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Introduction: Osteopontin (OPN), plays an important role in immune system modulation. OPN can activate osteoclasts, thus causing resorption of bone. In addition, it might have a protective function against polymicrobial endodontic infections. Since different isoforms of OPN might have diverse roles, the aim of the present study was to compare gene expression of different isoforms of osteopontin in symptomatic irreversible pulpitis and normal pulps of the human dental pulp. Materials and Methods: Pulps were taken from 20 teeth with symptomatic irreversible pulpitis as the case group and from 20 intact premolars scheduled for extraction as the control group. After RNA extraction and synthesis of complementary DNA (cDNA), quantitative real-time polymerase chain reaction (PCR) was used for the evaluation of gene expression of OPN, OPN2 and OPN3. The Mann-Whitney U, t and Chi-square tests were used to analyze differences between the groups. Results: Mean values of OPN, OPN2 and OPN3 in normal pulps were 0.695±0.295, 0.656±0.298 and 0.816±0.422, respectively. Mean values of OPN, OPN2 and OPN3 in symptomatic irreversible pulpitis were 2.52±1.82, 1.99±0.899 and 1.816±0.954, respectively. Unlike OPN and OPN2, OPN3 exhibited significantly higher expression in normal pulps (P<0.05). Conclusion: The results of the present case-control study showed that some variants of OPN are upregulated during pulpitis and it might be due to their prominent modulatory roles in dental pulps. [ABSTRACT FROM AUTHOR]

The present study aimed to compare the levels of high-mobility group box 1(HMGB1) and soluble triggering receptor expressed on myeloid cells (sTREM1) in the gingival crevicular fluid (GCF). This cross-sectional cohort trial investigated two groups of 22 eligible chronic periodontitis and 22 periodontally healthy individuals (student volunteers) both before and after the periodontal treatment.  GCF was collected from the deepest pockets with clinical attachment loss≥3 mm. Both groups received oral hygiene instructions, and scaling and root planning were performed in the test group. Enzyme-linked immunosorbent assay kit (ELISA) was used to measure the levels of HMGB1 and sTREM1 in GCF samples collected before and 1 month after non-surgical periodontal treatment. The results showed that HMGB1 levels were significantly higher in the chronic periodontitis patients than those of the healthy individuals before treatment (p<0.02) and decreased significantly after periodontal treatment, which reduced gingival inflammation. Furthermore, the levels of sTREM1 marker were significantly higher in periodontitis patients before (p<0.001) and 1 month after treatment than in healthy individuals (p<0.003) although its crevicular levels decreased after periodontal therapy in periodontitis group. The higher levels of sTREM1 and HMGB1 cytokines in GCF of periodontitis patients and the significant decrease after the introduction of the periodontal treatment underlines the importance of HMGB1 and sTREM1 in pathogenesis of periodontitis. [ABSTRACT FROM AUTHOR]

Background: Osteoclastogenesis is coordinated by the interaction of three members of the tumor necrosis factor (TNF) superfamily: Osteoprotegerin (OPG)/receptor activator of nuclear factor kappa B ligand (RANKL)/receptor activator of nuclear factor kappa B (RANK). The aim of this study was to investigate RANKL and OPG levels, and their relative ratio in gingival crevicular fluid (GCF) of patients with chronic and aggressive periodontitis, as well as healthy controls. Materials and Methods: In this analytical study, GCF was obtained from healthy (n = 10), mild chronic periodontitis (n = 18), moderate chronic periodontitis (n = 18), severe chronic periodontitis (n = 20), and generalized aggressive periodontitis (n = 20) subjects. RANKL and OPG concentrations were measured by enzyme-linked immunosorbent assay. Statistical tests used were Kruskal-Wallis test, Mann-Whitney U rank sum test, and Spearman's rank correlation analysis. The level of statistical significance was set at P < 0.05. Results: Mean RANKL concentration showed no statistically significant differences between groups (P = 0.58). There were also no significant differences between mean OPG concentration in the five groups (P = 0.0.56). Moreover, relative RANKL/OPG ratio did not reveal a significant difference between the three study group subjects: healthy, chronic periodontitis (mild, moderate, severe), and aggressive periodontitis (P = 0.41). There was statistically significant correlation between the concentration of sRANKL and Clinical Attachment Level (CAL) in moderate chronic periodontitis patients (R = 0.48, P = 0.04). There was also negative correlation between OPG concentration and CAL in moderate chronic periodontitis patients, although not significant (R = .0.13). Conclusion: RANKL was prominent in periodontitis sites, especially in moderate periodontitis patients, whereas OPG was not detectable in some diseased sites with bleeding on probing, supporting the role of these two molecules in the bone loss developed in this disease. [ABSTRACT FROM AUTHOR]

