Pre-treatment of cattle sperm and/or oocyte with antibody to lipocalin type prostaglandin D synthase inhibits in vitro fertilization and increases sperm-oocyte binding

The present study was conducted to determine the affect of pre-treating of oocytes and/or sperm with a rabbit polyclonal antibody against recombinant cattle lipocalin type prostaglandin D synthase (alpha L-PGDS) on in vitro sperm-oocyte binding and fertilization. In vitro matured cattle oocytes were incubated (39 degrees C, 5% CO2 in air) for I It in the following treatments either 500 mu L of fertilization medium (FM) or FM with alpha L-PGDS (1:2000). Frozen-thawed spermatozoa were washed by a 45/90% layered Percoll gradient centrifugation and incubated for I h either FM or FM with a L-PGDS. This study utilized five different treatments: (1) no antibody (control); (2) a rabbit IgG against a non-bovine antigen, bacterial histidase (alpha-hist); (3) a L-PGDS at fertilization time (with fertilization medium); (4) alpha L-PGDS-treated oocytes; or (5) a L-PGDS-treated sperm. Pre-treated oocytes were incubated with 10 X 10(4) washed spermatozoa per 25 oocytes. Oocytes used to assess sperm binding were stained with Hoescht 33342, and the number of sperm bound per zonae pellucidae counted. The remaining oocytes were fixed in acid alcohol, stained with 1% acetate-orcein and observed to determine the presence of pronuclei. More sperm bound to the zonae pellucidae when oocytes and/or sperm were pre-treated with alpha. L-PGDS: (1) 26.4 +/- 3.0; (2) 25.6 +/- 3.0; (3) 59.7 +/- 3.0; (4) 56.4 +/- 3.0; and (5) 57.1 +/- 3.0. Addition of alpha L-PGDS with sperm, oocytes, or both, decreased fertilization (P < 0.05) compared with the control: (1) 89.2 +/- 2.0%; (2) 87.5 +/- 2.0%; (3) 19.4 +/- 2.0%; (4) 27.2 +/- 3.1%; and (5) 14.1 +/- 3.4%. The alpha L-PGDS reacts with both oocytes and spermatozoa, resulting in increases of in vitro sperm-oocyte binding and inhibition of fertilization. These observations suggest that L-PGDS may have a role in cattle fertilization. (c) 2007 Elsevier B.V. All rights reserved.

Evaluation of three different adhesive systems using a bacterial method to develop secondary caries in vitro

Purpose: To assess the effects of three different dental adhesive systems on the formation of secondary root caries, in vitro, with a standardized interfacial gap in a filled cavity model. Methods: 40 sound human molars were selected and randomly assigned to four experimental groups: Clearfil SE Bond (CSEB), Xeno III (X-III), Scotchbond Multi-Purpose Plus (SBMP) and negative control (NC) without an adhesive system. After the standardized Class V cavity preparations on the buccal and lingual surfaces, restorations were placed with resin composite (Filtek Z250) using a standardized interfacial gap, using a 3 x 2 mm piece of 50 mu m metal matrix. The teeth were sterilized with gamma irradiation and exposed to a cariogenic challenge using a bacterial system with Streptococcus mutans. Depth and extension of wall lesions formed and the depth of outer lesions were measured by software coupled with light microscopy. Results: For wall lesion extension the ANOVA test showed differences between groups except between X-HI and SBMP (P= 0.294). The Tukey`s test of confidence intervals indicated smaller values for the CSEB group than for the others. For wall lesion depth the CSEB group also presented the smallest mean values of wall lesion depth when compared to the others (P< 0.0001) for all comparisons using Tukey`s test. Regarding outer lesion depth, all adhesives showed statistically similar behavior. SEM evaluation of the morphologic appearance of caries lesions confirmed the statistical results showing small caries lesion development for cavities restored with CSEB adhesive system, which may suggest that this adhesive system interdiffusion zone promoted a good interaction with subjacent dentin protecting the dental tissues from recurrent caries. (Am J Dent 2010;23:93-97).

