Association between Epstein-Barr virus and periodontitis: A meta-analysis.

PURPOSE: Previous studies have found that Epstein-Barr virus (EBV) is associated with periodontitis, though some controversy remains. This meta-analysis aimed to clarify and update the relationship between EBV and periodontitis as well as clinical parameters. METHODS: A comprehensive search was conducted in the PubMed and Scopus databases in December 2020. Original data were extracted according to defined inclusion and exclusion criteria. Outcomes were analyzed, including overall odds ratios (ORs) and 95% confidence intervals (CIs). A random-effects model was used, and publication bias was assessed by Egger's and Begg's tests. Sensitivity analysis was used to evaluate the stability of the outcome. RESULTS: Twenty-six studies were included in the present meta-analysis, involving 1354 periodontitis patients and 819 healthy controls. The included studies mostly showed high quality. The overall quantitative synthesis for the association between EBV and periodontitis was an increased odds ratio when subgingival EBV was detected OR = 7.069, 95% CI = 4.197-11.905, P<0.001). The results of subgroup analysis suggested that the association of EBV with periodontitis was significant in Asian, European, and American populations (P<0.001; P = 0.04; P = 0.003, respectively) but not in African populations (P = 0.29). Subgroup analysis by sample type showed that subgingival plaque (SgP), tissue and gingival crevicular fluid GCF were useful for EBV detection (P<0.001). EBV detection amplification methods included nested PCR, multiplex PCR and PCR (P<0.001; P = 0.05, P<0.001, respectively), but EBV detection by real-time PCR and loop-mediated isothermal amplification presented no significant result (P = 0.06; P = 0.3, respectively). For the clinical parameters of periodontitis, pocket depth (PD) and bleeding of probing (BOP) percentages were higher in the EBV-positive sites than in the EBV-negative sites (MD 0.47 [0.08, 0.85], P = 0.02; MD 19.45 [4.47, 34.43], P = 0.01). CONCLUSIONS: A high frequency of EBV detection is associated with an increased risk of periodontitis. The EBV association was particularly significant in all populations except in African populations. Subgigival plaque (SgP), tissue and GCF were not significantly different useful material for detecting EBV in periodontitis. Nested PCR and multiplex PCR are reliable methods for this purpose. In the presence of EBV, PD and BOP are reliable clinical parameters for gingival inflammation. However, some caution in such interpretation is justified due to heterogeneity among studies. A suggested extension could assess the parallel influence of other human herpesviruses.

SARS-CoV-2 Detection in Gingival Crevicular Fluid.

Understanding the pathophysiology of the coronavirus disease 2019 (COVID-19) infection remains a significant challenge of our times. The gingival crevicular fluid being representative of systemic status and having a proven track record of detecting viruses and biomarkers forms a logical basis for evaluating the presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The study aimed to assess gingival crevicular fluid (GCF) for evidence of SARS-CoV-2 in 33 patients who were deemed to be COVID-19 positive upon nasopharyngeal sampling. An attempt was also made to comparatively evaluate it with saliva in terms of its sensitivity, as a diagnostic fluid for SARS-CoV-2. GCF and saliva samples were collected from 33 COVID-19-confirmed patients. Total RNA was extracted using NucliSENS easyMAG (bioMérieux) and eluted in the elution buffer. Envelope gene (E gene) of SARS-CoV-2 and human RNase P gene as internal control were detected in GCF samples by using the TRUPCR SARS-CoV-2 RT qPCR kit V-2.0 (I) in an Applied Biosystems 7500 real-time machine. A significant majority of both asymptomatic and mildly symptomatic patients exhibited the presence of the novel coronavirus in their GCF samples. Considering the presence of SARS-CoV-2 RNA in the nasopharyngeal swab sampling as gold standard, the sensitivity of GCF and saliva, respectively, was 63.64% (confidence interval [CI], 45.1% to 79.60%) and 64.52% (CI, 45.37% to 80.77%). GCF was found to be comparable to saliva in terms of its sensitivity to detect SARS-CoV-2. Saliva samples tested positive in 3 of the 12 patients whose GCF tested negative, and likewise GCF tested positive for 2 of the 11 patients whose saliva tested negative on real-time reverse transcription polymerase chain reaction. The results establish GCF as a possible mode of transmission of SARS-CoV-2, which is the first such report in the literature, and also provide the first quantifiable evidence pointing toward a link between the COVID-19 infection and oral health.

