Anuradha Prakki

Water distribution in dentin matrices: bound vs. unbound water.

OBJECTIVE This work measured the amount of bound versus unbound water in completely-demineralized dentin. METHODS Dentin beams prepared from extracted human teeth were completely demineralized, rinsed and dried to constant mass. They were rehydrated in 41% relative humidity (RH), while gravimetrically measuring their mass increase until the first plateau was reached at 0.064 (vacuum) or 0.116 gH2O/g dry mass (Drierite). The specimens were then exposed to 60% RH until attaining the second plateau at 0.220 (vacuum) or 0.191 gH2O/g dry mass (Drierite), and subsequently exposed to 99% RH until attaining the third plateau at 0.493 (vacuum) or 0.401 gH2O/g dry mass (Drierite). RESULTS Exposure of the first layer of bound water to 0% RH for 5 min produced a -0.3% loss of bound water; in the second layer of bound water it caused a -3.3% loss of bound water; in the third layer it caused a -6% loss of bound water. Immersion in 100% ethanol or acetone for 5 min produced a 2.8 and 1.9% loss of bound water from the first layer, respectively; it caused a -4 and -7% loss of bound water in the second layer, respectively; and a -17 and -23% loss of bound water in the third layer. Bound water represented 21-25% of total dentin water. Chemical dehydration of water-saturated dentin with ethanol/acetone for 1 min only removed between 25 and 35% of unbound water, respectively. SIGNIFICANCE Attempts to remove bound water by evaporation were not very successful. Chemical dehydration with 100% acetone was more successful than 100% ethanol especially the third layer of bound water. Since unbound water represents between 75 and 79% of total matrix water, the more such water can be removed, the more resin can be infiltrated.

Mechanical properties and characteristics of developmental copolymers incorporating catechin or chlorhexidine

OBJECTIVES This study evaluated catechin and chlorhexidine release from copolymers based on bis-GMA diluted with TEGDMA (R1), propoxylated bis-GMA-CH(3)bis-GMA (R2) or fluorinated bis-GMA-CF(3)bis-GMA (R3). Mechanical properties, degree of conversion, water sorption and solubility were also tested. METHODS Experimental comonomers (R1, R2, R3) were prepared combining bis-GMA with the diluents (70/30 mol%). Each comonomer had either catechin or chlorhexidine incorporated in the following ratios: 0, 0.2, 1.0, 2.0 wt%. The degree of conversion (%DC) of resins was evaluated by FT-IR spectrophotometry (n=3). Flexural strength and elastic modulus were tested on a universal testing machine at a crosshead speed of 0.5mm/min (n=6). Resin disks (7 mm in diameter × 2 mm high) were immersed in deionized water and mass changes recorded daily. Spectral measurements were made to follow any changes in optical densities of deionized water in order to examine catechin and chlorhexidine release rates (n=6). After a 28-day period, water sorption and solubility were calculated using appropriate equations (n=6). Data within the tests were evaluated by one- and three-way ANOVA and Tukey-Kramers test (α=0.05). RESULTS Except for R1 with 2.0% drug none of the groups had changes in degree of conversion and water sorption. Solubility increased with increased drug ratios. Changes in flexural strength and elastic modulus were shown to be variable and material-dependent. All resins presented a ∼24 h burst of drug release. Release rates were drug-ratio dependent and significantly higher for chlorhexidine than for catechin. SIGNIFICANCE Monomer type, %DC, drug ratio and type lead to different interactions between resins and chlorhexidine/catechin drug particles. It should be possible to develop different resins based on clinical needs.