Bacterial Adhesion on Smooth and Rough Titanium Surfaces After Treatment With Different Instruments

Background: Newly formed biofilm after implant debridement may challenge the long-term stability of peri-implant therapy. This in vitro study aimed to assess the roughness and adherence of Streptococcus sanguinis after treatment of smooth and rough titanium surfaces with an erbium-doped: yttrium, aluminum, and garnet (Er:YAG) laser, metal and plastic curets, and an air-powder abrasive system. Methods: Forty titanium disks with smooth-machined surfaces and 40 with sand-blasted and acid-etched surfaces were divided into the following treatment groups: Er:YAG laser; plastic curet; metal curet, and air-powder abrasive system. The surface roughness (roughness average [Raj) before and after treatments was determined using a profilometer. S. sanguinis (American Type Culture Collection 10556) was grown on treated and untreated specimens, and the amounts of retained bacteria on the surfaces were measured by the culture method. Rough and smooth surfaces with and without a suspension of S. sanguinis were also analyzed using scanning electron microscopy (SEM). Results: For smooth surfaces, the roughest surfaces were produced by metal curets (repeated - measures analysis of variance [ANOVA] and Tukey test; P<0.05). The rough-surface profile was not altered by any of the treatments (repeated-measures ANOVA; P>0.05). Rough surfaces treated with metal curets and air-powder abrasion showed the lowest level of bacteria] adhesion (two-way ANOVA and Tukey test; P<0.05). SEM analysis revealed distinct surface profiles produced by all devices. Conclusions: Metal curets are not recommended for smooth titanium surface debridement due to severe texture alteration. Rough surfaces treated with a metal curet and the air-powder abrasive system were less susceptible to bacterial adhesion, probably due to texture modification and the presence of abrasive deposits. J Periodontol 2009;80: 1824-1832.

Immunoexpression of Ki67, proliferative cell nuclear antigen, and Bcl-2 proteins in a case of ameloblastic fibrosarcoma

Ameloblastic fibrosarcoma (AFS), regarded as the malignant counterpart of the benign ameloblastic fibroma, is an extremely rare odontogenic neoplasm with only 68 cases reported in the English literature up to 2009. It is composed of a benign odontogenic epithelium, resembling that of ameloblastoma, and a malignant mesenchymal part exhibiting features of fibrosarcoma. Due to the rarity of the lesion, little is known about its molecular pathogenesis; therefore, in the current study, we sought to evaluate the immunoexpression of Ki67, proliferative cell nuclear antigen, and Bcl-2 proteins in AFS, comparing the results obtained with its benign counterpart, as well as to report a new case of this rare entity affecting a 19-year-old female patient. The results obtained revealed that all the proteins evaluated were overexpressed in the malignant mesenchymal portion of AFS if compared with ameloblastic fibroma, suggesting that nuclear proliferative factors such as Ki67 and proliferative cell nuclear antigen, in association to histopathologic features, may be useful markers for identifying the malignancy and that, despite the lack of molecular analysis in the case reported, Bcl-2 alteration may play a role in AFS pathogenesis. (C) 2010 Elsevier Inc. All rights reserved.

Bacterial leakage in obturated root canals-part 2: a comparative histologic and microbiologic analyses

Objective. In this study, presence of dentin infection in root canals, obturated with 4 techniques submitted to the bacterial leakage test, was evaluated using histologic methods. Study design. The canals of palatal roots of 160 molars were instrumented and divided into different groups, according to the obturation technique used (lateral condensation, MicroSeal system, Touch `n Heat + Ultrafil, and Tagger`s hybrid technique) and extent of the remaining obturation material (5 mm and 10 mm). Ten additional roots were used as control samples. The roots were sterilized in ethylene oxide and mounted on a device for evaluation of bacterial leakage using the bacteria Enterococcus faecalis for 120 days. After the leakage test, roots were microscopically analyzed for the presence of dentin infection in the root canals and dentinal tubules. Results. A total of 154 specimens were analyzed using both methodologies in the experimental groups; 50 root canals (32.4%) showed bacterial leakage at the end of the experimental period, and 118 (76.6%) showed the presence of bacteria in the root canals using the histologic criteria. The lateral condensation technique allowed lower penetration of bacteria in the root canals and dentinal tubules, followed by Touch `n Heat + Ultrafil, MicroSeal, and Tagger`s hybrid technique, which allowed significantly greater penetration of bacteria. Root canals with 10 mm of remaining obturation material presented similar bacterial penetration as root canals with 5 mm. Conclusions. Even when an adequate seal of the apical foramen was shown by the absence of turbidity in the bacterial leakage test, E. faecalis dentin infection was present in a high percentage of the root canals after 120 days of root filling exposure to the bacteria. Tagger`s hybrid technique presented greater quantity of bacteria in histologic sections than root canals obturated with the other techniques. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010; 109: 788-794)

