Real-time co-site optical microscopy study on the morphological changes of the dentine's surface after citric acid and sodium hypochlorite: a single-tooth model.

BACKGROUND: To this day, the effects of sodium hypochlorite and chelating agents on the smear layer and on the dentine's surface, remain not fully examined. The study is aimed to analyze the dentine's surface treated with 40% citric acid and 5.25% sodium hypochlorite according to two irrigation protocols. MATERIALS AND METHODS: The study employed a computer-controlled Nikon Eclipse LV100 optical microscope. Ten roots split longitudinally with canals prepared mechanically using the MTwo system to a size of 40/04 were observed. The root halves were divided into two study groups, one half in each of the groups. According to two irrigation protocols, the dentine's surface was irrigated with 40% citric acid and 5.25% sodium hypochlorite, separated with water. Dentine surface was observed in a fixed place and photographed in 500 × magnification after each irrigation stage. The obtained images were then analyzed using computer software (NIS-Elements AR, GIMP-2.6, ImageJ 1.45s). RESULTS: Various speed of removal of the smear layer and varied morphological changes of the dentine's surface were observed in both examined groups. Double irrigation of the dentine with citric acid for 30 s resulted in complete removal of the smear layer, and double irrigation for 60 s resulted in increased mean diameter of the dentine tubules (degradation of the dentine's surface). CONCLUSIONS: Double alternating irrigation with citric acid and sodium hypochlorite for 30 s yielded satisfactory results, while double irrigation for 60 s resulted in increased mean diameter of the dentine tubules. A real time assessment of the dentine's surface changes after both tested solutions can improve understanding of the phenomena that occur during the irrigation and as a result it might help to improve clinical outcomes in root canal treatment.

Direct tool for quantitative analysis of cell/object dynamic behavior - metastasis and far beyond.

INTRODUCTION: The dynamics and depth of invasion as well as the ability of cancer cells to penetrate the walls of  lymphatic or blood vessels represent critical survival-influencing factors in cancer patients. Depending on the cell type and tissue environment, cancer cell invasion differ in terms of motility mechanism and migration modes. Thus, there is the need of effective models allowing not only for single cell invasion potential assessment but also for collective migration and expansive growth evaluation in 3D microenvironment e.g. basement membranes. To meet this task, the specimens should be compared and analyzed in terms of the dynamics of movement and the evolution of the shape. OBJECTIVES: Our main objective was development of the mathematical method that enables fast and credible calculation of parameters of shape and position, namely standard deviations (σX, σY), centroid position (µX, µY) and correlation coefficient ρ, based only on the contour of the aggregate. METHODS: In order to accomplish this goal we measured geometrical properties of aggregates of RGM1 cells seeded in 3D Geltrex basement membrane. Referential microscopic images were taken 24 and 48 h after seeding and cell group dynamics was registered over 8 h periods using time lapse microscopy. RESULTS: Based on gathered data, we managed to develop and fully test universal numerical tool allowing for estimation of statistical parameters of cell groups and aggregates which then allows for the precise evaluation of their behavior within microenvironment with time. CONCLUSION: We conclude, that our tool is suitable for any research on the metastatic potential and motility of cancer cells in a given microenvironment, regardless of the migration mechanism, which together with the advanced analysis like cell single-cell transcriptomic, proteomic, and chromatin accessibility data may allow to identify precise targets for anti-cancer therapies, to predict the degree of malignancy of neoplastic lesions as well as it can be useful during architecting therapeutic strategies. Moreover, the developed tool seems to be broadly applicable for assessment of behavioural dynamics of any population.

Effect of sodium hypochlorite, isopropyl alcohol and chlorhexidine on the epoxy sealant penetration into the dentinal tubules.

BACKGROUND: The sealer penetration into the dentinal tubules might be beneficial, especially in necrotic endodontic cases, as it provides the obstruction of the contaminated tubules. OBJECTIVES: To determine the effect of 3 final irrigants (sodium hypochlorite (NaOCl), alcohol and chlorhexidine (CHX)) on the penetration of an epoxy sealer into the dentinal tubules. MATERIAL AND METHODS: The study was carried out on 60 single-canal human teeth with straight roots. The root canals were prepared to the ISO 40/04 size, using the Reciproc® instruments. The teeth were divided into 4 groups (n = 15). The canals in each group were irrigated according to the following scheme: group 1 (control) - 5.25% NaOCl; group 2 - smear layer removal (40% citric acid (CA) and 5.25% NaOCl) and 5.25% NaOCl; group 3 - smear layer removal (as in group 2), and 40% CA, water and 98% isopropyl alcohol; and group 4 - smear layer removal (as in group 2), and 40% CA, water and 2% CHX. The root canals were filled using the vertical condensation technique with gutta-percha and the porphyrin-labeled AH Plus™ sealer. After 3 days, 1-milimeter-thick cross-section slices were cut from the roots at a distance of 2 mm, 5 mm and 8 mm from the apex. The sections were imaged under a confocal microscope and the sealant penetration depth into the dentinal tubules was measured. RESULTS: The longest resin tags in all parts of the roots were found in group 4 (CHX), and the shortest in group 1 (control). The mean depth of the sealer penetration (in micrometers) was as follows: 21, 22 and 23 (group 1); 201, 231 and 374 (group 2); 170, 232 and 280 (group 3); and 330, 408 and 638 (group 4) in the apical, middle and coronal parts, respectively. CONCLUSIONS: The final irrigation with CHX resulted in the deepest penetration of the epoxy sealer into the tubules. Isopropyl alcohol had the most negative impact on the sealer penetration into the tubules.

