Influence of resveratrol application with pulp-capping materials on the genetic expression levels of stem cells.

AIM: To evaluate in a laboratory setting the response of human mesenchymal stem cells (MSCs) to pulp-capping materials with and without resveratrol (RSV). METHODOLOGY: Five materials, Calcimol LC, Life, TheraCal LC, ProRoot MTA and Biodentine, were prepared according to the manufacturers' instructions. Human MSCs were then exposed to these materials, with and without RSV, for 24 h (n = 8). Cell viability was evaluated using the MTT assay, and total cell death was quantified by annexin V-FITC staining with flow cytometry. The expression levels of the IL-8, IL-10, HBD-2 and BCL-2 genes were investigated using real-time polymerase chain reaction (RT-PCR). Data obtained from MTT test were analysed using one-way anova, and Tukey's multiple-comparison test. The paired Student t test was employed to compare the effects of materials on gene expression (significance level of 5%). RESULTS: The group cell viabilities were Calcimol LC 53%, Life 43%, TheraCal LC 78%, ProRoot MTA 75% and Biodentine 78%. Calcimol LC and Life exhibited significant differences compared with the control groups (P < 0.05). The percentages of necrotic/late apoptotic cells associated with Calcimol LC and TheraCal LC were greater than in the other materials. However, when RSV was added to wells containing materials, cell viability increased to Calcimol LC 63%, Life 52%, TheraCal LC 82%, ProRoot MTA 91% and Biodentine 96%, and the percentages of early apoptotic and late apoptotic/necrotic cells decreased. Calcimol LC + RSV and Life + RSV differed significantly from the control group (P < 0.05). The expression of IL-8 gene was high for all materials, ProRoot MTA caused significant overexpression, and the addition of RSV reduced the expression of IL-8 in the Calcimol LC, TheraCal LC and ProRoot MTA groups and led to increased expression of IL-10 in the Calcimol LC, Life and Biodentine groups. HBD-2 and BCL-2 exhibited increased expression in ProRoot MTA with RSV (P < 0.05). CONCLUSIONS: The addition of RSV exerted a protective effect on MSCs and regulated the inflammatory process by altering the expression levels of pro- and anti-inflammatory genes.

The role of maternal screening in diagnosing congenital cytomegalovirus infections in highly immune populations.

BACKGROUND: Congenital cytomegalovirus (CMV) infection is the most important infectious cause of central nervous system disease and hearing loss in children. AIMS: In this study, we aimed to investigate the role of maternal screening in early diagnosis of congenital infection in highly immune populations. METHODS: A total of 163 women were included in the study; 103 of the subjects were pregnant and were at full term. The other 60 women were not pregnant, and all were healthy. RESULTS: CMV IgG seropositivity among the pregnant and control groups was found to be 98.1% (101/103) and 98.3% (59/60), respectively, and high IgG avidity was found in all women who had IgG positivity. We did not find any primary CMV infection in the two groups. The recurrent infection rate was found to be 5.82% in the pregnant group and 3.33% in the control group. There were no significant differences between the pregnant and control women in terms of CMV excretion in urine samples (4.85 vs. 3.33%, respectively; P = 1.000) or CMV-DNA presence in serum samples (1.94 vs. 0.0%, respectively; P = 0.532). The presence of symptomatic infection was not observed in any of the 104 babies born from the 103 pregnancies. CONCLUSIONS: According to our results, a maternal screening-based approach would be useful for only a very small group that is at risk of primary infection. Considering the cost of the scan, a routine maternal-based screening program is unadvisable in developing societies, but it is necessary for studies of different cohort groups and infectious diseases.

The role of dental pulp cells in resorption of deciduous teeth.

OBJECTIVE: To address the question whether dental pulp cells of exfoliating human deciduous teeth have some roles for controlling or regulating the root resorption via secreting key molecules (OPG, RANKL, CSF-1, TGFbeta, MCP-1 and Cbfa-1) in osteoclastogenesis, we used a sensitive reverse transcriptase polymerase chain reaction (RT-PCR) method for detection of mRNA expressions for the cytokines listed. STUDY DESIGN: The dental pulps were retrieved from incisor and molar teeth in the late stage of shedding (n = 30) and from sound premolar teeth extracted for orthodontic reasons (control group; n = 30). The RT-PCR assays were used to identify targeted gene expression. RESULTS: Of the cytokines examined, RANKL and CSF-1 expressions showed significantly higher occurrence in deciduous dental pulps than in permanent teeth pulpal tissues (P < .040). CONCLUSIONS: The findings may suggest an interactive role for pulp tissue cells in the physiologic root resorption process. The cells of dental pulp may have some cytokine-producing cells which mediate monocyte-macrophage lineage to form osteo/odontoclasts, and the RANKL/RANK system might be involved in human deciduous teeth resorption.

Detection and quantification of herpesviruses in Kostmann syndrome periodontitis using real-time polymerase chain reaction: a case report.

