Comparison of neutrophil apoptosis alpha-defensins and calprotectin in children with and without severe early childhood caries

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. The aim of this study was to determine the neutrophil apoptosis, alpha-defensins [HNP1-3] and calprotectin levels in the saliva of preschool children and the association with S-ECC. 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 alpha-defensins and calprotectin were assessed using ELISA. 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]. 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 alpha-defensins or calprotectin is predictive of S-ECC

Effect of surgical flap on IL-1 beta and TGF-beta concentrations in the gingival crevicular fluid of patients with moderate to severe chronic periodontitis

Growth factors play a major part in wound healing in many tissues including the periodontium. Transforming growth factor-beta1 [TGF-beta1] 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-1 beta is a proinflammatory cytokine that presents itself in gingival inflammation and the GCF. Such factors might be of value as prognostic makers of wound healing activity and the therapeutic progress following flap surgery. The aim of this study was to evaluate the effect of surgical flap on the concentration of IL-1beta and TGF-beta in the GCF of patients with moderate to severe chronic periodontitis. 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 enzyme-linked immunosorbent assay [ELISA]. The mean TGF-beta and IL-1beta statistical correlations between Il-1beta and TGR-beta1 concentrations in the 3 assessment phases. There was a significant statistical correlation between TGF-beta1 concentrations and the Plaque Index [PI] in phase 2 [p<0.05]. There was a significant statistical correlation [p<0.05] between IL-1beta and TGF-beta 1 concentration and the probing pocket depth [PPD]. The flap surgery has a significant effect on decreasing IL-1beta concentration. In the case of TGF-beta1, probably the decrease in the concentration after treatment might be due to the removal of the inflammatory stimulants

Cytotoxicity comparison of harvard zinc phosphate cement versus panavia F2 and rely X plus resin cements on rat L929-fibroblasts

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. 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 5x10[5] cells each. The culture medium was RPMI_ 1640. All samples were incubated in CO[2]. 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. 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. 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

Association of salivary sCD14 concentration levels with early childhood caries

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. This study investigated the association between the presence of salivary sCD14 and ECC. 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. 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]. The increased levels of sCD14 can be considered as a marker of inflammation and innate immune response during ECC

Receptor activator of nuclear factor kappa B ligand [RANKL] levels in peri-implant crevicular fluid

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. To determine the presence of RANKL in peri-implant crevicular fluid samples of implants with peri-implantitis, peri-implant mucositis and healthy controls. 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. 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]. 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

Evaluation of cytotoxicity of different root canal sealers and their effect on cytokine production

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. 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. 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]. 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

Assessment of anti-streptokinase antibody in patients with heart diseases and normal subjects

Background: Streptokinase, which is injected intravenously with a standard dose of 1.5 MIU, is the most widely used thrombolytic agent around the world. What is so important about this bioproduct is the level of anti-streptokinase [anti-sk] antibody in the population, which is directly correlated to the incidence of streptococcal infections in that population

Objectives: Since Iran is an endemic area for streptococcal infections, this study was conducted to assess the anti-sk level in an Iranian population

Materials and Methods: 97 males and 47 females referred to Modarress Hospital of Tehran for coronary angiography and cardiac catheterization were included. 10 ml of venous blood was taken before angiographies from each patient. According to the angiography reports, the patients were divided into three groups: Coronary Artery Diseases [CAD, n=95], Rheumatic Heart Disease [RHD, n=19] and normal coronaries [n=30]. The anti-sk antibody level was assessed in the serum samples of all patients using Enzyme Linked Immunosorbant Assay

Results: In 23.2% of patients with CAD, 40% of normal coronaries and 73.7% of patients with RHD, the serum samples contained more than 2 arbitrary units [AU] of anti-SK antibody which regarded as high levels. There was no significant difference between the anti-sk level of patients with CAD and normal coronaries [2.03 +/- 3.02 AUs vs. 2.52 +/- 2.23 AU], but the level of antibody in RHD group [8.16 +/- 10.1 AU] was significantly higher than other groups [p<0.05]. No significant correlation was observed between antibody levels and the age or gender of patients

Conclusion: We concluded that the level of anti-sk antibody is high in Iranian population as compared to other endemic areas for streptococcal infections. Also we found no relation between the level of antibody and sex and age of patients. This study accentuated the necessity of assessment of drug efficacy in endemic areas for streptococcal infections especially in those patients with valvular heart disease