Bacterial leakage in root canals obturated by different techniques. Part 1: microbiologic evaluation

Objective. This study compared the coronal bacterial leakage of root canals obturated by different techniques and with different lengths of obturation. Study design. The canals of palatal roots of 160 maxillary molars were instrumented and divided into different groups according to the obturation technique used (lateral condensation, Microseal system, Touch `n Heat + Ultrafil system, or Tagger`s hybrid technique) and the length of obturation (5 mm or 10 mm). The roots were impermeabilized, sterilized in ethylene oxide, and mounted on a device for evaluation of the bacterial leakage. Results. Tagger`s hybrid technique produced a statistically greater number of specimens with coronal leakage than the other techniques. There was no statistically significant difference between the lateral condensation, Touch `n Heat + Ultrafil, and Microseal groups. Root canals with 10 mm of obturation produced a statistically significantly smaller number of specimens with leakage than root canals with 5 mm of obturation. Conclusion. Tagger`s hybrid technique produced a greater number of specimens with coronal leakage than the other techniques, and a greater number of root canals with 5 mm of obturation leaked than root canals with 10 mm of obturation.

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)

In Vitro Evaluation of Bacterial Leakage Along the Implant-Abutment Interface of an External-Hex Implant After Saliva Incubation

The aim of this in vitro study was to evaluate bacterial leakage along the implant-abutment interface under unloaded conditions. Twelve premachined abutments with plastic sleeves and 12 dental implants were used in this study. Prior to tests of bacterial leakage, samples from the inner parts of the implants were collected with sterile microbrushes to serve as negative controls for contamination. After casting, the abutments were tightened to 32 Ncm on the implants. The assemblies were immersed in 2.0 mL of human saliva and incubated for 7 days. After this period, possible contamination of the internal parts of the implants was evaluated using the DNA Checkerboard method. Microorganisms were found in the internal surfaces of all the implants evaluated. Aggregatibacter actinomycetemcomitans and Capnocytophaga gingivalis were the most incident species. No microorganisms were found in the samples recovered from the implants before contamination testing (negative control). Bacterial species from human saliva may penetrate the implant-abutment interface under unloaded conditions. INT J ORAL MAXILLOFAC IMPLANTS 2011;26:782-787

Influence of repeated screw tightening on bacterial leakage along the implant-abutment interface

Objectives Bacterial penetration along the implant-abutment interface as a consequence of abutment screw loosening has been reported in a number of recent studies. The aim of this in vitro study was to investigate the influence of repeated tightening of the abutment screw on leakage of Streptococcus mutans along the interface between implants and pre-machined abutments. Materials and methods Twenty pre-machined abutments with a plastic sleeve were used. The abutment screws were tightened to 32 N cm in group 1 (n=10 - control) and to 32 N cm, loosened and re-tightened with the same torque twice in group 2 (n=10). The assemblies were completely immersed in 5 ml of Tryptic Soy Broth medium inoculated with S. mutans and incubated for 14 days. After this period, contamination of the implant internal threaded chamber was evaluated using the DNA Checkerboard method. Results Microorganisms were found on the internal surfaces of both groups evaluated. However, bacterial counts in group 2 were significantly higher than that in the control group (P < 0.05). Conclusion These results suggest that bacterial leakage between implants and abutments occurs even under unloaded conditions and at a higher intensity when the abutment screw is tightened and loosened repeatedly. To cite this article:do Nascimento C, Pedrazzi V, Kirsten Miani P, Daher Moreira L, de Albuquerque Junior RF. Influence of repeated screw tightening on bacterial leakage along the implant-abutment interface.Clin. Oral Impl. Res. 20, 2009; 1394-1397.doi: 10.1111/j.1600-0501.2009.01769.x.