Sibel Cetik

Comparison of GLUT1, GLUT2, GLUT4 and SGLT1 mRNA Expression in the Salivary Glands and Six Other Organs of Control, Streptozotocin-Induced and Goto-Kakizaki Diabetic Rats

Background/Aims: The expression and localization of several distinct glucose transporters (GLUT1, GLUT2, GLUT4, and SGLT1) was recently characterized in the parotid gland of normal rats by quantitative real-time PCR analysis, immunohistochemistry and Western blotting. The major aims of the present study was to compare the mRNA expression of these glucose transporters in both the parotid gland and submaxillary gland of control rats, streptozotocin-induced diabetic rats and hereditarily diabetic Goto-Kakizaki rats. Methods: Quantitative real-time PCR analysis was performed in the parotid and submaxillary salivary glands and, for purpose of comparison, also in the heart, kidney, liver, lung, muscle and pancreas from control animals and either streptozotocin-treated or Goto-Kakizaki rats. Results: The expression of GLUT4, but not GLUT1 or SGLT1, mRNA was decreased in the diabetic rats. The results also allow comparing both the mRNA expression level of the four glucose transporters in salivary glands and six other organs, and the diabetes-induced changes in such an expression in distinct locations. Conclusion: The mRNA expression of the insulin-dependent GLUT4 transporter was the sole to be significantly decreased in the salivary glands of diabetic animals. The possible consequence of such a decrease in terms of the control of salivary glucose concentration requires further investigation.

Glucose Transport by Acinar Cells in Rat Parotid Glands

Background/Aims: Salivary glucose is often considered as being from glandular origin. Little information is available, however, on the possible role of glucose transporters in the secretion of the hexose by salivary glands. The major aim of the present study was to investigate the expression and localization of several distinct glucose transporters in acinar cells of rat parotid glands. Methods: Quantitative real-time PCR analysis, immunohistochemistry and western blotting techniques were used to assess the presence of SGLT1, GLUT1, GLUT2 and GLUT4 in acinar cells of rat parotid glands. Results: Quantitative real-time PCR documented the expression of SGLT1 and GLUT1 in parotid tissues, with a much lower level of GLUT4 mRNA and no expression of GLUT2 mRNA. Western blot analysis revealed the presence of SGLT1, GLUT1 and GLUT4 proteins, but not GLUT2 proteins in the parotid extract. Immunohistochemistry confirmed these findings. SGLT1 was specifically located at the baso-lateral membrane, co-localizing with Na+/K+ ATPase. GLUT1 was found both at the baso-lateral and apical level. GLUT4 appeared to be also located at the baso-lateral level. However, too little GLUT4 was present to allow co-localization labeling. Conclusion: Based on these findings, a model is proposed for the transport of glucose into the acinar cells and thereafter into the acinar lumen.

Expression and Localization of Glucose Transporters in Rodent Submandibular Salivary Glands

Background/Aim: The submandibular gland is one of the three major salivary glands, producing a mixed secretion; this saliva is hypotonic compared to plasma. It also secretes glucose, but the mechanisms responsible for this process are poorly understood. Our study addressed the question whether glucose transporters are expressed and how are they localized within specific rodent submandibular cells, in order to estimate a possible implication in salivary glucose disposal. Methods: Immunohistochemistry, RT-qPCR and Western blotting were performed to determine the presence/localization of glucose transporters in rodent submandibular glands. Results: GLUT4 was identified in the submandibular salivary gland at both mRNA and protein level. The immunohistochemical analysis revealed its localization preponderantly in the ductal cells of the gland, near to the basolateral. SGLT1 and GLUT1 were highly expressed in submandibular tissues in both acinar and ductal cells, but not GLUT2. These results were confirmed by RT-qPCR. It was also documented that insulin stimulates the net uptake of D-glucose by ductal rings prepared from submandibulary salivary glands, the relative magnitude of such an enhancing action being comparable to that found in hemidiaphragms. Conclusion: At least three major glucose transporters are expressed in the rodent submandibular glands, of which GLUT4 is specifically localized near the basolateral side of ductal structures. This points-out its possible role in regulating glucose uptake from the bloodstream, most likely to sustain ductal cellular metabolism.