[Assessment of measles diagnostic from gingival fluid in Côte d'Ivoire]. / Évaluation du diagnostic de la rougeole à partir des fluides gingivaux en Côte d'Ivoire.

After the adoption of the Global Initiative for Measles Elimination in 2001, Côte d'Ivoire has created monitoring case by case. Thus, the diagnosis of measles from the gingival fluid was implemented, through a pilot project. This study aimed to evaluate the performance of this diagnostic test. We conducted a cross-sectional survey, in four health districts of Abidjan, during a period from July 2010 to December 2012. The study consisted in collecting gingival fluid and serum samples in all suspected measles children. These samples were analyzed by ELISA test at Pasteur Institute of Côte d'Ivoire. Standard formulas were used to calculate sensitivity, specificity, positive predictive value and negative predictive value of oral fluid compared to serum taken as the "gold standard" and confidence intervals were estimated with error alpha risk (α =0.05). The concordance of kappa coefficient (k) was used to estimate agreement level between the results of oral fluid analysis and those of serum. The sensitivity and specificity of the test were 98% and 82% respectively while the positive predictive value and the negative predictive value were 84% and 98%. The comparison of oral fluid with the reference test showed high agreement, between 0.61 and 0.80. The diagnostic test on gingival fluid is acceptable because its sensitivity, specificity and negative predictive value had high level. Therefore it can be extended to all sanitary districts.

THE PERFORMANCE OF A RAPID TEST FOR ANTI-HCV SCREENING IN ORAL FLUIDS.

UNLABELLED: Gingival crevicular fluid (GCF) and saliva samples provide advantages for screening or sero-prevalence studies on HCV using less invasive methods. The study aimed to evaluate the performance of a rapid test for HCV-antibodies (HCV-Ab) screening in oral fluids among high-risk individuals with chronic liver disease. METHODS: Chronic liver disease patients attending at the Matei Bals National Instiute for Infectious Diseases were recruited for this study. Plasma, GCF and saliva samples (pair samples) were collected from each patient included in the study. Forty-three sample pairs were tested with Laboquick (Koroglu Medical Devices) rapid test and ELISA (DIA.PRO--Diagnostic Bio-probes) for the detection of anti-HCV antibodies. RESULTS: Using rapid test, anti-HCV antibodies were detected in 36 GCFs (83.72%) and 24 saliva cases (55.8%) of infected subjects. For a better estimation of oral fluids positivity, the cut-off values were calculated following plotting the ROC curves (COV2). Comparing Laboquick and ELISA (COV2) data, matched results were noted in 95.3 % saliva samples and 93% GCF samples. CONCLUSIONS: Oral fluids could be an alternative to blood for detection of HCV-positive subjects. Anti-HCV rapid test may be useful in routine dental medicine.

Detection of herpes simplex virus type 1 in gingival crevicular fluid of gingival sulcus/periodontal pocket using polymerase chain reaction.

INTRODUCTION: Pathogenesis and some characteristics of periodontitis cannot be fully explained by bacterial etiology alone. Herpes viruses may bridge the gap between clinical characteristics and molecular understanding of periodontal destruction. OBJECTIVE: The aim of this study was to investigate the prevalence of herpes simplex virus type 1 (HSV-1) in gingival crevicular fluid (GCF) of healthy and damaged periodontium in Serbian population and to explore potential correlation between the presence of this virus and the level of periodontal destruction. METHODS: Samples were collected from gingival sulcus/periodontal pockets by sterile paper points and the presence of viral DNA in gingival crevicular fluid was assessed by PCR. RESULTS: There was no statistically significant difference in HSV-1 in presence between periodontitis patients (PG = 38.9%) and healthy controls (HC = 32.3%), (Chi-square test, with Yates' correction p = 0.7574). However, HSV-1 positive patients showed significantly higher values of parameters of periodontal destruction (PPD = 7.11 +/- 2.52, CAL = 5.46 +/- 2.34) than periodontitis patients without HSV-1 in gingival crevicular fluid (PPD = 4.70 +/- 1.79, CAL = 3.39 +/- 2.65) (p values respectively, p = 0.002 and p = 0.023, Independent Samples T-Test). HSV-1 occurred more often in deeper (PPD > or = 6 mm) (69.2%) than in shallow pockets (3 mm < PPD < 6 mm) (18.2%) (Chi-square test, with Yates' correction, p = 0.008). Plaque index was lower in the HSV-1 positive group (0.84 +/- 0.69 vs. 1.43 +/- 0.76, p = 0.023, Independent Samples T-Test). CONCLUSION: This study demonstrated that the presence of HSV-1 in the gingival crevicular fluid coincides with a higher degree of tissue destruction in patients with periodontitis.

