Biocompatibility of mineral trioxide aggregate and biodentine as root-end filling materials: an in vivo study.

This study evaluated the biocompatibility of mineral trioxide aggregate (MTA) and Biodentine (BD) as root-end filling materials. Six mongrel dogs were divided into two equal groups according to the evaluation period; group A: one month and group B: three months. Three premolars of the same quadrant in each arch were used, summing up 36 teeth (6 teeth/dog). These teeth were randomly subdivided into three subgroups according to the root-end filling material used: MTA, BD and no root-end filling material (control). Endodontic access cavities were performed for induction of periapical pathosis. After the infection period, root canal instrumentation and obturation were accomplished. One day after root canal procedures, root-end surgery was performed. Surgical access was achieved and the root-end was resected approximately 3 mm above the apex. Root-end cavity was prepared ultrasonically and filled with the tested materials. All samples were evaluated by radiography and histopathology (Inflammation and new hard tissue formation). Data were collected and subjected to statistical analysis. In group A, MTA subgroup exhibited significant higher mean inflammatory score than BD subgroup (P < 0.05) while no significant difference was recorded between MTA and BD subgroups in group B (P > 0.05). Regarding mean mineralization score, there was no significant difference between all subgroups in both groups A and B (P > 0.05). Biodentine exhibited favorable biocompatibility in the initial stage of healing than MTA and comparable biomineralization. Clinical relevance: Biodentine could be considered as an acceptable alternative to MTA in peri-radicular surgeries.

An animal study on the effectiveness of platelet-rich plasma as a direct pulp capping agent.

Direct pulp capping (DPC) is a conservative approach for preserving tooth vitality without requiring more invasive procedures by enhancing pulp healing and mineralized tissue barrier formation. We investigated the effectiveness of Platelet Rich Plasma (PRP) vs. Mineral Trioxide Aggregate (MTA) as a DPC agent. Forty-two teeth from three mongrel dogs were divided into two equal groups. After three months, the animals were sacrificed to evaluate teeth radiographically using cone-beam computerized tomography, histopathologically, and real-time PCR for dentin sialophosphoprotein (DSPP), matrix extracellular phosphoglycoprotein (MEPE), and nestin (NES) mRNA expression. Radiographically, hard tissue formation was evident in both groups without significant differences (p = 0.440). Histopathologic findings confirmed the dentin bridge formation in both groups; however, such mineralized tissues were homogenous without cellular inclusions in the PRP group, while was osteodentin type in the MTA group. There was no significant difference in dentin bridge thickness between the PRP-capped and MTA-capped teeth (p = 0.732). The PRP group had significantly higher DSPP, MEPE, and NES mRNA gene expression than the MTA group (p < 0.05). In conclusion, PRP enables mineralized tissue formation following DPC similar to MTA, and could generate better cellular dentinogenic responses and restore dentin with homogenous architecture than MTA, making PRP a promising alternative DPC agent.

In vitro and in vivo evaluation of iRoot BP Plus as a coronal sealing material for regenerative endodontic procedures.

OBJECTIVES: To investigate in vitro effects of a nanoparticle bioceramic material, iRoot BP Plus, on stem cells from apical papilla (SCAP) and in vivo capacity to induce pulp-dentin complex formation. MATERIALS AND METHODS: The sealing ability of iRoot BP Plus was measured via scanning electron microscopy (SEM). SCAP were isolated and treated in vitro by iRoot BP Plus conditioned medium, with mineral trioxide aggregate (MTA) conditioned medium and regular medium used as controls, respectively. Cell proliferation was assessed by BrdU labeling and MTT assay and cell migration was evaluated with wound healing and transwell assays. Osteo/odontogenic potential was evaluated by Alizarin red S staining and qPCR. Pulp-dentin complex formation in vivo was assessed by a tooth slice subcutaneous implantation model. RESULTS: iRoot BP Plus was more tightly bonded with the dentin. There was no difference in SCAP proliferation between iRoot BP Plus and control groups (P > 0.05). iRoot BP Plus had a greater capacity to elevated cell migration (P < 0.05) and osteo/odontogenic marker expression and mineralization nodule formation of SCAP compared with MTA groups (P < 0.05). Furthermore, the new continuous dentine layer and pulp-like tissue was observed in the iRoot BP Plus group in vivo. CONCLUSIONS: iRoot BP Plus showed excellent sealing ability, promoted the migration and osteo/odontogenesis of SCAP and induced pulp-dentin complex formation without affecting the cell proliferation, which indicated iRoot BP Plus was a promising coronal sealing material in REPs. CLINICAL RELEVANCE: The coronal sealing materials play crucial roles for the outcomes of REPs. This study showed that iRoot BP Plus has good coronal sealing and promote pulp-dentin complex formation compared with MTA, providing experimental evidences for the clinical application of iRoot BP Plus as a promising coronal seal material in REPs.