A tentative model for D-glucose turnover in human saliva

OBJECTIVE The aim of the present study is to propose a tentative model for d-glucose turnover in human saliva. The whole saliva and the saliva from parotid and submandibular/sublingual glands were collected by use of the Salivette™. RESULTS The saliva glucose concentration was measured by the hexokinase method, saliva bacteria glycolysis by use of d-[5-(3)H] glucose, and the saliva ATP content by the luciferase method. The concentration of glucose amounted to 43.9±6.3 (n=29), 197.5±17.3 (n=29), 104.0±12.4 (n=27) μM in whole saliva, parotid saliva and submandibular/sublingual saliva, respectively. The rate of d-glucose utilization by oral bacteria at a physiological concentration of d-glucose in saliva (50μM) was estimated at 0.047±0.003 (n=11) nmol/min per 10(6) bacteria. Unstimulated salivary d-glucose turnover rate, as calculated from the amount of glucose secreted in saliva which comes from parotid and submandibular and sublingual glands represented 214.6±19.1%/min. In order for salivary d-glucose production to match bacterial utilization of the hexose, the total number of oral bacteria was estimated at about 2.0×10(9) bacteria, in fair agreement with previously published data. CONCLUSION This study thus provides support for a tentative model for d-glucose turnover in human saliva.

A Comparative Study of Microleakage on Dental Surfaces Bonded with Three Self-Etch Adhesive Systems Treated with the Er:YAG Laser and Bur

Aim. This study sought to compare the microleakage of three adhesive systems in the context of Erbium-YAG laser and diamond bur cavity procedures. Cavities were restored with composite resin. Materials and Methods. Standardized Class V cavities were performed in 72 extracted human teeth by means of diamond burs or Er-YAG laser. The samples were randomly divided into six groups of 12, testing three adhesive systems (Clearfil s3 Bond Plus, Xeno® Select, and Futurabond U) for each method used. Cavities were restored with composite resin before thermocycling (methylene blue 2%, 24 h). The slices were prepared using a microtome. Optical microscope photography was employed to measure the penetration. Results. No statistically significant differences in microleakage were found in the use of bur or laser, nor between adhesive systems. Only statistically significant values were observed comparing enamel with cervical walls (p < 0.001). Conclusion. It can be concluded that the Er:YAG laser is as efficient as diamond bur concerning microleakage values in adhesive restoration procedures, thus constituting an alternative tool for tooth preparation.

Uptake and metabolism of D-glucose in isolated acinar and ductal cells from rat submandibular glands

The present study deals with the possible effects of selected environmental agents upon the uptake and metabolism of d‐glucose in isolated acinar and ductal cells from the rat submandibular salivary gland. In acinar cells, the uptake of d‐[U‐14C]glucose and its non‐metabolised analogue 3‐O‐[14C‐methyl]‐d‐glucose was not affected significantly by phloridzin (0.1 mM) or substitution of extracellular NaCl (115 mM) by an equimolar amount of CsCl, whilst cytochalasin B (20 μM) decreased significantly such an uptake. In ductal cells, both phloridzin and cytochalasin B decreased the uptake of d‐glucose and 3‐O‐methyl‐d‐glucose. Although the intracellular space was comparable in acinar and ductal cells, the catabolism of d‐glucose (2.8 or 8.3 mM) was two to four times higher in ductal cells than in acinar cells. Phloridzin (0.1 mM), ouabain (1.0 mM) and cytochalasin B (20 μM) all impaired d‐glucose catabolism in ductal cells. Such was also the case in ductal cells incubated in the absence of extracellular Ca2+ or in media in which NaCl was substituted by CsCl. It is proposed that the ductal cells in the rat submandibular gland are equipped with several systems mediating the insulin‐sensitive, cytochalasin B‐sensitive and phloridzin‐sensitive transport of d‐glucose across the plasma membrane. Copyright

Comparison between shear forces applied on the overlay-dental tissue interface using different bonding techniques: An in vitro study

