Aim To evaluate the tissue response promoted by Bio‐C Pulpo (Bio), MTA Repair HP (MTA‐HP) and White MTA (WMTA) and whether these materials cause liver changes in a rat experimental model. Methodology Polyethylene tubes filled with Bio, MTA‐HP and WMTA, and empty tubes (control group, CG) were implanted into the subcutaneous tissues of rats for 7, 15, 30 and 60 days. Inflammatory reaction score (IRS), capsule thickness, number of inflammatory cells (IC), von Kossa reaction, interleukin‐6 (IL‐6) and alkaline phosphatase (ALP) immunohistochemistry reactions were performed. Combined methods, von Kossa followed by immunohistochemistry for detection of ALP, were performed. At 60 days, the serum glutamic‐oxaloacetic transaminase (GOT) and glutamic‐pyruvic transaminase (GPT) levels were measured and liver fragments were collected for histological analysis; the data were assessed by one‐way ANOVA analysis followed by Sidak's post‐test. The biocompatibility and bioactivity data were subjected to the two‐way ANOVA analysis followed by Tukey post hoc test, except the IRS. The IRS data were subjected to the Kruskal–Wallis ANOVA non‐parametric test followed by Dunn's test (p ≤ .05). Results No significant difference was detected in serum GOT and GPT concentrations and in the number of hepatocytes among the experimental and CG samples. Although Bio‐C Pulpo had the highest IC and IL‐6‐immunolabelled cells (p < 0.0001) at all periods, no significant difference was observed in the IRS among the materials, except at 60 days. In this period, the WMTA had lower IRS. All groups had a significant reduction in the capsule thickness and in the number of IC and IL‐6‐immunolabelled cells over time. Bio‐C Pulpo, MTA‐HP and WMTA specimens had greater immunoexpression of ALP than CG (p < .0001). At all periods, von Kossa‐positive and birefringent structures were observed in the capsules around the materials. ALP‐immunolabelled cells were also seen near von Kossa‐positive structures. Conclusions Bio‐C Pulpo, MTA‐HP and WMTA materials did not cause morphological changes in the liver and no significant alteration in the serum GOT and GPT levels. Moreover, these bioceramic materials were biocompatible and exhibited bioactive potential. However, Bio‐C Pulpo induced greater inflammatory infiltrate than MTA‐HP and WMTA at all periods.

Cytotoxicity, inflammation, biomineralization, and immunoexpression of IL-1β and TNF-α promoted by a new bioceramic cement Aim: To evaluate the cytotoxicity, biocompatibility and mineralization capacity of BIO-C PULPO, and MTA. Methodology: L929 fibroblasts were cultured and MTT assay was used to determine the material cytotoxicity on 6, 24, and 48 h. A total of 30 male rats (Wistar) aged between 4 and 6 months, weighing between 250 and 300 g were used. Polyethylene tubes containing BIO-C PULPO, MTA, and empty tubes were implanted into dorsal connective tissue. After the experimental periods (7, 15, 30, 60, and 90 days) the tubes were histologically analyzed using hematoxylin-eosin (H&E), immunolabeling of IL-1β and TNF-α, and von Kossa staining, or without staining for polarized light analysis. The average number of inflammatory cells was quantified; the mineralization assessment was determined by the area marked in μm2 and semiquantitative immunolabeling analyses of IL-1β and TNF-α were performed. Then, data underwent statistical analysis with a 5% significance level. Results: It was observed that BIO-C PULPO and MTA presented cytocompatibility at 6, 24, and 48 similar or higher than control for all evaluated period. On periods 7 and 15 days, BIO-C PULPO was the material with the highest number of inflammatory cells (p<0.05). On periods 30, 60, and 90 days, BIO-C PULPO and MTA presented similar inflammatory reactions (p>0.05). No statistical differences were found between Control, BIO-C PULPO, and MTA for immunolabeling of IL-1β and TNF-α in the different periods of analysis (p<0.05). Positive von Kossa staining and birefringent structures under polarized light were observed in all analyzed periods in contact with both materials, but larger mineralization area was found with BIO-C PULPO on day 90 (p<0.05). Conclusion: BIO-C PULPO was biocompatible and induced mineralization similar to MTA.