Catechin-incorporated dental copolymers inhibit growth of Streptococcus mutans

Objective: To test the inhibitory growth activity of green tea catechin incorporated into dental resins compared to resins containing the broad-spectrum antimicrobial compound chlorhexidine against Streptococcus mutans in vitro. Material and Methods: The minimum inhibitory concentrations (MICs) of epigallocatechin-gallate (EGCg) and chlorhexidine (CHX) were determined according to the microdilution method. Resin discs (5 mm x 3 mm) were prepared from Bis-GMA/TEGDMA (R1) and Bis-GMA/CH3Bis-GMA (R2) comonomers (n=9) containing: a) no drug, b) EGCg, c) CHX. Two concentrations of each drug (0.5x MIC and 1x MIC) were incorporated into the resin discs. Samples were individually immersed in a bacterial culture and incubated for 24 h at 37º C under constant agitation. Cell viability was assessed by counting the number of colonies on replica agar plates. Statistical analysis was performed using one-way ANOVA, Tukey and Student t-tests (α=0.05). Results: Both resins containing EGCg and CHX showed a significant inhibition of bacterial growth at both concentrations tested (p<0.05). A significantly higher inhibition was observed in response to resins containing CHX at 0.5x MIC and 1x MIC, and EGCg at 1x MIC when compared to EGCg at 0.5x MIC. Also, EGCg at 0.5x MIC in R1 had a significantly higher growth inhibition than in R2. Conclusions: Both EGCg and CHX retained their antibacterial activity when incorporated into the resin matrix. EGCg at 1x MIC in R1 and R2 resins significantly reduced S. mutans survival at a level similar to CHX. The data generated from this study will provide advances in the field of bioactive dental materials with the potential of improving the lifespan of resin-based restorations.

Properties of dental resins submitted to pH catalysed hydrolysis

OBJECTIVES This study evaluated the surface microhardness (SM) and roughness (SR) alterations of dental resins submitted to pH catalysed degradation regimens. METHODS Thirty discs of each TPH Spectrum (Dentsply), Z100 (3M-ESPE), or an unfilled experimental bis-GMA/TEGDMA resin were fabricated, totaling 90 specimens. Each specimen was polymerized for 40 s, finished, polished, and individually stored in deionized water at 37 °C for 7 days. Specimens were randomly assigned to the following pH solutions: 1.0, 6.9 or 13, and for SM or SR evaluations (n = 5). Baseline Knoop-hardness of each specimen was obtained by the arithmetic mean of five random micro-indentations. For SR, mean baseline values were obtained by five random surface tracings (R(a)). Specimens were then soaked in one of the following storage media at 37 °C: (1) 0.1 M, pH 1.0 HCl, (2) 0.1 N, pH 13.0 NaOCl, and (3) deionized water (pH 6.9). Solutions were replaced daily. Repeated SM and SR measurements were performed at the 3-, 7- and 14-day storage time intervals. For each test and resin, data were analysed by two-way ANOVA followed by Tukeys test (α = 0.05). RESULTS There was significant decrease in SM and increase in SR values of composites after storage in alkaline medium. TPH and Z100 presented similar behaviour for SM and SR after immersion in the different media, whereas unfilled resin values showed no significant change. CONCLUSION Hydrolytic degradation of resin composites seems to begin with the silanized inorganic particles and therefore depend on their composition. SIGNIFICANCE To accelerate composite hydrolysis and produce quick in vitro microstructural damage, alkaline medium appears to be more suitable than acidic medium. Contemporary resin composite properties seem to withstand neutral and acidic oral environments tolerably well.

The role of surface sealants in the roughness of composites after a simulated toothbrushing test.