Objective: Resin cements, regardless of their biocompatibility, have been widely used in restorative dentistry during the recent years. These cements contain hydroxy ethyl methacrylate (HEMA) molecules which are claimed to penetrate into dentinal tubules and may affect dental pulp. Since tooth preparation for metal ceramic restorations involves a large surface of the tooth, cytotoxicity of these cements would be more important in fixed prosthodontic treatments. The purpose of this study was to compare the cytotoxicity of two resin cements (Panavia F2 and Rely X Plus) versus zinc phosphate cement (Harvard) using rat L929-fibroblasts in vitro. Materials and Methods: In this experimental study, ninety hollow glass cylinders (internal diameter 5-mm, height 2-mm) were made and divided into three groups. Each group was filled with one of three experimental cements; Harvard Zinc Phosphate cement, Panavia F2 resin cement and Rely X Plus resin cement. L929-Fibroblast were passaged and subsequently cultured in 6-well plates of 5×105 cells each. The culture medium was RPMI_1640. All samples were incubated in CO2-. Using enzyme-linked immune-sorbent assay (ELISA) and (3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) (MTT) assay, the cytotoxicity of the cements was investigated at 1 hour, 24 hours and one week post exposure. Statistical analyses were performed via two-way ANOVA and honestly significant difference (HSD) Tukey tests. Results: This study revealed significant differences between the three cements at the different time intervals. Harvard cement displayed the greatest cytotoxicity at all three intervals. After 1 hour Panavia F2 showed the next greatest cytotoxicity, but after 24-hours and oneweek intervals Rely X Plus showed the next greatest cytotoxicity. The results further showed that cytotoxicity decreased significantly in the Panavia F2 group with time (p<0.005), cytotoxicity increased significantly in the Rely X Plus group with time (p<0.001), and the Harvard cement group failed to showed no noticeable change in cytotoxicity with time. Conclusion: Although this study has limitations, it provides evidence that Harvard zinc phosphate cement is the most cytotoxic product and Panavia F2 appears to be the least cytotoxic cement over time. [ABSTRACT FROM AUTHOR]

The purpose of this study was to compare substance P (SP) and calcitonin gene-related peptide (CGRP) expression in pulp tissue with clinically diagnosed symptomatic and asymptomatic irreversible pulpitis. Healthy pulps acted as controls. Five normal pulps and 40 with irreversible pulpitis (20 symptomatic and 20 asymptomatic) were obtained from 45 different patients. SP and CGRP expression was determined by competition binding assays using enzyme immunoassay.anova and Mann–Whitney tests were used to ascertain if there were statistically significant differences between the groups. The results showed that neuropeptides were found in all pulp samples. The highest and the lowest expressions for SP and CGRP were found in symptomatic irreversible pulpitis and healthy pulps groups, respectively. The differences between healthy pulps and the groups of pulps having irreversible pulpitis were significant ( P < 0.001). Although Mann–Whitney's post-hoc tests showed statistically significant differences in CGRP expression between two pulpitis groups ( P < 0.05), differences in SP expression between symptomatic and asymptomatic irreversible pulpitis groups were not significant. This study demonstrated that the expression of CGRP and SP is significantly higher in pulps with irreversible pulpitis compared with healthy pulps. [ABSTRACT FROM AUTHOR]

Background: The use of dental pulp stem cells (DPSCs) in clinical applications instead of bone marrow stem cells is a very promising method capable of significantly changing the future of medical treatment. If further studies prove that DPSCs and the cells differentiated from them do not stimulate the immune system, these cells can be used more reliably in treatment of autoimmune diseases., Methods: In this research, we examined the isolated DPSCs and differentiated osteoblasts from them in medium without inflammatory stimulants in terms of TLR3 and TLR4 gene expression and inflammatory cytokines, including TNF-α and IL-8 using qRT-PCR, and measured the concentration of inflammatory cytokines IL-8 and TNF-α produced by these two types of cells through ELISA., Results: The obtained results showed that the expression level of inflammatory cytokines IL-8 and TNF-α in differentiated osteoblasts is significantly different as compared with DPSCs. However, no significant difference was observed in TLR-4 expression between two groups. An increase in TNF-α expression level was found to directly correlate with an increase in the expression of IL-8. The concentration of cytokine TNF-α in osteoblasts was significantly higher than that of IL-8 in DPSCs., Conclusion: In comparison to DPSCs, osteoblast cells first lead to inflammatory responses. These responses reduce overtime. However, DPSCs retain their immunomodulatory properties and do not show inflammatory responses., Competing Interests: None., (AJSC Copyright © 2024.)