Oral fluid and hepatitis A, B and C: a literature review.

BACKGROUND AND AIMS: Viral hepatitis is a significant global health problem that, depending upon the virus, affects individuals of the developing and/or developed world. In recent years, there has been renewed interest in whether oral fluids can be considered as a source of viral hepatitis transmission and whether oral fluid, in particular, whole saliva, may be a useful source for viral detection as part of the diagnosis and monitoring of viral hepatitis. The aim of this article was to review current data concerning the possible carriage of the hepatitis A, B and C viruses within saliva and gingival crevicular fluid. Such knowledge will indicate if (i) oral fluid is a possible source of infection and (ii) whether oral fluid can be used for diagnosis and monitoring of viral hepatitis. DATA AND SOURCES: A literature search was conducted using PubMed (Medline), EMBASE/Excerpta medica, the Cochrane database and Scopus. The results were limited to published material after 2000. Relevant material was evaluated and reviewed. CONCLUSION: There is some evidence that hepatitis viruses A, B and C are present in oral fluids, particularly whole saliva and gingival crevicular fluid and may thus be possible sources of viral detection in clinical diagnosis and monitoring. However, the data are inconsistent and warrant the need for well-planned longitudinal studies to explore the precise frequency of oral carriage of such viruses and to determine the virological and host factors that may influence the oral presence of hepatitis A, B and C viruses.

[The relationship between two kinds of human herpesviruses and experimental gingivitis].

OBJECTIVE: To investigate human cytomegalovirus (HCMV) and Epstein-Barr virus-type 1 (EBV-1) in GCF and saliva during experimental gingivitis in Chinese young subjects and to evaluate the effect of the virus in the initial stage of gingival inflammation. METHODS: GCF of 14 and 45 and saliva without stimulating in 11 Chinese young males with healthy gingiva were collected at baseline (day 0), day 7, 14 and 21 after stopping oral hygiene and day7 after reestablishing oral hygiene (day 28). DNA of HCMV and EBV-1 were detected by nested-polymerase chain reaction (n-PCR) at the times mentioned above. RESULTS: HCMV was detected in GCF of 4 subjects at baseline, 4 subjects at day 7, 3 subjects at day 14 and 2 subjects at day 21 while the subjects were different. At day 28 HCMV could not be detected. EBV-1 was not detectable in GCF during the experimental gingivitis. HCMV was detected in saliva in 4 subjects and EBV-1 was in 3 subjects. And there is no relationship between the detection of the herpesviruses and the clinical parameters as well. CONCLUSION: We suggest that HCMV and EBV-1 are not the important factors during the initial stage of gingival inflammation.

[Detection of human papillomavirus in gingival fluid of Venezuelan HIV patients with periodontal disease]. / Detección de virus papiloma humano en el fluido gingival de pacientes con inmunodeficiencia humana y enfermedad periodontal.