OBJECTIVES To evaluate the influence of two surface sealants (BisCover/Single Bond) and three application techniques (unsealed/conventional/co-polymerization) on the roughness of two composites (Filtek Z250/Z350) after the toothbrushing test. METHODS Seventy-two rectangular specimens (5 mm x10 mm x 3 mm) were fabricated and assigned into 12 groups (n=6). Each sample was subjected to three random roughness readings at baseline, after 100,000 (intermediate), and 200,000 (final) toothbrushing strokes. Roughness (R) at each stage was obtained by the arithmetic mean of the reading of each specimen. Sealant removal was qualitatively examined (optical microscope) and classified into scores (0-3). Data were analyzed by Students paired t-test, two-way ANOVA/Tukeys test, and by Wilcoxon, Kruskal-Wallis and Millers test (alpha=0.05). RESULTS Z250 groups at baseline did not differ statistically from each other. Unsealed Z350 at baseline had lower R values. All the unsealed groups presented gradual decrease in R from baseline to final brushing. From baseline to the intermediate stage, Z250 co-polymerized groups presented a significant reduction in R (score 3). Conventionally sealed groups had no significant changes in R (scores 2-0.8). From baseline to the intermediate stage, the conventionally sealed Z350 Single Bond group had an increase in R (score 1.5). In the final stage, all the conventionally sealed groups presented a reduction in R (scores 0.7-0). Co-polymerized Single Bond groups had a significant reduction in R (scores 2.5-2.7), and co-polymerized BisCover groups an increase in R (scores 2.8-3). CONCLUSIONS At any brushing stage, sealed composites presented superior performance when compared with unsealed composites.

Influence of glutaraldehyde priming on bond strength of an experimental adhesive system applied to wet and dry dentine

OBJECTIVES This study tested the following null hypotheses: (1) there is no difference in resin-dentine bond strength when an experimental glutaraldehyde primer solution is added prior to bonding procedures and (2) there is no difference in resin-dentine bond strength when experimental glutaraldehyde/adhesive system is applied under dry or wet demineralized dentine conditions. METHODS Extracted human maxillary third molars were selected. Flat, mid-coronal dentine was exposed for bonding and four groups were formed. Two groups were designated for the dry and two for the wet dentine technique: DRY: (1) Group GD: acid etching+glutaraldehyde primer (primer A)+HEMA/ethanol primer (primer B)-under dried dentine+unfilled resin; (2) Group D: the same as GD, except for primer A application; WET: (3) Group GW: the same as GD, but primer B was applied under wet dentine condition; (4) Group W: the same as GW, except for primer A application. The bonding resin was light-cured and a resin core was built up on the adhesive layer. Teeth were then prepared for microtensile bond testing to evaluate bond strength. The data obtained were submitted to ANOVA and Tukeys test (alpha=0.05). RESULTS Glutaraldehyde primer application significantly improved resin-dentine bond strength. No significant difference was observed when the same experimental adhesive system was applied under either dry or wet dentine conditions. These results allow the first null hypothesis to be rejected and the second to be accepted. CONCLUSION Glutaraldehyde may affect demineralized dentine properties leading to improved resin bonding to wet and dry substrates.

Indirect pulp treatment in a permanent molar: case reort of 4-year follow-up

This case report describes the Indirect Pulp Treatment (IPT) of deep caries lesion in a permanent molar. A 16-year-old male patient reported discomfort associated with thermal stimulation on the permanent mandibular left first molar. The radiographs revealed a deep distal caries lesion, very close to the pulp, absence of radiolucencies in the periapical region, and absence of periodontal space thickening. Pulp sensitivity was confirmed by thermal pulp vitality tests. Based on the main complaint and the clinical and radiographic examinations, the treatment plan was established to preserve pulp vitality. Clinical procedures consisted of removing the infected dentin and lining the caries-affected dentin with calcium hydroxide paste. The tooth was provisionally sealed for approximately 60 days. After this period, tooth vitality was confirmed, the remaining carious dentin was removed, and the tooth was restored. At 4-year follow-up, no clinical or radiographic pathological findings were found.

Anthocyanin incorporated dental copolymer: bacterial growth inhibition, mechanical properties, and compound release rates and stability by (1)h NMR.

