Bioactive potential of Bio-C Temp demonstrated by systemic mineralization markers and immunoexpression of bone proteins in the rat connective tissue.

Intracanal medications are used in endodontic treatment due to their antibacterial activity and ability to induce the periapical repair. Among the intracanal medications, the Calen (CAL; SS. White, Brazil) is a calcium hydroxide-based medication that provides an alkaline pH and releases calcium, exerting an antimicrobial activity. Bio-C Temp (BIO; Angelus, Brazil), a ready-to-use bioceramic intracanal medication, was designed to stimulate the mineralized tissues formation. Here, we investigated the bioactive potential of BIO in comparison to the CAL in the rat subcutaneous. Polyethylene tubes filled with medications, and empty tubes (control group, CG) were implanted in the subcutaneous tissue of rats. After 7, 15, 30 and 60 days, the blood was collected for calcium (Ca+2) and alkaline phosphatase (ALP) measurement, and the capsules around the implants were processed for morphological analyses. The data were submitted to two-way ANOVA and Tukey test (p < 0.05). At 7, 15 and 30 days, the ALP level was grater in BIO and CAL than in CG (p < 0.0001). At 7 and 15 days, greater Ca+2 level was seen in the serum of CAL samples. From 7 to 60 days, an increase in the number of fibroblasts, osteocalcin- and osteopontin-immunolabelled cells was observed in BIO and CAL groups (p < 0.0001). In all periods, BIO and CAL specimens showed von Kossa-positive structures. Moreover, ultrastructural analysis revealed globules of mineralization in the capsules around the BIO and CAL specimens. Thus Bio-C Temp caused an increase in the ALP, osteocalcin and osteopontin, which may have allowed the formation of calcite, suggesting bioactive potential.

Participation of fibroblast growth factor-1 and interleukin-10 in connective tissue repair following subcutaneous implantation of bioceramic materials in rats.

AIM: To evaluate whether the bioceramic materials Bio-C Pulpo (Bio-C, Angelus) and mineral trioxide aggregate (MTA) Repair HP (MTA-HP, Angelus) induce fibroblast proliferation and release of interleukin-10 (IL-10), an anti-inflammatory cytokine, stimulating connective tissue remodelling. The tissue response of Bio-C and MTA-HP was compared with the White MTA (WMTA; Angelus) since studies have demonstrated that WMTA induces tissue repair. METHODOLOGY: Bio-C, MTA-HP and WMTA were inserted into polyethylene tubes and implanted in the subcutaneous tissue of Holtzman rats for 7, 15, 30 and 60 days. As a control group (CG), empty tubes were implanted subcutaneously. The number of fibroblasts (FB), Ki-67-, fibroblast growth factor-1- (FGF-1) and IL-10-immunolabelled cells and collagen content in the capsules was obtained. The data were subjected to two-way anova followed by Tukey's test (p ≤ .05). RESULTS: At 7 days, significant differences in the number of FB were not detected amongst Bio-C, MTA-HP and WMTA groups (p Ëƒ .05). The capsules of all groups exhibited a significant increase in the number of FB and content of collagen over time. From 7 to 60 days, a significant reduction in the number of FGF-1- and Ki-67-immunolabelled cells was seen in the capsules of all specimens. In all periods, no significant difference in the number of FGF-1-immunolabelled cells was detected between Bio-C and CG specimens. At 60 days, significant differences in the immunoexpression of FGF-1 were not observed amongst the groups. At 7 and 15 days, the highest immunoexpression for Ki-67 was present in Bio-C specimens whilst, after 30 and 60 days, no significant difference was observed amongst the bioceramic materials. At 7 days, few IL-10 immunolabelled cells were present in the capsules of all specimens whereas, at 60 days, a significant increase in the IL-10-immunostaining was present in all groups. At 60 days, the Bio-C, MTA-HP and WMTA groups showed a greater number of IL-10-immunolabelled cells than in the CG specimens (p < .0001). CONCLUSIONS: Bio-C, MTA-HP and WMTA stimulate fibroblast proliferation, leading to the formation of collagen-rich capsules. FGF-1 and IL-10 may mediate the remodelling of capsules around Bio-C, MTA-HP and WMTA bioceramic materials.

Hepatic enzymes and immunoinflammatory response to Bio-C Temp bioceramic intracanal medication implanted into the subcutaneous tissue of rats.

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.