Aim To evaluate the biological properties of experimental sealers based on tricalcium silicate and dicalcium silicate, manipulated with polyethylene glycol (CE‐1) and with the addition of calcium hypochlorite (CE‐2) compared to AH Plus (AHP) and TotalFill BC Sealer (TBC). Methodology The tissue reaction caused by the materials in the subcutaneous tissue of rats was evaluated after implantation of polyethylene tubes filled with the materials at 7, 15, 30 and 60 days. Sections were stained with haematoxylin and eosin (HE) for morphological analysis and to evaluated number of inflammatory cells/mm2 (ICs). Sections were used for immunohistochemical detection of interleukin‐6 (IL‐6) and osteocalcin (OC). The von Kossa method was used to identify calcium precipitation in the capsules. The data were submitted to anova and Tukey’s tests, with 5% significance level. Results At 7 days, CE‐1, CE‐2 and AHP had higher numbers of ICs. AHP presented higher immunolabelling for IL‐6. After 15 days, regarding IL‐6, there was no difference between CE‐2 and the control group. At 30 days, AHP exhibited the highest number of IC (P < 0.05) and CE‐2 and the control group presented the lowest ICs and IL‐6‐positive cells. After 60 days, all materials exhibited decreases in ICs. CE‐2, TBC and the control had the lowest values (P < 0.05). No significant difference was detected between CE‐1 and TBC, and between CE‐2 and control considering the immunoexpression of IL‐6. In this period, AHP had the greatest number of IC and IL‐6 (P < 0.05). In all periods, CE‐1, CE‐2 and TBC sealers had von Kossa‐positive structures and OC‐immunolabelled cells. CE‐2 had higher number of OC‐positive cells than the CE‐1 and TBC sealers (P < 0.05), in all periods. OC immunolabelling was not observed in the capsules of AH Plus and the control groups. Conclusions The experimental sealer and its association with calcium hypochlorite, in addition to TotalFill BC Sealer, were biocompatible and had bioactive potential.

Clavulanic acid (CA) is an important pharmaceutical compound produced by batch fermentation of Streptomyces clavuligerus. Since, CA is chemically unstable, its downstream processing should be studied to develop more efficient and resolute techniques. Herein, the use of aqueous two-phase systems (ATPS) composed of cholinium chloride, [Ch]Cl, was proposed as a novel platform for the recovery and purification of CA. Thus, the stability of CA in presence of different [Ch]Cl concentrations was initially studied, and the high biocompatibility of this salt was demonstrated by the low CA degradation levels. Then, the partitioning of CA using two types of polymeric ATPS has been investigated. Two ATPS formed by the combination of [Ch]Cl and two polyethylene glycol (PEG) polymers, PEG 600 g mol À1 (PEG-600) and polyethylene glycol methyl ether 550 g mol À1 (PEG-500-OMe), were used to assess the influence of the PEG nature, in addition to the concentration of the phase forming agents on the CA partitioning. It has shown that CA is almost equally distributed between the two-phases in equilibrium (0.6 < K CA < 1.6). Nevertheless, the selective extraction of CA for the [Ch]Cl-rich or PEG-rich phase by the proper adjustment of ATPS composition was attained. In the search for higher extraction efficiencies [EE (%)] and partition coefficients (K CA), a second polymeric ATPS platform composed of PEG-600 and sodium polyacrylate 8000 g mol À1 (NaPA-8000), applying [Ch]Cl as adjuvant was tested. The main results suggest the recovery of CA towards the PEG-rich phase (K CA ! 5.6 ± 0.6 and EE ! 85.5± 1.4%). The higher migration levels of CA have mainly resulted from the electronegative repulsion of NaPA-8000 over CA molecules. The ATPS with best performance for the CA extraction were selected for the recovery of CA directly from fermented broth of Streptomyces clavuligerus. In this set of experiments, the highest values of CA recovery yield and purification factor (respectively, 64.91± 1.99% and 22.70 ± 0.87) were attained for the systems PEG-600/NaPA-8000 and PEG-600/[Ch]Cl, respectively.

Objectives: To evaluate the effect of calcium hydroxide (CH) associated with two different vehicles as a capping material for pulp tissue in primary molars, compared with mineral trioxide aggregate (MTA). Methodology: Forty-five primary mandibular molars with dental caries were treated by conventional pulpotomy using one of the following materials: MTA only (MTA group), CH with saline (CH+saline group) and CH with polyethylene glycol (CH+PEG group) (15 teeth/group). Clinical and periapical radiographic examinations of the pulpotomized teeth were performed 3, 6, and 12 months after treatment. Data were tested by chi-squared analysis and a multiple comparison post-test. Results: The MTA group showed both clinical and radiographic treatment success in 14/14 teeth (100%), at all follow-up appointments. By clinical evaluation, no teeth in the CH+saline and CH+PEG groups had signs of mobility, fistula, swelling or inflammation of the surrounding gingival tissue. However, in the CH+saline group, radiographic analysis detected internal resorption in up to 9/15 teeth (67%), and inter-radicular bone resorption and furcation radiolucency in up to 5/15 teeth (36%), from 3 to 12 months of follow-up. In the CH+PEG group, 2/11 teeth (18%) had internal resorption and 1/11 teeth (9%) presented bone resorption and furcation radiolucency at all follow-up appointments. Conclusion: CH with PEG performed better than CH with saline as capping material for pulpotomy of primary teeth. However, both combinations yielded clinical and radiographic results inferior to those of MTA alone.