Objective. To evaluate bacterial growth inhibition, mechanical properties, and compound release rate and stability of copolymers incorporated with anthocyanin (ACY; Vaccinium macrocarpon). Methods. Resin samples were prepared (Bis-GMA/TEGDMA at 70/30 mol%) and incorporated with 2 w/w% of either ACY or chlorhexidine (CHX), except for the control group. Samples were individually immersed in a bacterial culture (Streptococcus mutans) for 24 h. Cell viability (n = 3) was assessed by counting the number of colony forming units on replica agar plates. Flexural strength (FS) and elastic modulus (E) were tested on a universal testing machine (n = 8). Compound release and chemical stability were evaluated by UV spectrophotometry and 1H NMR (n = 3). Data were analyzed by one-way ANOVA and Tukeys test (α = 0.05). Results. Both compounds inhibited S. mutans growth, with CHX being most effective (P < 0.05). Control resin had the lowest FS and E values, followed by ACY and CHX, with statistical difference between control and CHX groups for both mechanical properties (P < 0.05). The 24 h compound release rates were ACY: 1.33 μg/mL and CHX: 1.92 μg/mL. 1H NMR spectra suggests that both compounds remained stable after being released in water. Conclusion. The present findings indicate that anthocyanins might be used as a natural antibacterial agent in resin based materials.

Effect of Allium cepa L. on Lipopolysaccharide-Stimulated Osteoclast Precursor Cell Viability, Count, and Morphology Using 4′,6-Diamidino-2-phenylindole-Staining

Allium cepa L. is known to possess numerous pharmacological properties. Our aim was to examine the in vitro effects of Allium cepa L. extract (AcE) on Porphyromonas gingivalis LPS and Escherichia coli LPS-stimulated osteoclast precursor cells to determine cell viability to other future cell-based assays. Osteoclast precursor cells (RAW 264.7) were stimulated by Pg LPS (1 μg/mL) and E. coli LPS (1 μg/mL) in the presence or absence of different concentrations of AcE (10–1000 μg/mL) for 5 days at 37°C/5% CO2. Resazurin reduction and total protein content assays were used to detect cell viability. AcE did not affect cell viability. Resazurin reduction assay showed that AcE, at up to 1000 μg/mL, did not significantly affect cell viability and cellular protein levels. Additionally a caspase 3/7 luminescence assay was used to disclose apoptosis and there was no difference in apoptotic activity between tested groups and control group. Fluorescence images stained by DAPI showed no alteration on the morphology and cell counts of LPS-stimulated osteoclast precursor cells with the use of AcE in all tested concentrations when compared to control. These findings suggest that Allium cepa L. extract could be used for in vitro studies on Porphyromonas gingivalis LPS and Escherichia coli LPS-stimulated osteoclast precursor cells.

Extract of acai-berry inhibits osteoclast differentiation and activity

Osteoclastogenesis is the major cellular event responsible for bone loss and is triggered by inflammation. Acai-berry has proven anti-inflammatory effects. However, there is a lack of evidence for its effects on osteoclastogenesis. Thus, the aim of this study was to determine whether acai-berry extract (ABE) could inhibit osteoclastogenesis and osteoclast activity in vitro. The secretion of cytokines by osteoclasts has been also evaluated. RAW 264.7 cells were stimulated with RANKL (50ng/mL) and treated with various concentrations of ABE (25-100μg/mL) to verify: cell viability (MTT), total protein concentration (BCA), osteoclast differentiation and activity, and cytokine secretion. Cell viability and protein assays showed no toxicity to RAW cells for the tested ABE concentrations (p>0.05). ABE also showed a dose-dependent inhibition of osteoclastogenesis and osteoclast activity evaluated by tartrate-resistant acid phosphatase (TRAP) and hydroxylapatite resorption assay, respectively (p<0.05). ABE decreased the secretion of interleukin (IL)-1α, -6 and tumor necrosis factor alpha while increasing the secretion of IL-3, -4, -13 and interferon gamma when compared to the control group (p<0.05). Results of this study showed that acai-berry extract inhibits osteoclast differentiation and activity possibly due to the modulation of a vast number of cytokines produced by osteoclast precursor cells.