Proteolytic activity and degradation of bovine versus human dentin matrices.

OBJECTIVE: Non-human teeth have been commonly used in research as replacements for human teeth, and potential dissimilarities between the dental tissues should be considered when interpreting the outcomes. To compare the proteolytic activity and degradation rate of bovine and human dentin matrices. METHODOLOGY: Dentin beam specimens were obtained from human molars (n=30) and bovine incisors (n=30). The beams were weighed hydrated and after complete dehydration to obtain the mineralized wet and dry masses. Then, the beams were demineralized in 10 wt% phosphoric acid. Next, 15 beams from each substrate were randomly selected and again dehydrated and weighed to obtain the initial demineralized dry mass (DM). Then, the beams were stored in saliva-like buffer solution (SLBS) for 7, 14 and 21 days. SLBS was used to evaluate hydroxyproline (HYP) release after each storage period. The remaining beams of each substrate (n=15) were tested for initial MMP activity using a colorimetric assay and then also stored in SLBS. DM and MMP activity were reassessed after 7, 14 and 21 days of incubation. The data were subjected to two-way ANOVA tests with repeated measures complemented by Bonferroni's tests. Unpaired two-tailed t-tests were also used (p<0.05). RESULTS: Similar water and inorganic fractions were found in human and bovine dentin, while human dentin had a higher protein content. The most intense proteolytic activity and matrix deterioration occurred short after dentin was demineralized. Both substrates exhibited a sharp reduction in MMP activity after seven days of incubation. Although human dentin had higher MMP activity levels, greater HYP release and DM loss after seven days than bovine dentin, after 14 and 21 days, the outcomes were not statistically different. CONCLUSION: Bovine dentin is a suitable substrate for long-term studies involving the degradation of dentin matrices.

Proteolytic activity and degradation of bovine versus human dentin matrices

Abstract Non-human teeth have been commonly used in research as replacements for human teeth, and potential dissimilarities between the dental tissues should be considered when interpreting the outcomes. Objective: To compare the proteolytic activity and degradation rate of bovine and human dentin matrices. Methodology: Dentin beam specimens were obtained from human molars (n=30) and bovine incisors (n=30). The beams were weighed hydrated and after complete dehydration to obtain the mineralized wet and dry masses. Then, the beams were demineralized in 10 wt% phosphoric acid. Next, 15 beams from each substrate were randomly selected and again dehydrated and weighed to obtain the initial demineralized dry mass (DM). Then, the beams were stored in saliva-like buffer solution (SLBS) for 7, 14 and 21 days. SLBS was used to evaluate hydroxyproline (HYP) release after each storage period. The remaining beams of each substrate (n=15) were tested for initial MMP activity using a colorimetric assay and then also stored in SLBS. DM and MMP activity were reassessed after 7, 14 and 21 days of incubation. The data were subjected to two-way ANOVA tests with repeated measures complemented by Bonferroni's tests. Unpaired two-tailed t-tests were also used (p<0.05). Results: Similar water and inorganic fractions were found in human and bovine dentin, while human dentin had a higher protein content. The most intense proteolytic activity and matrix deterioration occurred short after dentin was demineralized. Both substrates exhibited a sharp reduction in MMP activity after seven days of incubation. Although human dentin had higher MMP activity levels, greater HYP release and DM loss after seven days than bovine dentin, after 14 and 21 days, the outcomes were not statistically different. Conclusion: Bovine dentin is a suitable substrate for long-term studies involving the degradation of dentin matrices.

Proteolytic activity, degradation, and dissolution of primary and permanent teeth.

BACKGROUND: Primary and permanent teeth composition may influence dissolution and degradation rates. AIM: To compare the dissolution and degradation of primary and permanent teeth. DESIGN: Enamel and dentin powders were obtained from primary molars and premolars and incubated within different pH buffers. Calcium and inorganic phosphate release was quantified in the buffers by atomic absorption and light spectrophotometry. A colorimetric assay was used to assess the MMP activity of primary dentin (PrD) and permanent dentin (PeD). Collagen degradation was assessed by dry mass loss, change in elastic modulus (E), and ICTP and CTX release. Data were submitted to ANOVA and Tukey's tests (α = 0.05). RESULTS: Similar dissolution was found between PrD and PeD after 256 hours. At pH 4.5, enamel released more minerals than dentin whereas at pH 5.5 the inverse result was observed. MMP activity was similar for both substrates. PrD showed higher dry mass loss after 1 week. In general, greater reduction in E was recorded for PrD. Higher quantities of ICTP and CTX were released from PrD after 1 week. CONCLUSIONS: Primary and permanent teeth presented similar demineralization rates. Collagen degradation, however, was faster and more substantial for PrD.

Inhibitory effect of curcuminoid pretreatments on endogenous dentin proteases.

The aim of this study was to evaluate the effect of curcuminoids on the dentin endogenous protease activity. Demineralized dentin were pretreated with 50 or 100 µM of three different curcuminoids for 60 s and incubated up to 3 months. Untreated beams served as controls. Dry dentin mass was measured after incubation. Aliquots were analyzed for the quantity of ICTP and CTX releases for MMP and cathepsin-K mediated degradation, respectively. The effect of curcuminoids on matrix-bound MMP and soluble rhMMP-9 were measured using an activity assay. Data were subjected to repeated-measures-ANOVA (α=0.05). Gelatinolytic activity was analyzed using zymography. ICTP and CTX release and dry mass loss of curcuminoid-treated groups were significantly lower than the control. Inhibition of rhMMP-9 varied from 29-49% among curcumonoid-treated groups, whereas no inhibition was observed at untreated control (p>0.05). Results were confirmed by zymography. The study showed that the pretreatment of dentin matrices by curcuminoids decreases endogenous protease activity-mediated degradation in dentin.