Evidence suggests that viruses may be involved in the activation of periodontal disease, allowing the overgrowth of periodontal pathogens. The purpose of the present study was to detect the presence of Human Papillomavirus (HPV) in gingival crevicular fluid (GCF) in HIV+ Venezuelan patients with periodontal disease. We evaluated GCF samples from 20 HIV+ patients with periodontal disease from the Infectious Disease Center, Faculty of Dentistry, Central University of Venezuela, and were clinically examined to establish their periodontal conditions, 13 under HAART (antiretroviral therapy) and 7 without HAART. Seven seronegative patients with chronic periodontitis and 7 seronegative patients, without periodontal disease were included. DNA extraction was performed, the consensus primers MY09 and MY11 for the HPV L1 region were used for PCR amplification. Genotipification was made for the 6, 11, 16, 18, 31 and 45 genotypes. HPV were detected in 46% of HIV+ patients under therapy. The CD4 cell counts in the IIPV+ patients were not significantly different from the HPV-group. The viral load in the HPV+ group was significantly higher (200,470 +/- 324,244 copy/mL) than in the HPV-patients (10,246 +/- 23,805 copy/mL). Genotypes 6 and 11 were observed in the HPV positive samples, of which 4/6 (66.6%) presented coinfection with both types. No significant differences in the periodontal conditions were observed between patients with IIPV-HIV infection related to patients with only HIV. HPV was detected only in the gingival crevicular fluid of HIV+ patients under HAART independently of the periodontal conditions.

Detección de Virus Papiloma Humano en el fluido gingival de pacientes con inmunodeficiencia humana y enfermedad periodontal / Detection of Human Papillomavirus in gingival fluid of Venezuelan HIV patients with periodontal disease

Evidencias sugieren que los virus pueden participar en la activación de la enfermedad periodontal, permitiendo el sobrecrecimiento de bacterias periodontopatógenas. El objetivo del estudio fué la detección molecular de VPH en fluido gingival (FG) de pacientes VIH+ con enfermedad periodontal. Se evaluaron muestras de FG de 20 pacientes VIH+ con enfermedad periodontal que asistieron al Centro de Atención de Pacientes con Enfermedades Infecciosas (CAPEI) de la Facultad de Odontología de la Universidad Central de Venezuela, 13 bajo terapia antirretroviral (HAART) y 7 VIH+ sin HAART. Se incluyeron 7 pacientes seronegativos con periodontitis crónica y como grupo control 7 pacientes seronegativos periodontalmente sanos. Se extrajo el ADN, se amplificó la región L1 de VPH con primers MY09 y MY11. Las muestras VPH+ fueron genotipificadas para los tipos 6, 11, 16, 18, 31 y 45. VPH fue detectado en 46% de los pacientes VIH+ bajo terapia. El contaje CD4+ en la población VPH+ no presentó diferencias con el grupo VPH-, y la carga viral mostró valores promedio significativamente mayores (200.470± 324.244 copias/mL) con respecto a los pacientes VPH- (10.246±23.805 copias/mL). Las muestras VPH+ presentaron los genotipos 6 y 11, de los cuales 66,6% estaban coinfectados con ambos tipos. Las condiciones periodontales no presentaron diferencias entre los individuos con doble infección viral por VPH y VIH, y los que solo portaban VIH. VPH fue detectado solamente en fluido gingival de pacientes VIH+ con HAART, indicando que esta terapia puede influir en el estado inmunológico independientemente de las condiciones periodontales.

Evidence suggests that viruses may be involved in the activation of periodontal disease, allowing the overgrowth of periodontal pathogens. The purpose of the present study was to detect the presence of Human Papillomavirus (HPV) in gingival crevicular fluid (GCF) in HIV+ Venezuelan patients with periodontal disease. We evaluated GCF samples from 20 HIV+ patients with periodontal disease from the Infectious Disease Center, Faculty of Dentistry, Central University of Venezuela, and were clinically examined to establish their periodontal conditions, 13 under HAART (antiretroviral therapy) and 7 without HAART. Seven seronegative patients with chronic periodontitis and 7 seronegative patients, without periodontal disease were included. DNA extraction was performed, the consensus primers MY09 and MY11 for the HPV L1 region were used for PCR amplification. Genotipification was made for the 6, 11, 16, 18, 31 and 45 genotypes. HPV were detected in 46% of HIV+ patients under therapy. The CD4 cell counts in the HPV+ patients were not significantly different from the HPV-group. The viral load in the HPV+ group was significantly higher (200,470 ± 324,244 copy/mL) than in the HPV- patients (10,246 ± 23,805 copy/mL). Genotypes 6 and 11 were observed in the HPV positive samples, of which 4/6 (66.6%) presented coinfection with both types. No significant differences in the periodontal conditions were observed between patients with HPV-HIV infection related to patients with only HIV. HPV was detected only in the gingival crevicular fluid of HIV+ patients under HAART independently of the periodontal conditions.