Biocompatibility and Bioactivity of a Dual-Cured Resin-Based Calcium Silicate Cement: In Vitro and in vivo Evaluation.

INTRODUCTION: This study aimed to assess the biocompatibility and bioactivity of a dual-cured resin-based calcium silicate cement in vitro and in vivo. METHODS: For in vitro analyses, standardized samples were prepared using TheraCal LC, TheraCal PT, and ProRoot MTA. The amount of residual monomer released from TheraCal LC and TheraCal PT was assessed using liquid chromatography/mass spectrometry. Calcium ion release from the materials was evaluated using inductively coupled plasma-optical emission spectroscopy. Scanning electron microscopy and energy-dispersive X-ray spectroscopy were used to determine the calcium weight volume in the materials. For in vivo analysis, a rat direct pulp capping model with TheraCal LC, TheraCal PT, and ProRoot MTA groups (n = 16 per group) was used. The rats were euthanized after 7 or 28 days, and histological and immunohistochemical analyses (CD68 and DSPP) were performed. RESULTS: Bisphenol A-glycidyl methacrylate and polyethylene glycol dimethacrylate release from TheraCal PT was lower than that from TheraCal LC (P < .05). Similar results were obtained for calcium-ion release and calcium weight volume, with ProRoot MTA showing the highest values. In the in vivo evaluation, TheraCal PT showed significantly greater hard tissue formation than TheraCal LC (P < .017). TheraCal PT showed lower CD68 expression and greater DSPP expression than TheraCal LC (P < .017). There were no significant differences in the expression of CD68 or DSPP between the TheraCal PT and ProRoot MTA groups. CONCLUSIONS: Within the limitations of this study, the biocompatibility and bioactivity of TheraCal PT could be comparable to those of ProRoot MTA.

Histological evaluation of tissue reaction and new bone formation of different calcium silicate-based cements in rats.

BACKGROUND: The purpose of this study was to evaluate the local reactions and new bone formation of rat subcutaneous and bone tissue to different calcium silicate cements. METHODS: In this study, 80 rats were divided into five groups as control, BIOfactor MTA (BIO), NeoMTA Plus (NEO), MTA Repair Hp (REP), Biodentine (DENT) and then into two subgroups according to sacrification times (7, 30 days; n = 8). Polyethylene tubes filled with appropriate materials (test groups); empty tubes (control group) were implanted into the dorsum of each rat subcutaneously. For intraosseous implantation, materials were placed in the cavities created in tibia of rats. Subcutaneous tissue and tibia samples were stained with haematoxylin-eosin and subjected to histopathological analysis. A score (0-3) was used to grade inflammatory reaction and new bone formation. Data were analysed by Kruskal-Wallis and Mann-Whitney U tests (P < 0.05). RESULTS: Inflammatory reaction observed in subcutaneous and intraosseous tissues for 7 days decreased significantly in all groups over time (P < 0.05). It was determined that there was significant increase in new bone formation in REP, BIO, DENT groups over time (P < 0.05). CONCLUSION: Four contemporary bioceramic materials induced local inflammation and tissues changes shortly after subcutaneous implantation, which were reduced over time. In intraosseous implantation, all materials induced new bone formation over time. REGISTRATION NUMBER: ADJ-03-23-0134. © 2023 Australian Dental Association.

Regenerative potential of a novel aloe vera modified tricalcium silicate cement as a pulp capping material: An animal study.