The aim of this preliminary study was to compare the effects of different energy densities from red and infrared low-level laser (LLL) on viability and proliferation of stem cells from human exfoliated deciduous teeth (SHED). SHED were irradiated with red laser (R) or infrared laser (IR) set with the following dosimetry: 1.2 J/cm (0.05 J), 2.5 J/cm (0.1 J), 5.0 J/cm (0.2 J), and 7.5 J/cm (0.3 J). Positive (C+) and negative (C-) control groups comprised non-irradiated cells. Data were analyzed by two-way ANOVA followed by Tukey's test (P < 0.05). At 24- and 48-h period, group R5.0 showed significantly higher cell viability rates than R1.2 and R2.5. At 48 h, R2.5 also revealed lower proliferation than R5.0. Comparing to the C+ group, R2.5 exhibited lower viability at 72 h, and proliferation at 24 and 48 h. Groups R1.2, IR1.2, and IR5.0 were less viable at 24 h, while R1.2, IR2.5, and IR5.0 revealed lower proliferative capacity at 48 h. Overall, our results showed that LLL can favor viability and proliferation of SHED, especially when cells receive red laser irradiation at 5.0 J/cm. Therefore, according to this preliminary investigation, 5 J/cm applied by red LLL induced high rates of cell viability and proliferation, while the same irradiation dose using infrared laser led to negative effects. LLL irradiation with 1.2 and 2.5 J/cm was deleterious to metabolic activity and proliferation of SHED regardless of the type of laser. Further studies are necessary to gain in-depth knowledge about the effects of different wavelengths of LLL on SHED viability and proliferation.

Our purpose was to evaluate the biocompatibility and hepatotoxicity of a new bioceramic intracanal medicament, Bio-C Temp (BIO). The biological properties of BIO were compared with calcium hydroxide-based intracanal medicament (Calen; CAL), used as gold pattern. Polyethylene tubes filled with BIO or CAL, and empty tubes (control group, CG) were implanted into subcutaneous tissue of rats. After 7, 15, 30 and 60 days, the samples were embedded in paraffin for morphological, quantitative and immunohistochemistry analyses. At 7 and 60 days, blood samples were collected for analysis of serum glutamic-oxaloacetic transaminase (GOT) and glutamic-pyruvic transaminase (GPT) levels. The data were submitted to two-way ANOVA and Tukey’s test (p ≤ 0.05). No significant difference was detected in serum GOT and GPT levels among BIO, CAL and CG specimens. In all periods, BIO specimens exhibited lower number of inflammatory cells and immunoexpression of IL-6, a pro-inflammatory cytokine, than CAL specimens. The reduction of these parameters was accompanied by significant increase in the collagen content and in the immunoexpression of IL-10, a cytokine involved in the tissue repair, over time. Our findings indicate that Bio-C Temp is biocompatible and had no hepatotoxicity effect.

Purpose To compare, both qualitatively and quantitatively, the inflammatory cells, vascular density and IL-6 immunolabeled cells present in the pulp after pulpotomy with white MTA versus 15.5% ferric sulfate (FS). Methodology Forty-eight mandibular first molars from 24 Wistar rats were divided into MTA or FS groups and subdivided according to the period after pulpotomy procedure (24, 48 and 72 hours). Four teeth (sound and untreated) were used as controls. Histological sections were obtained and assessed through the descriptive analysis of morphological aspects of pulp tissue and the quantification of inflammatory cells, vascular density and interleukin-6 (IL-6) expression. Data were statistically analyzed (p<0.05). Results The number of inflammatory cells was similar in both groups, being predominantly localized at the cervical radicular third. In the MTA group, increased inflammation was observed at 48 hours. Vascular density was similar in both groups and over time, being predominant in the medium radicular third. No correlation was found between the number of inflammatory cells and the vascular density. Pulp tissue was more organized in MTA-treated teeth. In both groups, a weak to moderate IL-6 expression was detected in odontoblasts and inflammatory cells. Comparing both groups, there was a greater IL-6 expression in the cervical radicular third of teeth treated with MTA at 24 hours and in the medium and apical thirds at 72 hours, while in the FS group a greater IL-6 expression was found in the apical third at 24 hours. Conclusion The MTA group presented better histological features and greater IL-6 expression than the FS group. However, no difference was observed between the groups regarding the inflammatory status and vascularization, suggesting the usefulness of FS as a low-cost alternative to MTA.