Background: Severe early childhood caries (S-ECC) is a multifactorial transmissible infectious disease continuing to affect infants and toddlers worldwide. Saliva plays a modulatory role in the pathogenesis of dental caries., Objectives: The present study aimed to assess the salivary levels of proteinase-3 (PR3) and interleukin-8 (IL-8) as pro-inflammatory cytokines related to the function of neutrophils in association with S-ECC and its treatment., Material and Methods: Fifty children aged 36-60 months were recruited (25 caries-free controls and 25 S-ECC patients). Saliva sampling was performed in all participants. In the S-ECC group, sampling was repeated 6-8 weeks after restorative treatment. The salivary concentrations of PR3 and IL-8 were determined using the enzyme-linked immunosorbent assay (ELISA). The χ2 test, Fisher's exact test, the independent t test, and the paired t test were applied at p < 0.05., Results: The baseline salivary concentrations of PR3 and IL-8 in the S-ECC group were significantly higher than in the caries-free group (p < 0.001). A significant reduction occurred in the levels of these cytokines following restorative treatment in the S-ECC group (p < 0.001), although they were still significantly higher than in the caries-free group (p < 0.05)., Conclusions: The salivary levels of PR3 and IL-8 were significantly affected by the presence of dental caries in children, implying their potential efficiency as non-invasive indicators in the determination of the caries risk and treatment effectiveness.

Background: Dental caries is initiated through mineral dissolution by bacterial acids and collagen degradation by endogenous proteolytic enzymes, mainly collagenolytic matrix metalloproteinases (MMPs)., Objectives: The present research aimed to evaluate the relationship between severe early childhood caries (S-ECC) and salivary MMP-8 and MMP-20 concentrations., Material and Methods: Fifty children aged 36-60 months were assigned to either the caries-free (control) group or the S-ECC group. Standard clinical examinations were performed, and approx. 1 mL of expectorated unstimulated whole saliva was collected from all participants. In the S-ECC group, the sampling was repeated 3 months after restorative treatment. All samples were analyzed for the salivary concentrations of MMP-8 and MMP-20, using the enzyme-linked immunosorbent assay (ELISA). Statistical analysis employed the t test, the Mann-Whitney U test, the χ2 test, Fisher's exact test, and the paired samples t test. The level of significance was set at 0.05., Results: At baseline, the subjects in the S-ECC group presented with significantly elevated levels of MMP-8 as compared to the control group. However, the salivary concentration of MMP-20 did not exhibit a significant difference between the 2 groups. A significant reduction occurred in the levels of MMP-8 and MMP-20 3 months after restorative treatment in the S-ECC group., Conclusions: The salivary levels of MMP-8 and MMP-20 were significantly affected by dental restorative treatment in children. Furthermore, MMP-8 was observed to be a better indicator of the dental caries status than MMP-20.

Introduction: Interleukin-17 is a pro-inflammatory cytokine with a wide range of protective and destructive effects in periodontitis. The role of IL-23 is stabilisation and expansion of Th-17. The aim of this study was to assess whether patients with aggressive and chronic periodontitis exhibit different gingival crevicular fluid (GCF) concentrations of IL-17 and IL-23 compared with clinically healthy subjects., Material and Methods: GCF samples were obtained from 32 patients: 10 with chronic periodontitis (CP), 12 with aggressive periodontitis (AgP), and 10 healthy controls (HC). IL-23 and IL-17 concentrations were measured using enzyme-linked immunosorbent assay (ELISA). Comparison of study groups was performed with ANOVA and Tukey HSD tests. Spearman's correlation coefficient was used to assess correlations between the variables., Results: IL-17 concentration was significantly higher in the healthy group compared to the AgP and CP groups (p < 0.001), but there were no significant differences between the CP and AgP groups. IL-23 levels in the healthy group were significantly higher than that in the AgP group (p < 0.001). Cytokine concentrations did not correlate significantly with probing depths and clinical attachment levels., Conclusions: Gingival crevicular fluid concentrations of IL-17 and IL-23 were significantly higher in the healthy group compared to periodontitis groups.