This study compared the histologic response of a pulp capping material Matreva MTA modified with different concentrations of aloe vera (AV) solutions to Biodentine cement. Ninety dogs' teeth were included and categorized according to the capping material into five groups (18 teeth each); Group I (Biodentine), group II (Matreva MTA), group III (Matreva MTA 10% AV), group IV (Matreva MTA 20% AV) and group V (Matreva MTA 30% AV). The histopathological findings were recorded at 2, 4, and 8 weeks. Matreva MTA and Biodentine groups showed the highest inflammatory cell count compared to the AV-modified Matreva MTA groups at 2- and 4-week intervals (p>0.05). Moreover, the AV-modified Matreva MTA and Biodentine groups showed higher dentin bridge thickness compared to unmodified Matreva MTA at different follow-up periods (p<0.05). AV can significantly enhance the in vivo bioactivity of Matreva MTA, inducing mild inflammation and good dentine bridge formation comparable to Biodentine.

Effect of Fluorohydroxyapatite on Biological and Physical Properties of MTA Angelus.

Objectives: This study aimed to assess the effect of addition of fluorohydroxyapatite (FHI) on biological and physical properties of mineral trioxide aggregate (MTA) Angelus. Materials and Methods: In this in vitro, experimental study, nano-FHI powder was first synthesized, and the morphology and chemical structure of particles were evaluated by scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD). Three groups were evaluated in this study: MTA Angelus, MTA modified with 10% FHA, and MTA modified with 15% FHA. After mixing, the materials were applied to ring molds (10 mm diameter, 1 mm height), and the setting time of the three groups was evaluated according to ISO6876 and ASTMC266-03 with a Gillmore needle. The pH was measured using a pH meter at 24 and 48 hours and 7 days after mixing. The cytotoxicity of the materials was assessed in freshly mixed form and after 1 and 7 days using the methyl thiazolyl tetrazolium (MTT) assay according to ISO10993-5. Data were analyzed by one-way and repeated measures ANOVA and Tukey's test (alpha = 0.05). Results: The addition of FHA to MTA significantly decreased the initial setting time (P < 0.05) and had no significant effect on cell viability (compared with pure MTA Angelus) at 1 and 7 days. However, modified MTA groups in freshly mixed form showed significantly lower cell viability (P < 0.05). The pH remained alkaline at all time points. Conclusion: Addition of 15% FHA to MTA Angelus decreased its setting time with no adverse effect on cell viability (except for fresh form) or pH.

Effect of blood contamination on marginal adaptation of cold ceramic and MTA angelus: a scanning electron microscopic study.

BACKGROUND: This study aimed to assess the effect of blood contamination on marginal adaptation of cold ceramic (CC) and mineral trioxide aggregate (MTA) Angelus using scanning electron microscopy (SEM). METHODS: This in vitro experimental study was conducted on 24 extracted single-rooted human teeth. After cleaning and shaping, the root canals were filled with lateral compaction technique. The apical 3 mm of the roots was cut, and cavities with 3 mm depth were created at the apex. The teeth were randomly assigned to two group (n = 12) for the application of CC and MTA Angelus as retrograde filling materials. CC and MTA Angelus were prepared by mixing the powder with blood, and applied in the cavities. After 24 h, their marginal adaptation to the canal walls was assessed by SEM. Data were statistically analyzed by t-test (alpha = 0.05). RESULTS: The mean marginal gap was 8.98 µm in the CC, and 16.26 µm in the MTA Angelus group; this difference was statistically significant (P < 0.001). CONCLUSIONS: The present in vitro study revealed that following complete blood contamination of powder, CC showed significantly superior marginal adaptation than MTA Angelus as shown by SEM assessment.

Chemomechanical Properties and Biocompatibility of Various Premixed Putty-type Bioactive Ceramic Cements.