Comparison of Bio-C Pulpo and MTA Repair HP with White MTA: effect on liver parameters and evaluation of biocompatibility and bioactivity in rats.

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.

Biocompatibility and bioactive potential of the NeoMTA Plus endodontic bioceramic-based sealer.

OBJECTIVES: This study evaluated the biocompatibility and bioactive potential of NeoMTA Plus mixed as a root canal sealer in comparison with MTA Fillapex. MATERIALS AND METHODS: Polyethylene tubes filled with NeoMTA Plus (n = 20), MTA Fillapex (n = 20), or nothing (control group, CG; n = 20) were inserted into the connective tissue in the dorsal subcutaneous layer of rats. After 7, 15, 30 and 60 days, the specimens were processed for paraffin embedding. The capsule thickness, collagen content, and number of inflammatory cells (ICs) and interleukin-6 (IL-6) immunolabeled cells were measured. von Kossa-positive structures were evaluated and unstained sections were analyzed under polarized light. Two-way analysis of variance was performed, followed by the post hoc Tukey test (p ≤ 0.05). RESULTS: At 7 days, the capsules around NeoMTA Plus and MTA Fillapex had more ICs and IL-6-immunostained cells than the CG. However, at 60 days, there was no significant difference in the IC number between NeoMTA Plus and the CG (p = 0.1137) or the MTA Fillapex group (p = 0.4062), although a greater number of IL-6-immunostained cells was observed in the MTA Fillapex group (p = 0.0353). From 7 to 60 days, the capsule thickness of the NeoMTA Plus and MTA Fillapex specimens significantly decreased, concomitantly with an increase in the collagen content. The capsules around root canal sealers showed positivity to the von Kossa stain and birefringent structures. CONCLUSIONS: The NeoMTA Plus root canal sealer is biocompatible and exhibits bioactive potential.

Biocompatibility and Bioactive Potential of New Calcium Silicate-based Endodontic Sealers: Bio-C Sealer and Sealer Plus BC.

INTRODUCTION: Bio-C Sealer (BC; Angelus, Londrina, PR, Brazil) and Sealer Plus BC (SPBC; MK Life, Porto Alegre, Brazil) are new ready-to-use bioceramic endodontic sealers. The aim of this study was to evaluate the biocompatibility and bioactive potential of BC and SPBC sealers in comparison with AH Plus (AHP; Dentsply DeTrey Gmbh, Konstanz, Germany) in subcutaneous tissue of rats. METHODS: Polyethylene tubes filled with materials and empty tubes, serving as the control group, were implanted in the subcutaneous tissues of rats. After 7, 15, 30, and 60 days, the tubes with connective tissue were removed, and inflammatory cells (ICs)/mm2 and immunolabeled cells for interleukin (IL)-6 were evaluated. Osteocalcin and von Kossa analysis were also performed. Data were submitted to analysis of variance and Tukey tests, with a significance level of 5%. RESULTS: At 7 days, SPBC showed lower ICs than BC (P < .05). AHP exhibited greater immunolabeled cells for IL-6 (P < .05). After 15 days, BC showed lower ICs and IL-6 compared with other materials. At 30 days, SPBC and AHP showed higher values for ICs (P < .05). After 60 days, calcium silicate sealers did not show statistical difference (P > .05) for ICs and IL-6, with values lower than AHP (P < .05). The materials showed positive structures to von Kossa staining. BC exhibited osteocalcin labeling in all periods. SPBC showed osteocalcin labeling from 15-60 days. AHP and the control group did not exhibit osteocalcin labeling. CONCLUSIONS: BC and SPBC sealers are biocompatible and present bioactive potential.

Immunoinflammatory response and bioactive potential of GuttaFlow bioseal and MTA Fillapex in the rat subcutaneous tissue.

