Human Dentine Slabs Research Articles

Eradication of Enterococcus faecalis Biofilms on Human Dentin

Objectives: This work assesses different methods to interfere with Enterococcus faecalis biofilms formed on human dentin slabs.Methods: First, methods are presented that select for small molecule inhibitors of biofilm targets using multi-well polystyrene biofilm plates. Next, we establish methodologies to study and interfere with biofilm formation on a medically relevant model, whereby biofilms are grown on human root dentin slabs.Results: Non-conventional D-amino acid (D-Leucine) can efficiently disperse biofilms formed on dentin slabs without disturbing planktonic growth. Cation chelators interfere with biofilm formation on dentin slabs and polystyrene surfaces, and modestly impact planktonic growth. Strikingly, sodium hypochlorite, the treatment conventionally used to decontaminate infected root canal systems, was extremely toxic to planktonic bacteria, but did not eradicate biofilm cells. Instead, it induced a viable but non-culturable state in biofilm cells when grown on dentin slabs.Conclusion: Sodium hypochlorite may contribute to bacterial persistence. A combination of the methods described here can greatly contribute to the development of biofilm inhibitors and therapies to treat Enterococcus faecalis infections formed in the root canal system.

Scanning electron microscopy analysis of microstructure of the adhesive interface between resin and dentin treated with papain gel.

The aim of this in vitro study was to observe and compare the microstructure of the adhesive interface between resin and dentin treated with Papacarie® using scanning electron microscopy (SEM). Totally, 10 human dentin slabs were randomly distributed into two groups. The control group (n = 5) was subjected to etching with 37% phosphoric acid for 20 s and washed for 30 s, dried with absorbent paper, and the bonding agent was applied along with low viscosity resin. In turn, the experimental group (n = 5) was subjected to the same procedure, but Papacarie® (Fσrmula and Aηγo, Sγo Paulo, Brazil) was added for 30 s prior to etching. Adper™ Single Bond 2 (3M ESPE, Sγo Paulo, Brazil) adhesive was applied to both groups following manufacturer instructions. The specimens were prepared for observation under SEM with ×1.000, ×2.000, ×2.200 and ×5.000 magnification. The micrographs were evaluated with respect to the formation of the hybrid layer, thickness, shape and length of the tags and microtags. In the experimental group there was the formation of more fine hybrid layer and tags with average of similar length to the control group; microtags in less number and without formation of lateral branches. The resin tags presented conical, smooth and uniform characteristics. When Papacarie® was used prior to the application of a bonding agent it could interfere with the formation of the hybrid layer without changing the length of the tag. Moreover, the morphology in the experimental group was found to be more uniform and regular.

Photodynamic antimicrobial therapy of curcumin in biofilms and carious dentine

Photodynamic therapy (PDT) is a technique that involves the activation of photosensitizers by light in the presence of oxygen, resulting in the production of reactive radicals that are capable of inducing cell death. The present study evaluated the susceptibility of Streptococcus mutans and Lactobacillus acidophilus to PDT grown as multi-species in the biofilm phase versus in dentine carious lesions. A brain-heart infusion culture medium supplemented with 1% glucose, 2% sucrose, and 1% young primary culture of L. acidophilus 10(8) CFU/mL and S. mutans 10(8) CFU/mL was used to develop multi-species biofilms and to induce caries on human dentine slabs. Five different concentrations of curcumin (0.75, 1.5, 3.0, 4.0, and 5.0 g/L) were used associated with 5.7 J/cm(2) light emission diode. Four different groups were analyzed L-D- (control group), L-D+ (drug group), L+D- (light group), and L+D+ (PDT group). ANOVA/Tukey's tests were conducted to compare groups. A significant reduction (p <0.05) in cell viability was observed in the biofilm phase following photosensitization with all curcumin concentrations tested. To achieve significant bacterial reduction (p <0.05) in carious dentine, it was necessary to utilize 5.0 g/L of curcumin in association with blue light. No significant reduction was found for L-D+, supporting the absence of the drug's dark toxicity. S. mutans and L. acidophilus were susceptible to curcumin in the presence of blue light. However, due to light penetration and drug diffusion difficulties, these microorganisms within dentine carious lesions were less affected than they were in the biofilm phase.

Antimicrobial effect of chlorhexidine digluconate in dentin: In vitro and in situ study

Aim:The aim of this study was to evaluate a very short-term in vitro and in situ effect of 2% chlorhexidine-digluconate-based (CHX) cavity cleanser on the disinfection of dentin demineralized by cariogenic bacteria.Materials and Methods:Human dentin slabs were randomly allocated and used in 2 distinct phases, in vitro and in situ, for obtaining demineralized dentin. In vitro, the slabs (n=15) were immersed for 5 days in BHI broth inoculated with Streptococcus mutans CTT 3440. In situ, a double-blind design was conducted in one phase of 14 days, during which 20 volunteers wore palatal devices containing two human dental dentin slabs. On 5th day in vitro and 14th day in situ, the slabs were allocated to the two groups: Control group (5 μl of 0.9% NaCl solution) and CHX group (5 μl of 2% chlorhexidine digluconate solution, Cavity Cleanser™ BISCO, Schaumburg, IL, EUA), for 5 minutes. The microbiological analyses were performed immediately before and after the treatments.Results:The log reductions means found for CHX treatment on tested micro organisms were higher when compared to Control group either in vitro or in situ conditions.Conclusions:Our results showed that CHX was effective in reducing the cultivable microbiota in contaminated dentin. Furthermore, although the use of chlorhexidine-digluconate-based cavity disinfectant did not completely eliminate the viable microorganisms, it served as a suitable agent to disinfect tooth preparations.