INTRODUCTION: This study aimed to evaluate the chemomechanical properties and biocompatibility of recently introduced premixed putty-type bioactive ceramic cements (PPBCs). METHODS: Including ProRoot MTA (PMTA) as a control, BC RRM fast-set putty (BCPT), Well-Root PT (WRPT), One-Fil PT (OFPT), and Endocem MTA premixed (ECPM) were compared to evaluate setting time, radiopacity, pH change, and microhardness. Biocompatibility on human dental pulp cells was compared using CCK-8 assay. Mineralization potential was evaluated using alkaline phosphatase activity, Alizarin Red S (ARS) staining, and quantitative real-time polymerase chain reaction with odontogenic gene marker. For data analysis, 1-way analysis of variance and Tukey's post hoc test were used at the significance level of 95%. RESULTS: Among the PPBCs, BCPT presented the longest (552 ± 27) setting time (minutes) and others showed significantly shorter time than PMTA (334 ± 22) (P < .05). WRPT (6.20 ± 0.54) and OFPT (5.82 ± 0.50) showed significantly higher radiopacity values (mmAl) and others showed similar value compared with PMTA (P > .05). All PPBCs showed high alkaline pH from fresh materials and tended to increase according to time periods from 30 minutes to 12 hours. ECPM showed the highest value of microhardness (81.62 ± 5.90), WRPT showed similar, and others showed lower than PMTA (P < .05). All PPBCs showed biocompatibility in CCK-8 assay. All PPBCs showed similar or better value compared with PMTA in ALP and ARS staining, and ALP and DSPP marker expression (P < .05). CONCLUSIONS: The PPBCs showed clinically acceptable chemomechanical properties and favorable mineralization potential.

Myco-fabricated ZnO nanoparticles ameliorate neurotoxicity in mice model of Alzheimer's disease via acetylcholinesterase inhibition and oxidative stress reduction.

Alzheimer's disease (AD) is one of the primary health problems linked to the decrease of acetylcholine in cholinergic neurons and elevation in oxidative stress. Myco-fabrication of ZnO-NPs revealed excellent biological activities, including anti-inflammatory and acetylcholinesterase inhibitory potentials. This study aims to determine if two distinct doses of myco-fabricated ZnO-NPs have a positive impact on behavioral impairment and several biochemical markers associated with inflammation and oxidative stress in mice that have been treated by aluminum chloride (AlCl3) to induce AD. Sixty male mice were haphazardly separated into equally six groups. Group 1 was injected i.p. with 0.5 ml of deionized water daily during the experiment. Mice in group 2 received AlCl3 (50 mg/kg/day i.p.). Groups 3 and 4 were treated i.p. with 5 and 10 mg/kg/day of ZnO-NPs only, respectively. Groups 5 and 6 were given i.p. 5 and 10 mg/kg/day ZnO-NPs, respectively, add to 50 mg/kg/day AlCl3. Results showed that the AlCl3 caused an increase in the escape latency time and a reduction in the time spent in the target quadrant, indicating a decreased improvement in learning and memory. Moreover, acetylcholinesterase enzyme (AChE) activity and malondialdehyde (MDA), tumor necrosis factor-alpha (TNF-α), and interleukin 1ß (IL-1ß) levels were significantly increased, and the content of glutathione (GSH), activities of superoxide dismutase (SOD), catalase (CAT), alanine aminotransferase (ALT), and aspartate aminotransferase (AST), as well as levels of serotonin and dopamine, were decreased in brain tissues only in AlCl3 treated mice. However, treatment of mice with myco-fabrication of ZnO-NPs at doses of 5 or 10 mg/kg improves learning and memory function through ameliorate all the previous parameters in the AD mice group. The low dose of 5 mg/kg is more effective than a high dose of 10 mg/kg. In accordance with these findings, myco-fabricated ZnO-NPs could enhance memory and exhibit a protective influence against memory loss caused by AlCl3.

Inflammatory response and odontogenic differentiation of inflamed dental pulp treated with different pulp capping materials: An in vivo study.

AIM: Previous studies have evaluated the pulpal responses to calcium silicate cements (CSCs) on normal dental pulp, but investigations on the effects of CSCs on inflamed pulp are limited. This study aimed to test the inflammatory response and odontogenic differentiation of inflamed rat dental pulp after direct pulp capping with CSCs. METHODOLOGY: Wistar rat molars pulps were exposed for 48 h to induce inflammation and then capped with ProRoot MTA (Dentsply), Biodentine (Septodont), RetroMTA (Bio MTA) and Dycal (Dentsply Caulk). The degree of pulpal inflammation and hard tissue formation was evaluated by histological analysis. Immunofluorescence staining for interleukin (IL)-6, osteocalcin (OCN) and runt-related transcription factor 2 (RUNX2) was also performed. RESULTS: After 4 weeks, complete recovery from inflammation was evident in 22%, 37.5%, 10% and none of the ProRoot MTA, Biodentine, RetroMTA and Dycal samples, respectively. Heavy hard tissue deposition as a continuous hard tissue bridge was observed in 77.8%, 75%, 70% and 60% of the ProRoot MTA, Biodentine, RetroMTA and Dycal samples, respectively. IL-6, OCN and RUNX2 were detected in all materials, mainly adjacent to areas of inflammation and reparative dentine formation. At one, two and 4 weeks, significant differences were not observed between the inflammation and hard tissue formation scores of the four material groups (p > .05). CONCLUSIONS: In this study, pulpal inflammation was still present in most specimens at 4 weeks after pulp capping and a significant number of samples showed incomplete and discontinuous dentine bridge formation. The results of this study suggest that initial inflammatory conditions of the pulp may risk the prognosis of teeth treated with CSCs.

