zirconia abutments may provide a higher accuracy of internal and external adaptation at the expense of delayed restoration delivery. The aim of this study was to compare the fit of two-part zirconia abutments that were either fabricated centrally with the DEDICAM system or at a local laboratory. The field of interest was the marginal, external, and internal luting gap between the titanium insert and CAD/CAM zirconia coping. MATERIALS AND METHODS: Two groups of nine two-piece CAD/CAM zirconia hybrid abutments were subjected to scanning electron microscopy (SEM) to evaluate the precision of fit and thickness of the adhesive joint. Control specimens were fabricated with the CAMLOG DEDICAM system at the manufacturer's site; the test specimens were produced in a local laboratory. After embedding all samples (n = 18) in resin, they were sectioned, and the external, marginal, and internal luting gaps between the titanium base and zirconia coping were measured with SEM. Welch's t test was used for statistical analysis of the obtained data. RESULTS: The overall range of measured gaps between the components of two-piece CAD/CAM zirconia abutments was 0 to 115.5 µm; the mean overall gap size and standard deviation was 45.61 ± 5.88 µm and showed no appreciable difference between the test and control groups. The mean sizes of the marginal/external and internal gaps showed only negligible differences. The internal gap size was generally larger and showed a higher variability than the marginal/external gaps, albeit on a very low level. None of the reported differences between the test and control specimens were statistically significant. CONCLUSION: Luting-gap sizes of CAMLOG DEDICAM- and locally fabricated CAD/CAM zirconia hybrid abutments showed no appreciable difference. Both configurations of two-piece abutments provided a highly precise fit of hybrid components, overmatching the high-quality standards in CAD/CAM implant-based prosthetic dentistry.

Influence of scaling procedures on the integrity of titanium nitride coated CAD/CAM abutments.

PURPOSE: To determine the extent of treatment traces, the roughness depth, and the quantity of titanium nitride (TiN) removed from the surface of CAD/CAM abutments after treatment with various instruments. MATERIALS AND METHODS: Twelve TiN coated CAD/CAM abutments were investigated for an in vitro study. In the test group (9), each abutment surface was subjected twice (150 g vs. 200 g pressure) to standardized treatment in a simulated prophylaxis measure with the following instruments: acrylic scaler, titanium curette, and ultrasonic scaler with steel tip. Three abutments were used as control group. Average surface roughness (Sa) and developed interfacial area ratio (Sdr) of treated and untreated surfaces were measured with a profilometer. The extent of treatment traces were analyzed by scanning electron microscopy. RESULTS: Manipulation with ultrasonic scalers resulted in a significant increase of average surface roughness (Sa, P<.05) and developed interfacial area ratio (Sdr, P<.018). Variable contact pressure did not yield any statistically significant difference on Sa-values for all instruments (P=.8). Ultrasonic treatment resulted in pronounced surface traces and partially detachment of the TiN coating. While titanium curettes caused predominantly moderate treatment traces, no traces or detectable substance removal has been determined after manipulation with acrylic curettes. CONCLUSION: Inappropriate instruments during regular plaque control may have an adverse effect on the integrity of the TiN coating of CAD/CAM abutments. To prevent defects and an increased surface roughness at the transmucosal zone of TiN abutments, only acrylic scaling instruments can be recommended for regular maintenance care.

Limitations in molecular detection of lymph node micrometastasis from colorectal cancer.

Colorectal cancer patients with lymph node metastasis have a shorter survival and may require adjuvant therapy after surgery of the primary tumor. It is supposed that a more reliable diagnosis can be achieved using tumor-specific DNA mutations for the detection of metastasizing cells. To design a practical approach for a molecular diagnosis of micrometastasis, we applied direct DNA sequencing to screen 48 early stage colorectal carcinomas for the most frequent mutations of the KRAS, P53, and APC tumor genes. KRAS mutations were detected as frequently as described earlier. In contrast, the frequency of P53 and APC hot spot mutations was unexpectedly low, compared with previous studies using other screening methods or including advanced tumor stages. Not more than 31% of early stage tumors showed a mutation in at least 1 of the selected hot spot codons. Applying mutant-enriched polymerase chain reaction (PCR), the mutation of the primary tumor was detected in lymph node DNA from 2 of the KRAS-positive patients. In 1 patient, the result was not verified by subtractive iterative PCR, a principally different molecular method with high sensitivity and specificity. Our data suggest that screening for suitable markers for a molecular detection of occult lymph node metastasis cannot be restricted to small-sized hot spot regions of a few tumor genes and possibly must include tumor-specific epigenetic changes. Furthermore, restriction enzyme-based methods such as mutant-enriched PCR are not suitable to detect any mutation with equal efficiency and they should be carefully controlled to avoid false-positive detection of marker mutations in lymph node DNA.