[Effect of periodontal mechanical treatment on herpesviruses in gingival crevicular fluid of patients with chronic periodontitis].

OBJECTIVE: To evaluate the subgingival prevalence of human cytomegalovirus (HCMV), Epstein-Barr virus-1 (EBV-1) in chronic periodontitis (CP) patients before and after treatment and to analyze the relationship between the prevalent variance and periodontal clinical parameters. METHODS: Gingival crevicular fluids of 13 CP patients were collected at baseline, 2 weeks, 2 months and 4 months after periodontal mechanical treatment. HCMV and EBV-1 were detected using nested polymerase chain reaction (n-PCR). RESULTS: The plaque index (PLI), probing depth (PD) and bleeding index (BI) of CP patients at 2 months, 4 months after periodontal mechanical treatment were evidently lower than before treatment, P < 0.01. These parameters at 4 months after treatment were higher than at 2 months, the differences were significant, P < 0.05. The prevalence of HCMV and EBV in CP patients was 42% (33/78), 14% (11/78). EBV and HCMV were mostly coexistent in the same site [9 sites HCMV(+) in 11 EBV positive sites]. The sites of HCMV(+) and EBV(+) were almost deep pockets. Thirteen of 14 sites with deep pockets were HCMV(+), 9 sites were deep pockets in 11 sites EBV(+). The prevalence of HCMV and EBV (8% and 0 respectively) at 2 weeks was the lowest in all four time points. The prevalence of HCMV and EBV at 2 weeks, 2 months and 4 months following treatment was significantly lower than baseline (P < 0.01), but the prevalence of HCMV (15%) at 2 months after treatment was higher than at 2 weeks (8%), the difference was not significant (P = 0.133). CONCLUSIONS: Herpesviruses may play a role in the development of CP. The changes of the prevalence of herpesviruses before the changes of clinical parameters could be detected after periodontal mechanical treatment. The patients should be re-evaluated and re-treated within 2 months after treatment.

Detection of herpetic viruses in gingival crevicular fluid of patients suffering from periodontal diseases: prevalence and effect of treatment.

BACKGROUND/AIM: Although the role of bacteria in the etiology of periodontitis is well established, it has been suggested that herpetic viruses could contribute to the initiation and progression of this disease. The aim of this study was to determine the prevalence of human cytomegalovirus (HCMV), Epstein-Barr virus (EBV) and herpes simplex virus (HSV) in gingival crevicular fluid (GCF) samples obtained from periodontally healthy, gingivitis and periodontitis patients. In addition, the effect of periodontal treatment (scaling and root planing) on the persistence of herpetic viruses was evaluated in a sub-group of patients suffering from chronic periodontitis. METHODS: The presence of viruses in GCF samples was assessed by a nested PCR amplification technique. The persistence of viruses in periodontal sites was evaluated following a scaling and root planing therapy. RESULTS: A statistically significant higher prevalence of HCMV was observed in periodontitis patients as compared to healthy control subjects (35 vs. 8%, respectively; P = 0.0377). A trend for a higher prevalence of HSV was also noted in the periodontitis group, in comparison with healthy control subjects. In addition, a higher prevalence of HCMV was associated with deep periodontal pockets in subjects suffering from periodontitis. In the sub-group of periodontitis patients, periodontal therapy resulted in the elimination (HCMV and EBV) or reduction (HSV) of the herpetic viruses. CONCLUSIONS: This study showed that the prevalence of HCMV and HSV viruses in GCF is higher in patients suffering from periodontitis compared to periodontally healthy subjects, and that the prevalence of HCMV is higher in deep periodontal pockets. It also brought evidences that periodontal therapy may be associated with virus elimination in diseased sites.

Salivary levels of Epstein-Barr virus DNA correlate with subgingival levels, not severity of periodontitis.