Impact of corticosteroid administration on the response of exposed dental pulp to capping with bioactive cements-experimental study on mongrel dogs.

BACKGROUND: Corticosteroids are commonly used as a treatment for a variety of pathological conditions, however, systemic corticosteroid administration has adverse effects including impaired immune response and wound healing. Such complications may affect pulp healing after direct pulp capping. The current study evaluated the influence of corticosteroids on the healing ability of exposed dogs' dental pulps after direct pulp capping (DPC) with bioactive materials. METHODS: Ten healthy male dogs were assigned randomly into two groups, 5 dogs each: group I represent the control group which did not receive any medication, and group II was given corticosteroid for 45 days before DPC and till the dogs were euthanized (n = 75 teeth for each group). Following mechanical exposure, the pulps were randomly capped with either Ca(OH)2, MTA, or Biodentine. The pulpal tissues' reaction to the capping materials was evaluated 65 days postoperatively according to the following parameters: calcific bridge formation, pulpal inflammation, pulp necrosis, and bacterial infiltration. RESULTS: The corticosteroid-treated group revealed no significant difference compared to the control group concerning the pulp healing response (P > 0.05). Both Biodentine and MTA-treated specimens revealed significant differences with Ca(OH)2-treated specimens (P < 0.05) which displayed a superior positive effect of both MTA and Biodentine to Ca(OH)2 regarding all the parameters. CONCLUSIONS: Direct pulp capping technique whenever indicated in subjects treated with corticosteroid immunosuppressive drugs like prednisone performed well in aseptic conditions especially when capped with bioactive materials.

Functional evaluation of mineral trioxide aggregate cement with choline dihydrogen phosphate.

To improve the cytocompatibility of mineral trioxide aggregate (MTA) cement and its ability for reparative dentin formation, the effect of adding choline dihydrogen phosphate (CDHP), which is reported to be biocompatible, to MTA cement was investigated. The L929 cell proliferation showed that the addition of CDHP improved cell viability. The addition of CDHP shortened the setting time of MTA cement, with a significant decrease in consistency above 0.4 g/mL. Diametral tensile strength of the set cement was improved by the addition of 0.4 g/mL CDHP. Solubility was judged to be within the range of clinical application. The spontaneous precipitation of low crystalline hydroxyapatite was examined by immersing the set cement in phosphate buffer saline, and it was found that the ability of the cement with 0.4 g/mL of CDHP was significantly improved compared with that of the cement without CDHP.

Biocompatibility, bioactive potential, porosity, and interface analysis calcium silicate repair cements in a dentin tube model.

OBJECTIVES: This study is to evaluate biocompatibility, bioactive potential, porosity, and dentin/material interface of Bio-C Repair (BIOC-R), MTA Repair HP (MTAHP), and Intermediate Restorative Material (IRM). MATERIALS AND METHODS: Dentin tubes were implanted into subcutaneous of rats for 7, 15, 30, and 60 days. Thickness of capsules, number of inflammatory cells (ICs), interleukin-6 (IL-6), osteocalcin (OCN), and von Kossa were evaluated. Porosity and material/dentin interface voids were also analyzed. Data were submitted to ANOVA and Tukey's tests (p < 0.05). RESULTS: IRM capsules were thicker and contained greater ICs and IL-6-immunopositive cells at 7 and 15 days. BIOC-R capsules exhibited higher thickness and ICs at 7 days and greater IL-6 at 7 and 15 days than MTAHP (p < 0.05). At 30 and 60 days, no significant difference was observed among the groups. OCN-immunopositive cells, von Kossa-positive, and birefringent structures were observed in BIOC-R and MTAHP. MTAHP exhibited higher porosity and interface voids (p < 0.05). CONCLUSIONS: BIOC-R, MTAHP, and IRM are biocompatible. Bioceramics materials demonstrate bioactive potential. MTAHP presented the highest porosity and presence of voids. CLINICAL RELEVANCE: BIOC-R and MTAHP have adequate biological properties. BIOC-R demonstrated lower porosity and presence of voids, which may represent better sealing for its clinical applications.