Objective: Resin cements, regardless of their biocompatibility, have been widely used in restorative dentistry during the recent years. These cements contain hydroxy ethyl methacrylate (HEMA) molecules which are claimed to penetrate into dentinal tubules and may affect dental pulp. Since tooth preparation for metal ceramic restorations involves a large surface of the tooth, cytotoxicity of these cements would be more important in fixed prosthodontic treatments. The purpose of this study was to compare the cytotoxicity of two resin cements (Panavia F2 and Rely X Plus) versus zinc phosphate cement (Harvard) using rat L929-fibroblasts in vitro., Materials and Methods: In this experimental study, ninety hollow glass cylinders (internal diameter 5-mm, height 2-mm) were made and divided into three groups. Each group was filled with one of three experimental cements; Harvard Zinc Phosphate cement, Panavia F2 resin cement and Rely X Plus resin cement. L929- Fibroblast were passaged and subsequently cultured in 6-well plates of 5×10(5) cells each. The culture medium was RPMI&#95; 1640. All samples were incubated in CO2. Using enzyme-linked immune-sorbent assay (ELISA) and (3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) (MTT) assay, the cytotoxicity of the cements was investigated at 1 hour, 24 hours and one week post exposure. Statistical analyses were performed via two-way ANOVA and honestly significant difference (HSD) Tukey tests., Results: This study revealed significant differences between the three cements at the different time intervals. Harvard cement displayed the greatest cytotoxicity at all three intervals. After 1 hour Panavia F2 showed the next greatest cytotoxicity, but after 24-hours and oneweek intervals Rely X Plus showed the next greatest cytotoxicity. The results further showed that cytotoxicity decreased significantly in the Panavia F2 group with time (p<0.005), cytotoxicity increased significantly in the Rely X Plus group with time (p<0.001), and the Harvard cement group failed to showed no noticeable change in cytotoxicity with time., Conclusion: Although this study has limitations, it provides evidence that Harvard zinc phosphate cement is the most cytotoxic product and Panavia F2 appears to be the least cytotoxic cement over time.

Background: Growth factors play a major part in wound healing in many tissues including the periodontium. Transforming growth factor-β1 (TGF-β1) is one of these factors present in the gingival crevicular fluid. In addition, it is considered as one of the most important anti-inflammatory cytokines. Interleukin-1β is a proinflammatory cytokine that presents itself in gingival inflammation and the GCF. Such factors might be of value as prognostic markers of wound healing activity and the therapeutic progress following flap surgery., Objective: The aim of this study was to evaluate the effect of surgical flap on the concentration of IL-1β and TGF-β in the GCF of patients with moderate to severe chronic periodontitis., Methods: The GCF samples were collected, using the Perio-Paper strip at phase 1 (pre-surgery), phase 2 (4th week post surgery) and phase 3 (12th week post surgery) from 20 sites of 10 patients undergoing flap surgery. After the elution, IL-1β and TGF-β concentrations were measured by enzyme-linked immunosorbent assay (ELISA)., Results: The mean TGF-β and IL-1β concentration decreased from phase 1 to phase 3 (p<0.05). There were no significant statistical correlations between IL-1β and TGF-β1 concentrations in the 3 assessment phases. There was a significant statistical correlation between TGF-β1 concentrations and the Plaque Index (PI) in phase 2 (p<0.05). There was a significant statistical correlation (p<0.05) between IL-1β and TGF-β1 concentration and the probing pocket depth (PPD)., Conclusion: The flap surgery has a significant effect on decreasing IL-1β concentration. In the case of TGF-β1, probably the decrease in the concentration after treatment might be due to the removal of the inflammatory stimulants.

Background: The infectious nature of severe early-childhood caries (S-ECC) points to the possible participation of immunologic host responses including neutrophils and their antimicrobial products., Objectives: The aim of this study was to determine the neutrophil apoptosis, α-defensins (HNP1-3) and calprotectin levels in the saliva of preschool children and the association with S-ECC., Methods: Oral examinations were performed on 87 children aged 3-5 years and non stimulated whole saliva samples were collected. Thirty of these subjects were considered S-ECC children, 30 with moderate caries (MC) and 27 were caries free (CF). To detect apoptosis, cell staining was done with Annexin-V-Fluos and propidium iodide, and they were analyzed by fluorescent microscopy. The concentration of α-defensins and calprotectin were assessed using ELISA., Results: There were no statistical differences between groups considering the HNP1-3 or calprotectin salivary levels (p=0.06 and p=0.23, respectively). The HNP1-3 and calprotectin levels were negatively correlated and the correlation was significant in MC group (p=0.03). Lower levels of apoptotic neutrophils were obtained from CF subjects as compared with S-ECC children (p=0.03)., Conclusions: Our findings establish that apoptotic mechanisms could be implicated in the immunity responses associated with S-ECC. We cannot yet determine if the level of salivary α-defensins or calprotectin is predictive of S-ECC.