BACKGROUND/AIMS: Kostmann syndrome, or severe congenital neutropenia, is an autosomal recessive disease of neutrophil production and is associated with severe periodontal pathology. The aim of this study was to determine whether human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV) contribute to the pathogenesis of Kostmann syndrome periodontitis. METHODS: Supragingival plaque and saliva samples were taken from a 6-year-old boy and his 3-year-old sister suffering from Kostmann syndrome, and from two age- and gender-matched healthy children serving as controls. The samples were taken before and 24 months after periodontal treatment. Real-time polymerase chain reaction (TaqMan Real-Time PCR) assay was used to quantify HCMV and EBV DNA. RESULTS: EBV was detected in baseline samples from the Kostmann syndrome patients but not in samples from the healthy control subjects. HCMV was only detected in the saliva of the boy with Kostman syndrome at baseline. Herpesviruses numbers decreased dramatically in the post-treatment samples. CONCLUSION: EBV and HCMV were detected in the two subjects with Kostmann syndrome periodontitis. The results of the study indicate that nonsurgical treatment of Kostmann syndrome periodontitis can reduce supragingival and salivary herpes viral loads.

Human cytomegalovirus, Epstein-Barr virus and bone resorption-inducing cytokines in periapical lesions of deciduous teeth.

BACKGROUND: A connection of herpesvirus periapical infection with symptomatic and large-size periapical lesions has been recognized in adult patients, but no data exist about a possible involvement of herpesviruses in severe periapical pathosis in children. Herpesviruses have the potential to elicit potent bone resorption-inducing cytokines in mammalian cells. AIM: This study aimed to determine the occurrence of human cytomegalovirus and Epstein-Barr virus DNA, and mRNA transcripts of receptor activator of nuclear kappa B ligand (RANKL), osteoprotegerin, core binding factor alpha-1, colony stimulating factor-1, transforming growth factor-beta, and monocyte chemoattractant protein-1 in periapical symptomatic pathosis of deciduous teeth. MATERIAL AND METHODS: Twelve deciduous molar teeth from patients aged 2-8 years were extracted due to severe periapical infection, and granulomatous tissue adherent to the root tip of the extracted teeth was collected using a surgical knife. Non-diseased pulpal tissue, obtained from 12 teeth extracted for orthodontic reasons, served as negative control. Polymerase chain reaction assays were employed to identify herpesvirus DNA and cytokine gene expression, using established polymerase chain reaction primers and procedures. RESULTS: Seven (58%) of the periapical lesions yielded human cytomegalovirus and eight (67%) Epstein-Barr virus. Only one (8%) periapical lesion showed neither human cytomegalovirus nor Epstein-Barr virus. In healthy pulpal tissue, one (8%) specimen demonstrated human cytomegalovirus and another (8%) specimen revealed Epstein-Barr virus. Of the cytokines examined, RANKL expression showed significantly higher occurrence in periapical pathosis than in healthy pulpal tissue (P < 0.040). No relationship was identified between the type of herpesvirus and cytokine expression in the periapical lesions studied. CONCLUSIONS: The present findings provide evidence of a putative role of human cytomegalovirus and Epstein-Barr virus in the pathogenesis of symptomatic periapical pathosis in deciduous teeth. Increased RANKL expression in periapical lesions may be of pathogenetic significance.

Human cytomegalovirus and Epstein-Barr virus type 1 in periodontal abscesses.

OBJECTIVES: Recent studies have linked herpesviruses to severe types of periodontal disease, but no information exists on their relationship to periodontal abscesses. The present study determined the presence of human cytomegalovirus (HCMV) and Epstein-Barr virus type 1 (EBV-1) in periodontal abscesses and the effect of treatment on the subgingival occurrence of these viruses. MATERIAL AND METHODS: Eighteen adults with periodontal abscesses participated in the study. Subgingival samples were collected from each patient with sterile curettes from an abscess-affected site and a healthy control site. HCMV and EBV-1 were identified by polymerase chain reaction at the time of the abscess and at 4 months after surgical and systemic doxycycline therapy. RESULTS: HCMV was detected in 66.7% of periodontal abscess sites and in 5.6% of healthy sites (P=0.002). EBV-1 occurred in 72.2% of abscess sites but not in any healthy site (P<0.001). HCMV and EBV-1 co-infection was identified in 55.6% of the abscess sites. Posttreatment, HCMV and EBV-1 were not found in any study site. CONCLUSIONS: HCMV and EBV-1 genomes are commonly found in periodontal abscesses. These data favor a model in which a herpesvirus infection of the periodontium impairs the host defense and serves as a platform for the entrance of bacterial pathogens into gingival tissue with subsequent risk of abscess development.

Evaluation of the effects of different biomaterials on bone defects.

Studies concerning natural and synthetic graft materials that have been used in different medical procedures have focused on freeze-dried bone, coral, hydroxylapatite, and tricalcium phosphate. This study histologically investigates the effects of these materials on the healing of bone defects. The experiments were performed on 30 albino rabbits. Cavities were drilled in the posterior right tibias of rabbits and were filled with coral, freeze-dried bone, hydroxylapatite, or calcium hydroxide. One cavity was left unfilled as a control. The bone in which the materials were implanted was excised at 7, 15, 30, 45, and 60 days. After the histological staining procedures, the prepared materials were observed using a light microscope. Although all materials showed good bone remodeling at the end of 60 days, coral and hydroxylapatite materials could be seen in the bone structure. The most effective materials within bone defect improvement were freeze-dried bone and calcium hydroxide.