Effects of Concentrations and Exposure Times of Sodium Hypochlorite on Dentin Deproteination: Attenuated Total Reflection Fourier Transform Infrared Spectroscopy Study

Introduction The purpose of this study was to quantitatively analyze the effects of different concentrations and exposure times of sodium hypochlorite (NaOCl) on dentin deproteination and to determine the suitable concentrations and exposure times to minimize the effects of deproteination on dentin. Methods Intact human dentin slabs were treated with 0.5%, 1%, or 2.25% NaOCl for 1, 5, or 10 minutes. A 0.9% NaCl solution served as the control. To investigate the effect of NaOCl on the surface chemical changes to human dentin, the attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy technique was used to analyze the amide:phosphate ratio and carbonate:phosphate ratio. Results The amide:phosphate ratio decreased significantly after NaOCl treatment as compared with the control group ( p < 0.05). In the 0.5% NaOCl treatment group, the amide:phosphate ratio was significantly higher than that in the 1% and the 2.25% NaOCl treatment groups ( p < 0.05). The different exposure times (1, 5, or 10 minutes) of NaOCl within the same concentration did not influence the amide:phosphate ratio ( p > 0.05). NaOCl treatment did not affect the carbonate:phosphate ratio ( p > 0.05). Conclusions NaOCl, 0.5%, is recommended as the predominant concentration for routine use during root canal therapy to minimize any NaOCl-induced dentin deproteination. It is suggested that a prolonged exposure to low concentrations of NaOCl is less damaging to dentin in attempting to achieve antisepsis during instrumentation of root canal therapy.

In vitro photodynamic antimicrobial chemotherapy in dentine contaminated by cariogenic bacteria

The development of a method to ensure bacterial-free substrates without extensive cavity preparation would be highly useful to dentistry, since there is no currently available effective method for killing residual bacteria in dentinal tissue. This randomized in vitro study determined parameters for using toluidine blue O (TBO) with a light-emitting diode (LED) for dentine caries disinfection and monitored intrapulpal/periodontal temperatures during irradiation. Occlusal human dentine slabs were immersed in Streptococcus mutans culture for demineralization induction. Slabs were allocated to 10 groups (n = 15), which were treated with 0.1 mg ml−1 TBO with 5 min of incubation time or 0.9% NaCl solution for 5, 10 or 15 min, and submitted or not to irradiation for 5, 10 or 15 min (47, 94, and 144 J/cm2). Before and after treatments, dentine samples were analyzed with regard to S. mutans counts. In whole teeth, temperature in pulp and periodontium was measured by thermocouples during irradiation. Kruskal-Wallis/Student-Newman-Keuls, and ANOVA/Tukey test were respectively utilized to compare log reductions and temperature rises between groups. Bacterial reduction was observed when dentine was exposed to both TBO and LED at all irradiation times, as well as to LED alone for 10 and 15 min. Temperature increases lower than 2°C were observed for either pulp or periodontium. Concluding, LED combined with TBO is a safe and effective approach for dentine caries disinfection. Nevertheless, additional studies should be conducted to determine the influence of the irradiation in S. mutans viability in dentinal surface/tubules.

Evaluation of the antimicrobial effect of photodynamic antimicrobial therapy in an in situ model of dentine caries

Photodynamic antimicrobial therapy (PACT) promotes bacterial death as a result of the photosensitization of microbial components. This study evaluated the effect of PACT on dentine caries produced in situ. Over the course of 14 d, 20 volunteers wore intra-oral devices containing human dentine slabs that were treated 10 times daily with a 40% sucrose solution. Afterwards, the antimicrobial effect of toluidine blue O, associated with 47 or 94 J cm(-2) of a light-emitting diode, was evaluated. Before and after the treatments, dentine samples were analysed with regard to the total number of microorganisms, total streptococci, mutans streptococci, and lactobacilli. Significant reductions in the bacterial count were observed for PACT with both energy densities tested, with the following values observed for 47 and 94 J cm(-2) of irradiation: for total streptococci, 3.45 and 5.18; for mutans streptococci, 3.08 and 4.16; for lactobacilli, 3.24 and 4.66; and for total microorganisms, 4.29 and 5.43, respectively. The control, treated with 94 J cm(-2) of irradiation alone, was also effective against all bacteria. To conclude, PACT was effective in killing oral microorganisms present in dentine caries produced in situ and may be a useful technique for eliminating bacteria from dentine carious lesions before restoration.

Effects of plaque control on the patency and occlusion of dentine tubules in situ.

To elucidate the nature of dentine hypersensitivity, the effects of plaque control on the patency and occlusion of dentinal tubules were investigated systematically in situ using human dentine slabs embedded in partial dentures. The dentine slabs were divided into three groups. In group I, the dentine slabs were kept in an oral cavity without plaque control. In group II, plaque was removed mechanically by brushing. Plaque control was carried out chemically using chlorhexidine in group III. After being kept in the oral cavities for 1, 2 and 3 weeks, the slabs were removed from the partial denture, followed by SEM observation to determine the morphological changes of the dentinal tubules. When no attempt was made to remove plaque, the diameter of tubule orifices increased to 390% of the original values within 3 weeks. In contrast, dentinal tubules were found to be occluded, i.e. the tubule orifices became <20% of the original value within 1 week when plaque control efforts were made, using either method of plaque control. We conclude that plaque control plays one of the key roles in the patency versus occlusion of dentinal tubules, and thus in the aetiology and natural reparative process of dentine hypersensitivity.