To evaluate the effect of GuttaFlow bioseal (GFB) and MTA Fillapex (MTAF) in comparison with Endofill (EF) in the subcutaneous tissue. Polyethylene tubes with GFB, MTAF, EF or empty tubes (control group; CG) were implanted into subcutaneous of rats. After 7, 15, 30 and 60 days, the capsule thickness, inflammatory reaction, interleukin-6 (IL-6), vascular endothelial growth factor (VEGF), caspase-3, TUNEL-positive cells, von Kossa and ultrastructural features were evaluated. The data were statistically analyzed (p ≤ 0.05). At all periods, the number of IL-6- and VEGF-immunolabelled cells, and capsule thickness were lower in GFB than MTAF, which was lower than EF (p < 0.0001). At 60 days, the number of inflammatory cells was similar in GFB and MTAF (p = 0.58). Significant differences in the number of TUNEL- and caspase-3-positive cells were not observed among GFB, MTAF and CG whereas the highest values were found in EF specimens. The EF specimens exhibited several cells with condensed chromatin, typical of apoptosis. von Kossa-positive and birefringent structures were only observed in GFB and MTAF, suggesting the presence of calcite crystals. Taken together, these results show that cellular and structural damage induced by GFB and MTAF sealers were recovery over time. Moreover, these sealers express bioactive potential in subcutaneous tissue.

Physical Properties, Antimicrobial Activity and In Vivo Tissue Response to Apexit Plus.

We investigated the physical properties, antimicrobial activity, and tissue reaction to Apexit Plus in comparison to Sealapex. Flow, radiopacity, setting time, and solubility were evaluated in each material. The antimicrobial activity against Enterococcus faecalis was performed. Polyethylene tubes containing Apexit Plus or Sealapex, and without material (control group) were implanted into the subcutaneous tissue of rats. At 7, 15, 30, and 60 days of implantation, the specimens were paraffin-embedded and the number of inflammatory cells (ICs) and the amount of birefringent collagen (BC) were quantified. The von Kossa reaction followed by immunohistochemistry for detection of alkaline phosphatase (ALP) was also performed. Statistical analysis was performed with ANOVA and Tukey test (p ≤ 0.05). The flow value of Apexit Plus was greater than Sealapex, whereas the radiopacity (3.44 mm Al) was lower than Sealapex (6.82 mm Al). Apexit Plus showed lower solubility and shorter initial and final setting (p 0.0001), whereas the antimicrobial activity was significantly greater than Sealapex. Although the number of ICs was higher in Apexit Plus (p = 0.0009) at 7 days, no significant difference was detected between Apexit Plus and Sealapex at 15, 30, and 60 days. All groups showed higher values for BC in the capsules over time. ALP-immunolabelled cells were observed, mainly around von Kossa-positive structures, either in the capsules of Apexit Plus or Sealapex. Therefore, our results revealed that Apexit Plus exhibited a greater effectiveness against Enterococcus faecalis and better physical properties than Sealapex, except for the radiopacity. In vivo findings indicate that Apexit Plus is biocompatible and presents potential bioactivity in the subcutaneous tissue.

Biodentine and MTA modulate immunoinflammatory response favoring bone formation in sealing of furcation perforations in rat molars.

OBJECTIVES: Evaluate the tissue reaction of periodontium subjacent to furcation perforations in rat molars sealed with Biodentine or mineral trioxide aggregate (MTA). MATERIALS AND METHODS: The pulp chamber floor of right upper first molars of 60 rats was perforated and filled with Biodentine, MTA, or cotton pellet (sham); the left first molars were used as control. After 7, 15, 30, and 60 days, maxillary fragments were processed for paraffin-embedding. The periodontal space (PS), volume density of inflammatory cells (VvIC) and fibroblasts (VvFb), number of osteoclasts, and collagen content were obtained. Interleukin-6 (IL-6) and osterix (osteoblast marker) were detected by immunohistochemistry. The data were submitted to ANOVA and Tukey's test (p ≤ 0.05). RESULTS: At 7 days, high values in VvIC, IL-6-immunolabeled cells, and osteoclasts were accompanied by reduced collagen content in enlarged PS of experimental groups. At all periods, VvIC, number of osteoclasts and IL-6, and PS were higher in sham than in Biodentine and MTA (p < 0.0001). From 7 to 60 days, significant reduction in VvIC, IL-6 immunoexpression, and osteoclasts was accompanied by significant increase in VvFb, osteoblasts, and collagen in Biodentine and MTA groups. At 60 days, significant differences in VvIC, PS, IL-6, osteoclasts, and osteoblasts were not found between Biodentine and MTA. Significant differences in the osteoclast number were not observed among Biodentine, MTA, and control groups while osteoblasts number was higher in Biodentine and MTA groups. CONCLUSIONS: Despite the initial inflammatory reaction and bone resorption, the sealing of furcation perforations with Biodentine and MTA favors the repair of periodontal tissues. CLINICAL RELEVANCE: Biodentine and MTA exhibit potential as repair material in the treatment of furcation perforations.