Microscopical and chemical surface characterization of CAD/CAM zircona abutments after different cleaning procedures. A qualitative analysis.

PURPOSE: To describe and characterize the surface topography and cleanliness of CAD/CAM manufactured zirconia abutments after steaming and ultrasonic cleaning. MATERIALS AND METHODS: A total of 12 ceramic CAD/CAM implant abutments of various manufacturers were produced and randomly divided into two groups of six samples each (control and test group). Four two-piece hybrid abutments and two one-piece abutments made of zirconium-dioxide were assessed per each group. In the control group, cleaning by steam was performed. The test group underwent an ultrasonic cleaning procedure with acetone, ethyl alcohol and antibacterial solution. Groups were subjected to scanning electron microscope (SEM) analysis and Energy-dispersive X-ray spectroscopy (EDX) to verify and characterize contaminant chemical characterization non-quantitatively. RESULTS: All zirconia CAD/CAM abutments in the present study displayed production-induced wear particles, debris as well as organic and inorganic contaminants. The abutments of the test group showed reduction of surface contamination after undergoing an ultrasonic cleaning procedure. However, an absolute removal of pollutants could not be achieved. CONCLUSION: The presence of debris on the transmucosal surface of CAD/CAM zirconia abutments of various manufacturers was confirmed. Within the limits of the study design, the results suggest that a defined ultrasonic cleaning process can be advantageously employed to reduce such debris, thus, supposedly enhancing soft tissue healing. Although the adverse long-term influence of abutment contamination on the biological stability of peri-implant tissues has been evidenced, a standardized and validated polishing and cleaning protocol still has to be implemented.

In vitro fatigue and fracture resistance of one- and two-piece CAD/CAM zirconia implant abutments.

PURPOSE: All-ceramic abutments are employed increasingly often in implant dentistry for esthetic reasons. In vitro stress testing is required to evaluate the suitability of these constructions, especially in load-bearing posterior regions. The purpose of the study was to assess and compare the fatigue and fracture resistance of one- and two-piece computer-aided design/computer-assisted manufacture (CAD/CAM) zirconia implant abutments with an internal-hex connection and prefabricated commercially available zirconia stock abutments. MATERIALS AND METHODS: Twenty-one abutment-crown specimens were prepared for three test groups. Control group 1 (SZ) included specimens with unprepared stock zirconia abutments, test group 2 (OP) included one-piece CAD/CAM zirconia abutments, and test group 3 (TP) included two-piece CAD/CAM zirconia abutments. All 21 specimens underwent thermocycling and fatigue testing. Finally, all specimens were tested for fracture resistance with a universal testing machine. The maximum load was applied to the tapered occlusal area of each crown at a 30-degree angle and a crosshead speed of 0.5 mm/min until the implant-abutment connection failed. Kolmogorov-Smirnov, Shapiro-Wilk, and post-hoc Scheffé tests were used for statistical analysis. RESULTS: All abutments in groups SZ and OP fractured into two or more pieces after fracture resistance testing. None of the TP abutments displayed apparent disintegration, but failure was evidenced by bending of the retention screw. OP abutments (232.1 ± 29.8 N) and SZ abutments (251.8 ± 23.2 N) showed lower fracture loads than the TP abutments (291.4 ± 27.8 N). However, only the difference between the OP and TP groups was statistically significant. Further load-displacement analyses corroborated the higher mechanical stability of the TP abutments. CONCLUSION: Superior resistance was achieved for two-piece hybrid CAD/CAM zirconia abutments. These abutments might be clinically beneficial in high-load areas, such as premolar and molar regions.

Nucleic acids from intact epithelial cells as a target for stool-based molecular diagnosis of colorectal cancer.

Stool-based molecular techniques may improve strategies for colorectal cancer screening. Molecular methods have successfully been applied to detect tumour DNA in stool from patients diagnosed for colorectal carcinoma. In these assays human DNA has to be analyzed against a background of excess nucleic acids from bacteria and dietary waste products. More recently a different diagnostic approach has been described characterizing intact cells isolated from stool. In this study we combine both approaches preparing nucleic acids from isolated epithelial cells to evaluate if: a) tumour cell-specific RNA can be analyzed since cellular RNA molecules are prevented from early digestion by an intact cell membrane; and b) specificity or sensitivity of established DNA-based methods can be improved when epithelial cells are separated from other stool components. Comparing different protocols we found cell isolation using epithelium-specific antibodies to be more effective and reproducible than a technique using density gradient centrifugation. A detection limit of 10(4) cells per ml stool was determined when samples from healthy volunteers were spiked with epithelial cells. Amplification of human sequences from total stool DNA was more efficient than a correspondent amplification of DNA extracted from isolated cells, so that an improvement of DNA-based methods cannot be expected by introducing cell isolation procedures. RNA detection was successful in 1 of 5 patients with confirmed diagnosis of colorectal cancer. The authors suggest that low numbers of detectable cells might rather be a biological than an analytical problem limiting a routinely performed method for colorectal cancer diagnosis.