Mechanism of action of Bioactive Endodontic Materials.

A continuous search for bioactive materials capable of supporting the replacement of damaged pulp tissue, with effective sealing potential and biocompatibility, has represented the attention of studies over the last decades. This study involves a narrative review of the literature developed by searching representative research in PUBMED/MEDLINE and searches in textbooks associated with the mechanism of action of bioactive materials (calcium hydroxide, mineral trioxide aggregate (MTA), and calcium silicate cements). The reflective analysis of the particularities of the chemical elements of these materials, considering the tissue and antibacterial mechanism of action, allows a better understanding of the characteristics and similarities in their tissue responses. Calcium hydroxide paste remains the antibacterial substance of choice as intracanal dressing for the treatment of root canal system infections. Calcium silicate cements, including MTA, show a favorable biological response with the stimulation of mineralized tissue deposition in sealed areas when in contact with connective tissue. This is due to the similarity between the chemical elements, especially ionic dissociation, the potential stimulation of enzymes in tissues, and the contribution towards an alkaline environment due to the pH of these materials. The behavior of bioactive materials, especially MTA and the new calcium silicate cements in the biological sealing activity, has been shown to be effective. Contemporary endodontics has access to bioactive materials with similar properties, which can stimulate a biological seal in lateral and furcation root perforations, root-end fillings and root fillings, pulp capping, pulpotomy, apexification, and regenerative endodontic procedures, in addition to other clinical conditions.

Inflammatory and differentiation cellular response to calcium silicate cements: An in vitro study.

AIM: Inflammatory-regenerative cell interaction is believed to mediate hard tissue formation. This study aimed to investigate the interaction between human inflammatory monocytes with human regenerative fibroblasts after exposure to different calcium silicate materials. METHODOLOGY: Human monocytes were cultured on three materials, polystyrene (PS), mineral trioxide aggregates (MTA) and biodentine (BD), in the presence or absence of lipopolysaccharide (LPS). Half of the monocyte-conditioned media (MoCM) of each group was used to analyse inflammatory cytokine secretion, namely TNF-α, IL-1ß, IL-1RA and IL-6. The remaining MoCM was used to culture recipient fibroblasts, measuring the cell number (proliferation) and levels of alkaline phosphatase (differentiation) and lactic acid dehydrogenase (cytotoxicity). RESULTS: In absence of LPS, MTA was associated with higher secretion of TNF-α and lower secretion of IL-1ß, while BD triggered higher secretions of both cytokines when both materials were compared to control (PS). When LPS was added, higher levels of all analysed cytokines were observed in the PS and BD groups, whereas for the MTA group, only TNF-α and IL-6 were increased. Fibroblasts responded differently to the MoCM from the different groups, revealing significant increases in proliferation and differentiation capacities, particularly when cultured in CM from monocytes exposed to MTA. The morphological evaluation revealed different patterns of fibroblast shape and spread in the different MoCM groups. CONCLUSION: Calcium silicate materials modulate the monocyte inflammatory response, which subsequently induce differential effects on the recipient fibroblasts. MTA appears to promote the secretion of prodifferentiation signals from the monocytes, which are received by fibroblasts, promoting their proliferation and differentiation. The model represents a promising tool to evaluate the interaction of different cells in response to different materials.

Biocompatibility and bioactive potential of an experimental tricalcium silicate-based cement in comparison with Bio-C repair and MTA Repair HP materials.