Aim: The aim of this study was to compare the adhesion of glass-ceramic overlays to tooth structure, under the effect of shear forces, using different bonding systems. Materials and Methods: Thirty healthy lower third molars were selected and randomly allocated into three groups (n = 10). Group 1: overlays bonded to tooth structure using Panavia V5 with immediate dentin sealing (IDS); Group 2: overlays bonded using Panavia V5 without IDS; and Group 3: overlays bonded using heated composite combined with a bonding agent with IDS. All the restorations were made of glass-ceramic (Suprinity, Vita). The restored teeth were then stored in distilled water for 7 days and at room temperature. Shear forces were applied using a universal testing machine. Load and displacement were recorded at intervals of 0.1 s. A statistical analysis was used to compare the groups. Results: The mean resistance to fractures ± standard deviation obtained for the Groups 1, 2, and 3 was, respectively, 15.7440 ± 2.13, 12.0750 ± 1.41, and 8.33364 ± 2.85 MPa. The analysis of variance was highly significant (P < 0.001) allowing us to reject the null hypothesis of equality between the three groups. Comparisons between pairs also provided significant results. Conclusion: Bonding using Panavia V5 with IDS showed a better resistance to shear forces when compared to other bonding techniques. The application of IDS increased the adhesion.

Influence of the amount of tooth surface preparation on the shear bond strength of zirconia cantilever single-retainer resin-bonded fixed partial denture

PURPOSE Conventional resin-bonded fixed partial dentures (RBFPDs) are usually made with a two-retainer design. Unlike conventional RBFPDs, cantilever resin-bonded fixed partial dentures (Cantilever RBFPDs) are, for their part, made with a single-retainer design. The aim of this study was to compare the effect of tooth surface preparation on the bond strength of zirconia cantilever single-retainer RBFPDs. The objective is to evaluate the shear bond strength of these single-retainer RBFPDs bonded on 3 different amount of tooth surface preparation. MATERIALS AND METHODS Thirty extracted bovine incisors were categorized to 3 groups (n=10), with different amounts of tooth surface preparations. Teeth were restored with single-retainer RBFPDs with different retainer surfaces: large retainer of 32 mm2; medium retainer of 22 mm2; no retainer and only a proximal connecting box of 12 mm2. All RBFPDs were made of zirconia and were bonded using an adhesive system without adhesive capacity. Shear forces were applied to these restorations until debonding. RESULTS Mean shear bond strength values for the groups I, II, and II were 2.39±0.53 MPa, 3.13±0.69 MPa, and 5.40±0.96 MPa, respectively. Statistical analyses were performed using a one-way ANOVA test with Bonferroni post-hoc test, at a significance level of 0.001. Failure modes were observed and showed a 100% adhesive fracture. CONCLUSION It can be concluded that the preparation of large tooth surface preparation might be irrelevant. For zirconia single-retainer RBFPD, only the preparation of a proximal connecting box seems to be a reliable and minimally invasive approach. The differences are statistically significant.

In Vitro Study of Bonding Strength of Zirconia on Dentin Using Different Adhesive Systems

PURPOSE To compare different dental bonding systems that are currently available on the market. MATERIALS AND METHODS A total of 100 extracted, intact third molars were coated in resin, cut, and divided into 10 groups of 10 molars each. Molars were bonded to zirconia blocks with a different bonding system per group. Resistance to shear forces was tested using a universal traction machine. Samples were observed under scanning electron microscopy (SEM) to determine the type of fracture. RESULTS Statistical analyses showed a significant influence of adhesive on the zirconia-dentin assemblys resistance to shear forces. SEM analysis showed mainly adhesive and mixed fractures. CONCLUSION Dentin bonding systems without adhesive capability showed better results than self-etch systems.

Effect of Coloring of Zirconia Framework and Ceramic Veneer on Adhesion of Interfacial Surfaces Determined Using Three-Point Flexural Bonding Strength: An In Vitro Study

PURPOSE The aim of this study was to investigate the effect of coloring on the interfacial surface adhesion between a zirconia framework and ceramic veneer using three-point flexural bonding strength. MATERIALS AND METHODS A total of 40 zirconia bars (Zirlux ST1; DE Healthcare) were cut and divided into two groups of 20 (Groups 1 and 2). The two groups were then further split and divided into four groups of 10 each (Groups 1a, 1b, 2a, and 2b). Groups 1a and 1b tested adhesion of uncolored zirconia and two different shades of ceramic veneer, and Groups 2a and 2b tested adhesion of zirconia colored with two different coloring liquids and one shade of ceramic veneer. RESULTS Some coloring liquids used to color zirconia can significantly affect the bond strength between zirconia and the veneer, whereas ceramic veneer shades do not influence adhesion. CONCLUSION Bonding strength between zirconia framework and ceramic veneer is affected by the coloring of the zirconia.