Retentive strength of two-piece CAD/CAM zirconia implant abutments.

PURPOSE: The purpose of this study is to evaluate the retention of two-piece computer-aided design (CAD)/computer aided manufacturing (CAM) zirconia abutments after artificial aging under simulated oral conditions using three different types of resin-based luting agents. MATERIAL AND METHODS: Twenty-one CAD/CAM-generated zirconia copings (CERCON Compartis, Degudent, Hanau, Germany) were bonded to a prefabricated secondary titanium implant insert (XiVE Ti-Base, Dentsply Friadent, Mannheim, Germany), using three different types of resin-based luting agents: group A: Panavia 21 (Kuraray Co, Kurashiki, Japan); group B: Multilink Implant (Ivoclar Vivadent, Schaan, Liechtenstein); and group C: SmartCem2 (Dentsply DeTrey, Konstanz, Germany). The bonding surfaces of the titanium inserts and the zirconia ceramic copings were air-abraded and cleaned in alcohol. All specimens were stored in distilled water for 60 days and subsequently thermal-cycled 15,000 times (5-55 °C). The dislodging force of the copings along the long axis of the implant/abutment complex was recorded using a universal testing machine with 2 mm/min crosshead speed. Data were analyzed descriptively and by performing the Kruskal-Wallis test. RESULTS: The mean retention values were 924.93 ± 363.31 N for Panavia 21, 878.05 ± 208.33 N for Multilink Implant, and 650.77 ± 174.92 N for SmartCem2. The Kruskal-Wallis test indicated no significant difference between the retention values of the tested luting agents (p = 0.1314). The failure modes of all tested two-piece abutments were completely adhesive, leaving the detached zirconia coping and titanium insert undamaged. CONCLUSION: The use of resin-based luting agents in combination with air abrasion of titanium inserts and zirconia copings led to a stable retention of two-piece CAD/CAM abutments. The bonding stability of the investigated luting agents exceeded the general limits of fracture resistance of two-piece zirconia abutments. A notable difference between the mean retention values of the tested bond materials was shown. However, the statistical analysis revealed that this difference was not significant.

Range of indications for translucent zirconia modifications: clinical and technical aspects.

Translucent zirconia modifications offer esthetic improvement for manually veneered zirconia structures, as they do not lead to a shining through of the substructure material, even in cases with a pronounced anatomic core design for maximum support of the veneering ceramics. Moreover, these zirconia modifications allow the production of fully anatomic zirconia crowns and fixed dental prostheses in the posterior region. The clinical advantage of these restorations is defined by a significantly reduced material thickness in comparison with veneered restorations or other monolithic materials. As the restoration can be colored individually prior to sintering, followed by characterization by staining, good esthetic results in the posterior region are achieved, even in cases with substantially reduced space. The results of laboratory studies performed so far seem to justify the clinical application of fully anatomic restorations. However, additional clinical studies are required to support these new material modifications.

Release kinetics of VEGF165 from a collagen matrix and structural matrix changes in a circulation model.

BACKGROUND: Current approaches in bone regeneration combine osteoconductive scaffolds with bioactive cytokines like BMP or VEGF. The idea of our in-vitro trial was to apply VEGF165 in gradient concentrations to an equine collagen carrier and to study pharmacological and morphological characteristics of the complex in a circulation model. METHODS: Release kinetics of VEGF165 complexed in different quantities in a collagen matrix were determined in a circulation model by quantifying protein concentration with ELISA over a period of 5 days. The structural changes of the collagen matrix were assessed with light microscopy, native scanning electron microscopy (SEM) as well as with immuno-gold-labelling technique in scanning and transmission electron microscopy (TEM). RESULTS: We established a biological half-life for VEGF165 of 90 minutes. In a half-logarithmic presentation the VEGF165 release showed a linear declining gradient; the release kinetics were not depending on VEGF165 concentrations. After 12 hours VEGF release reached a plateau, after 48 hours VEGF165 was no longer detectable in the complexes charged with lower doses, but still measurable in the 80 microg sample. At the beginning of the study a smear layer was visible on the surface of the complex. After the wash out of the protein in the first days the natural structure of the collagen appeared and did not change over the test period. CONCLUSIONS: By defining the pharmacological and morphological profile of a cytokine collagen complex in a circulation model our data paves the way for further in-vivo studies where additional biological side effects will have to be considered. VEGF165 linked to collagen fibrils shows its improved stability in direct electron microscopic imaging as well as in prolonged release from the matrix. Our in-vitro trial substantiates the position of cytokine collagen complexes as innovative and effective treatment tools in regenerative medicine and and may initiate further clinical research.