AIM: To evaluate the tissue reaction of a tricalcium silicate-based repair material associated with 30% calcium tungstate (TCS + CaWO4 ) in comparison to Bio-C Repair (Bio-C; Angelus) and to MTA Repair HP (MTA HP; Angelus). METHODOLOGY: Polyethylene tubes filled with one of the materials or left empty (control group, CG) were implanted into the subcutaneous tissues of rats for 7, 15, 30 and 60 days (n = 32/group). The capsule thickness, number of inflammatory cells, collagen content, interleukin-6 (IL-6), osteocalcin (OCN), von Kossa reaction and analysis under polarized light were evaluated. The data were subjected to generalized linear models for repeated measures, except the OCN. OCN data were submitted to Kruskal-Wallis and Dunn's post hoc test and Friedman followed by Nemenyi's test at significance level of 5%. RESULTS: At all time points, significant differences in the number of inflammatory cells were not observed between TCS + CaWO4 and Bio-C, whereas, at 15, 30 and 60 days, no significant difference was detected between TCS + CaWO4 and MTA HP. At all periods, significant differences were not detected in the number of fibroblasts in TCS + CaWO4 versus MTA HP, and, at 60 days, no significant difference was demonstrated between these groups and CG. Significant differences in the immunoexpression of IL-6 were not detected amongst bioceramic materials at all periods. From 7 to 60 days, significant reduction in the number of inflammatory cells, number of IL-6-immunopositive cells and in the capsule thickness was accompanied by significant increase in the collagen in all groups. OCN-immunolabelled cells, von Kossa-positive structures and amorphous calcite deposits were observed around all materials, whereas, in the CG, these structures were not seen. CONCLUSIONS: These findings indicate that the experimental material (TCS + CaWO4 ) is biocompatible and has a bioactive potential, similar to the MTA HP and Bio-C Repair, and suggest its use as a root repair material.

Biodentine but not MTA induce DSPP expression of dental pulp cells with different severity of LPS-induced inflammation.

OBJECTIVES: To explore the inflammatory and differentiation response in inflamed dental pulp cells (DPCs) induced by lipopolysaccharide (LPS) under different conditions with Biodentine and mineral trioxide aggregate (MTA) treatment. MATERIALS AND METHODS: DPCs were treated with 0.001-1 µg/mL LPS for different periods to induce inflammation. Normal and inflamed DPCs were further treated with 0.14 mg/mL Biodentine or 0.13 mg/mL MTA for different periods. mRNA expression level of IL-6, IL-8 and ALP were analysed by qPCR. DSPP protein expression was detected by western blot. The data were analysed by the Mann-Whitney test, unpaired t test or two-way ANOVA. RESULTS: After treatment for different times and with different concentrations of LPS, different severity of pulp inflammation was revealed by the expressions of IL-6 and IL-8. Higher concentrations of LPS induced higher IL-6 and IL-8 expressions, and these expressions first increased and then decreased (p < 0.0001). At 96 and 192 h, Biodentine significantly suppressed IL-6 expression in both normal and inflamed DPCs (p < 0.05). At 48 and 96 h, Biodentine suppressed ALP expression in both normal and inflamed DPCs (p < 0.05). At 48 and 96 h, Biodentine induced DSPP expressions in both normal and inflamed DPCs (p < 0.05). CONCLUSION: Biodentine enhanced more DSPP differentiation of both normal and inflamed DPCs under different treatment durations than MTA. CLINICAL RELEVANCE: The prognosis of vital pulp therapy may depend on the severity of pulp inflammation which is difficult to be determined in clinical settings. Therefore, Biodentine may enhance odontogenic differentiation in different severity of pulp inflammation imply its clinical indications.

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.

Cell viability and physicochemical effects of different concentrations of bismuth trioxide in a mineral trioxide aggregate cement.

PURPOSE: To evaluate the effect of three concentrations of bismuth trioxide (Bi2O3) on the biological and physicochemical properties of an experimental mineral trioxide aggregate-type (MTA-type) cement at different time points. METHODS: Three experimental groups with white Portland cement containing 15, 20, or 25 wt% of Bi2O3 were assessed. Cellular proliferation in human periodontal ligament fibroblasts was evaluated with an MTT assay. Radiopacity, dimensional stability, pH, and compressive strength were evaluated at different time points. RESULTS: Bismuth trioxide induced cell proliferation in the Bi15 and Bi25 groups in a time-dependent manner; pH was similar in all groups. Compressive strength was associated with time and bismuth concentration. Bi25 had significantly contracted at day 7 and expanded at day 14 (ANOVA P < 0.05, post hoc Tukey test P < 0.05). CONCLUSION: A higher Bi2O3 concentration had a negative effect on the physical properties of the cement at all time points.