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.

Abstract The development of biomaterials capable of driving dental pulp stem cell differentiation into odontoblast-like cells able to secrete reparative dentin is the goal of current conservative dentistry. In the present investigation, a biomembrane (BM) composed of a chitosan/collagen matrix embedded with calcium-aluminate microparticles was tested. The BM was produced by mixing collagen gel with a chitosan solution (2:1), and then adding bioactive calcium-aluminate cement as the mineral phase. An inert material (polystyrene) was used as the negative control. Human dental pulp cells were seeded onto the surface of certain materials, and the cytocompatibility was evaluated by cell proliferation and cell morphology, assessed after 1, 7, 14 and 28 days in culture. The odontoblastic differentiation was evaluated by measuring alkaline phosphatase (ALP) activity, total protein production, gene expression of DMP-1/DSPP and mineralized nodule deposition. The pulp cells were able to attach onto the BM surface and spread, displaying a faster proliferative rate at initial periods than that of the control cells. The BM also acted on the cells to induce more intense ALP activity, protein production at 14 days, and higher gene expression of DSPP and DMP-1 at 28 days, leading to the deposition of about five times more mineralized matrix than the cells in the control group. Therefore, the experimental biomembrane induced the differentiation of pulp cells into odontoblast-like cells featuring a highly secretory phenotype. This innovative bioactive material can drive other protocols for dental pulp exposure treatment by inducing the regeneration of dentin tissue mediated by resident cells.

ResumoIntroduçãoAgentes promotores de ligações cruzadas têm sido investigados como inibidores da atividade enzimática da dentina, o que favoreceria a longevidade das restaurações adesivas.ObjetivoAvaliar o efeito do tratamento da dentina com proantocianidina (PA), em curtos períodos de tempo, na inibição da atividade de MMPs in situ.Material e métodoQuarenta espécimes de dentina (1×1×6 mm) foram obtidos de molares hígidos e divididos em quatro grupos (n=10). Os espécimes foram condicionados com ácido fosfórico por 15 s, seguido de lavagem em água deionizada. A dentina condicionada foi tratada com: água, 5% PA por 5 s, 15 s ou 30 s. A atividade de MMP foi analisada colorimetricamente (SensoLyte®) e os dados de absorbância (412 nm) foram submetidos aos testes de ANOVA e Tukey (α=0,05).ResultadoTodos os períodos de tratamento foram capazes de reduzir a atividade de MMPs, sendo que os melhores resultados foram observados para a dentina tratada com PA por 15 s (63,1% redução) e 30 s (70,2%). O tratamento por 5 s foi capaz de inibir 39,9% das MMPs.ConclusãoA aplicação de PA sobre a dentina condicionada foi capaz de reduzir a atividade de MMPs mesmo em períodos de tempo extremamente curtos, como 5 s. No entanto, melhores resultados foram obtidos com os maiores períodos de tratamento.

AbstractIntroductionCollagen cross-linkers have been investigated as inhibitors of the enzymatic activity of dentin, therefore improving the longevity of adhesive restorations.PurposeTo evaluate the effect of etched dentin treatment with proanthocyanidin (PA) in short periods of time on the inhibition of dentin metalloproteinases (MMP) activity in situ.Material and methodForty dentin specimens (1x1x6mm) were obtained from sound third molars and divided into 4 groups (n=10). The specimens were etched with 37% phosphoric acid for 15s and rinsed in deionized water. Then they were treated with the following solutions: water, 5% PA for 5s, 15s or 30s. The total MMP activity was analyzed by a colorimetric test (SensoLyte®). Absorbance data (412nm) were submitted to ANOVA and Tukey tests (α=0.05).ResultAll treatment periods were able to reduce the total activity of MMPs. The best results were observed for dentine treated with PA for 15s (63.1% reduction) and 30s (70.2%). Treatment for 5s was capable of inhibiting only 39.9% of the total MMP activity.ConclusionApplication of PA on the etched dentin in extremely short periods of time reduced the MMPs activity of dentin, even after 5s. However, the best results were obtained for the longer periods.