Can 1% chlorhexidine diacetate and ethanol stabilize resin-dentin bonds?

OBJECTIVES: To examine the effects of the combined use of chlorhexidine and ethanol on the durability of resin-dentin bonds. METHODS: Forty-eight flat dentin surfaces were etched (32% phosphoric acid), rinsed (15 s) and kept wet until bonding procedures. Dentin surfaces were blot-dried with absorbent paper and re-wetted with water (water, control), 1% chlorhexidine diacetate in water (CHD/water), 100% ethanol (ethanol), or 1% chlorhexidine diacetate in ethanol (CHD/ethanol) solutions for 30 s. They were then bonded with All Bond 3 (AB3, Bisco) or Excite (EX, Ivoclar-Vivadent) using a smooth, continuous rubbing application (10 s), followed by 15 s gentle air stream to evaporate solvents. The adhesives were light-cured (20 s) and resin composite build-ups constructed for the microtensile method. Bonded beams were obtained and tested after 24-h, 6-months and 15-months of water storage at 37°C. Storage water was changed every month. Effects of treatment and testing periods were analyzed (ANOVA, Holm-Sidak, p<0.05) for each adhesive. RESULTS: There were no interactions between factors for both etch-and-rinse adhesives. AB3 was significantly affected only by storage (p=0.003). Excite was significantly affected only by treatments (p=0.048). AB3 treated either with ethanol or CHD/ethanol resulted in reduced bond strengths after 15 months. The use of CHD/ethanol resulted in higher bond strengths values for Excite. CONCLUSIONS: Combined use of ethanol/1% chlorhexidine diacetate did not stabilize bond strengths after 15 months.

Effect of reducing acid etching time on bond strength to noncarious and caries-affected primary and permanent dentin.

PURPOSE: The purpose was to evaluate the effect of acid etching time on the bond strength of a simplified etch-and-rinse adhesive system to noncarious and caries-affected dentin of primary and permanent teeth. METHODS: Twenty-four extracted primary and permanent teeth were divided into three groups, according to the acid etching time. Four teeth from each group were exposed to a microbiological caries-inducing protocol. After caries removal, noncarious and caries-affected dentin surfaces were etched with 37 percent phosphoric acid for five, 10, or 15 seconds prior to the application of Prime & Bond NT adhesive. Crowns were restored with resin composite and prepared for microtensile testing. Data were submitted to Kruskal-Wallis and Mann-Whitney tests (α=0.05). RESULTS: Higher bond strengths were obtained for noncarious dentin vs. caries-affected dentin for both primary and permanent teeth. Reducing the acid etching time from 15 to five seconds did not affect the bond strength to caries-affected or noncarious dentin in primary teeth. For permanent teeth, lower bond strength values were observed when the noncarious dentin was etched for five seconds, while no difference was seen between 10 and 15 seconds. CONCLUSIONS: For Prime & Bond NT, the etching of dentin for five seconds could be recommended for primary teeth, while 10 seconds would be the minimum time for permanent teeth.

Effects of a combined application of potassium oxalate gel/adhesive agent on dentin permeability in vitro.

PURPOSE: To test the effects of sequential application of potassium oxalate gel/adhesive agent on in vitro dentin permeability. MATERIALS AND METHODS: Full crown preparations were made in extracted human molars to expose deep coronal dentin. The roots and pulp were removed and the resulting crown segments were connected to a special device (Flodec) to permit the measurement of the permeability of the specimens before and after treatments. Minimum and maximum permeability were recorded after smear layer and phosphoric acid treatment. A new smear layer was created and the permeability measured after the crowns were bonded with Single Bond (3M ESPE), One-Up Bond F (Tokuyama), and AdheSE (Ivoclar Vivadent), either according to manufacturer's instructions or after treating the acid-etched dentin with a 3 wt% potassium oxalate gel. The results were expressed as a percentage of maximum permeability values. Impressions and epoxy resin replicas from the crown segments were produced for SEM examination. RESULTS: None of the adhesives were able to eliminate the fluid flow through dentin. Two-way ANOVA revealed that the application of potassium oxalate prior to the bonding procedures was the most effective technique in reducingthe dentin permeability (p < 0.05), regardless of the adhesive used. SEM micrographs showed that transudation of dentinal fluid could be identified on the surfaces of all replicas. CONCLUSION: The use of potassium oxalate gel was effective in reducing the permeability of bonded dentin.

O paradoxo da evolução dos sistemas adesivos / Evolution of adhesive systems: a paradox

Os adesivos odontológicos são materiais indispensáveis em qualquer procedimento na Odontologia Adesiva da atualidade. Esses vêm passando por significantes modificações de formulação, tornando-os mais simples com relação à técnica de aplicação e mais compatíveis com as características dos substratos dentais. Para a simplificação desses adesivos, houve a necessidade de alterações em sua composição, a qual se destaca pela notável presença de monômeros ácidos, diluentes e água. Com essa formulação, esses agentes tornaram-se mais hidrofílicos e, portanto, mais susceptíveis à absorção de água e conseqüente degradação ao longo do tempo. Neste trabalho, serão enfocados os fenômenos e mecanismos decorrentes da simplificação dos adesivos, objetivando-se alertar o clínico sobre os eventuais inconvenientes que acompanham o avanço tecnológico