The impact of irrigation protocols on epoxy sealer penetration depth in dentinal tubules. Study involving laser confocal microscopy.

The aim was to assess the impact of irrigation protocols ended with ethanol or chlorhexidine on AH Plus penetration into dentinal tubules. 45 root canals were prepared to ISO 40/04, divided into three groups and irrigated with three protocols: Group 1 (control): 5.25% NaOCl; Group 2: 40% CA (citric acid), 5.25% NaOCl, 40% CA, water, ethanol; Group 3: 40% CA, 5.25% NaOCl, 40% CA, water, 2% chlorhexidine. Canals were filled using vertical condensation technique with gutta-percha and fluorescein-stained AH Plus sealer. After 72 h, 1 mm thick cross-sections were cut at 2, 5, 8 mm from the apex. Confocal laser microscope was used to measure the sealer penetration into dentinal tubules. Mean depth of sealer penetration (in micrometres) was 107, 131, 170 (Group 1); 146, 233, 317 (Group 2); 185, 301, 542 (Group 3); in apical, middle and coronal parts, respectively. Irrigation protocol ended with chlorhexidine resulted in the deeper sealer penetration compared with alcohol.

Time-extended exposure of gastric epithelial cells to secretome of Helicobacter pylori-activated fibroblasts induces reprogramming of gastric epithelium towards pre-cancerogenic and pro-invasive phenotype.

Despite of the improvement in gastric cancer (GC) therapies patients still suffer from cancer recurrence and metastasis. Recently, the high ratio of these events combined with increased chemoresistance has been related to the asymptomatic Helicobacter pylori (Hp) infections. The limited efficiency of GC treatment strategies is also increasingly attributed to the activity of tumor stroma with the key role of cancer-associated fibroblasts (CAFs). In order to investigate the influence of Hp infection within stromal gastric tissue on cancer initiation and progression, we have exposed normal gastric epithelial cells to long-term influence of Hp-activated gastric fibroblast secretome. We have referred obtained results to this secretome influence on cancer cell lines. The invasive properties of cells were checked by time-lapse video microscopy and basement membrane assays. The expression of invasion-related factors was checked by RT-PCR, Western Blot, immunofluorescence and Elisa. Hp-activated gastric fibroblast secretome induced EMT type 3-related shifts of RGM1 cell phenotype; in particular it augmented their motility, cytoskeletal plasticity and invasiveness. These effects were accompanied by Snail1/Twist activation, the up-regulation of cytokeratin19/FAP/TNC/Integrin-ß1 and MMPs, and by the induction of cMethigh/pEGFRhigh phenotype. Mechanistic studies suggest that this microevolution next to TGFß relies also on c-Met/EGFR signaling interplay and engages HGF-Integrin-Ras-dependent Twist activation leading to MMP and TNC upregulation with subsequent positive auto- and paracrine feedback loops intensifying this process. Similar shifts were detected in cancer cells exposed to this secretome. Collectively, we show that the secretome of Hp-infected fibroblasts induces reprogramming/microevolution of epithelial and cancer cells towards type 3 EMT-related invasive phenotype in a manner reciprocally reliant next to TGFß on cMet/Integrin-ß1/p-EGFR-dependent axis. Apparently, the phenotypical plasticity of Hp-activated fibroblast reprogrammed gastric epithelial cells determines their susceptibility to the pro-invasive signaling, which results in re-organization of gastric niches and provides the cues for GC promotion/progression.

[The impact of polychlorinated biphenyls on placental and ovarian steroidogenesis]. / Polichlorowane bifenyle, a sterydogeneza w lozysku i jajniku.

The structure and turnover of polichlorinated biphenyls (PCB) in environment was presented. The PCB absorption and metabolism in organisms as well as the PCB induced disruption of placental and ovarian steroidogenesis were discussed.

[The impact of polichlorinated biphenyls on neoplastic processes]. / Wplyw polichlorowanych bifenyli na procesy nowotworzenia.

The structure and turnover of polichlorinated biphenyls (PCB) in environment was presented. The PCB absorption and metabolism in organisms as well as the PCB influence on neoplastic process.

The effects of alternate irrigation of root canals with chelating agents and sodium hypochlorite on the effectiveness of smear layer removal.

BACKGROUND: After the mechanical preparation of a root canal, the canal walls are covered with a smear layer. In order to deeply clean the dentinal tubules, removal of the smear layer is recommended. There is no consensus on the length of time of rinsing with chelating agents or irrigation with alternating chelating agents and sodium hypochlorite (NaOCl). OBJECTIVES: The aim of the study was to evaluate the effectiveness of smear layer removal using 4 irrigation protocols. MATERIAL AND METHODS: We prepared 42 straight root canals to size ISO40/04 and assigned them into 4 study groups (n = 10) and a control group (n = 2). The root canals were irrigated as follows: in the control group, 180 s with 5.25% NaOCl; in group 1, 60 s with 40% citric acid (CA) and 120 s with NaOCl; in group 2, 120 s with CA and 120 s with NaOCl; in group 3, 30 s CA, 30 s with NaOCl, 30 s CA and 120 s with NaOCl; and in group 4, 60 s with CA, 30 s with NaOCl, 60 s with CA, and 120 s with NaOCl. The roots were split longitudinally and the root canals were observed under ×200-500 magnification. The root canal walls were analyzed in areas 2 mm, 6 mm and 10 mm from the apex. RESULTS: In the apical and medial sections, the best effects were achieved in groups 3 and 4. In coronal sections, no significant differences between experimental groups were found. CONCLUSIONS: Within the limitations of this study, it can be concluded that irrigation with alternating NaOCl and CA was the most effective at smear layer removal, regardless of the irrigation time.