Background: Receptor activator of nuclear factor kappa B ligand (RANKL) is one of the key cytokines in the induction of osteoclastogenesis both in vitro and in vivo. Several reports indicated the presence of sRANKL in gingival crevicular fluid of patients with periodontal diseases., Objective: To determine the presence of RANKL in peri-implant crevicular fluid samples of implants with peri-implantitis, peri-implant mucositis and healthy controls., Methods: In this study, 40 implants were categorized as clinically healthy, peri-implant mucositis and peri-implantitis according to the clinical and radiographic findings. Filter paper strips were used to collect peri-implant crevicular fluid for 30 seconds in the base of the crevice/pocket. Peri-implant crevicular fluid (PICF) samples were obtained from buccal and lingual aspects of implants. Plaque index, probing depth, gingival index and bleeding on probing were recorded at six sites per implant. Enzyme-linked immunosorbent assay (ELISA) was performed to determine the PICF levels of sRANKL., Results: There were no statistically significant differences in sRANKL concentration between healthy group, peri-implant mucositis and periimplantitis (p=0.12). There were also no statistical correlation between the concentration of sRANKL and probing pocket depth (R=0.051, p=0.65), or any of the other clinical regarding (p>0.05). No differences between the mean sRANKL concentration in the buccal and lingual sites were found (p=0.693)., Conclusion: Our results may suggest that peri-implant crevicular fluid analysis of sRANKL in conjunction with some other osteoclastogenic mediators could be further investigated in further well-designed prospective longitudinal studies on a larger-scale sample size in the evaluation of dental implants.

Background: Early childhood caries (ECC) is a severe type of dental caries affecting infants and pre-school children. Because of the infectious nature of the disease, the immunologic response by the host plays an essential role in its development., Objective: This study investigated the association between the presence of salivary sCD14 and ECC., Methods: This study was carried out on 40 healthy children, of whom 20 were caries-free (CF) and 20 had ECC, within the ages of 36 to 71 months. Unstimulated saliva of the children was collected with disposable needle-less syringe from buccal and labial vestibules. Seven children with ECC received complete treatments. Saliva was collected for a second time after 3 months from this group. The sCD14 levels in salivary samples were analyzed by ELISA method., Results: Mean concentrations of sCD14 in ECC and CF groups were 57.82 and 31.92 ng/ml respectively (p=0.008). After three months, the mean concentration of sCD14 among the treated children decreased to 11.38 ng/ml, which was significantly lower compared to that of ECC children before intervention (p<0.001), and also CF children (p<0.05)., Conclusion: The increased levels of sCD14 can be considered as a marker of inflammation and innate immune response during ECC.

Introduction: Endodontic sealers are in direct contact with periradicular tissues and play a critical role in regeneration and pathogenesis of periradicular diseases. The aim of this study was to compare cytotoxicity of four different types of sealers including AH plus, Sankin, Tubliseal EWT and Apexit as well as their effect on cytokine release of L929 fibroblasts., Materials and Methods: In this experimental study, cells were cultured in Complete Medium Culture (CMC) and then divided into two test groups. In group 1, sealers were added to cell culture wells immediately after mixing. In group 2, sealers were added to cell cultures 3 hours after mixing. Cell viability was evaluated by MTT assay after 4, 24 and 168 hours. The amount of Interleukin-6 (IL-6) released in response to the sealers was also evaluated by ELISA technique on fibroblasts after 24 hour period. Data were analyzed using Kruskal-Wallis and Mann Whitey-U tests., Results: Significant differences were seen in cytotoxicity in both groups (P<0.001). The least cytotoxic sealers were AH Plus and Sankin respectively, whereas Tubliseal EWT showed the greatest cytotoxicity. Production of IL-6 was significantly different among studied groups (P<0.001). The highest IL-6 level was observed in Tubliseal EWT and Sankin groups; which was statistically significant (P<0.001)., Conclusion: AH plus has less cytotoxicity and induces less IL-6 release. Tubliseal EWT has greater cytotoxicity and induces more IL-6 release than other sealers. This should be considered during their routine use in root canal treatments.