OBJECTIVE: The aim of this study was to determine the presence and quantity of human cytomegalovirus (CMV) and Epstein-Barr virus (EBV) DNA in the saliva of patients with periodontitis, and investigate the correlation between these factors. METHODS: Presence and amounts of viral DNA in saliva and subgingival plaque samples, from healthy and disease sites, of 65 adults diagnosed with chronic periodontitis were determined using quantitative real-time polymerase chain reaction. RESULTS: Epstein-Barr virus DNA was detected in saliva of 81.5% (53/65) of patients at a median concentration of 4325 copies ml(-1). CMV DNA was detected in saliva of one individual (1.5%) at low copy number. Patients who had EBV in saliva were 10 times more likely to have EBV in subgingival plaque than patients lacking EBV in saliva (odds ratio = 10.1, 95% confidence interval = 2.6-39.5; P = 0.0009). EBV DNA burden in saliva positively correlated with the amounts detected in plaque and with amounts detected in increasing number of affected sites (P < 0.0001). EBV DNA presence and quantity in saliva did not correlate with increasing severity of disease as measured by periodontal indices. CONCLUSIONS: Epstein-Barr virus DNA presence and burden in saliva are associated with its presence and burden in subgingival plaque, but presence and burden in saliva does not correlate with periodontal disease severity.

Comparison of nested polymerase chain reaction (PCR), real-time PCR and viral culture for the detection of cytomegalovirus in subgingival samples.

INTRODUCTION: The purpose of this study was to compare nested polymerase chain reaction (PCR), real-time PCR, and shell vial for the detection of human cytomegalovirus (HCMV) in subgingival samples in periodontitis patients. METHODS: A group of 44 patients and 24 individuals without periodontitis were included in the study. A full periodontal examination was conducted in each subject. Gingival crevicular fluid (GCF) was collected by pocket lavage and used for viral culture (shell vial). Additional subgingival samples were obtained with paper points and used for molecular analysis. Nested PCR and real-time PCR were used to detect and quantify HCMV. Student's t-test and chi-squared test were used to compare groups. The sensitivity and specificity for the tests were calculated on 2 x 2 tables considering the nested PCR as the gold standard. RESULTS: The detection of HCMV was greater using nested PCR than with either real-time PCR or shell vial (P < 0.0001). However, the frequency detection of both molecular techniques was higher than in viral culture (P < 0.0001). Only one case of chronic periodontitis was positive by viral culture. Agreement between nested PCR and real-time PCR was observed 47.7% and 4.1% of the time in the periodontitis and control groups, respectively. The sensitivity of real-time PCR was 60%, compared with 2.8% for the shell vial technique. CONCLUSIONS: In conclusion, this study confirmed that active HCMV infection occurs in human periodontitis; however, its frequency seems to be low. In contrast, latent periodontal HCMV infection seems to be a more frequent event.

Microbiology and cytokine levels around healthy dental implants and teeth.

BACKGROUND: Elicitation of the relationship of periodontopathogens and pro-inflammatory cytokines to bone resorption and formation is significant to a growing body of research known as osteoimmunology. It is essential that clinically healthy peri-implant and periodontal sites are studied to contribute comparison data for investigations that are addressing diseased sites. PURPOSE: The purpose of this study was to describe levels of selected pro-inflammatory cytokines in clinically healthy peri-implant and periodontal sites, and to examine whether cytokine levels may be related to specific bacterial/viral pathogens. MATERIALS AND METHODS: Eleven subjects (mean age 56.2 +/- 10) participated in the study. Subgingival microbial samples were cultured for periodontopathic bacteria. Gingival crevicular fluid samples were analyzed by nested polymerase chain reaction for Cytomegalovirus (HCMV) and were tested for the quantification of Interleukin (IL)-8, IL-1beta, IL-6, IL-10, Tumor Necrosis Factor (TNF)-alpha, and IL-12p70 using flow cytometry (FACS). Findings for microbiota composition and cytokine levels were compared between implants and teeth (chi square, Kruskall-Wallis, Mann-Whitney; p < or = .05). RESULTS: Both the frequency (%) and levels (%) of periodontopathic bacteria were higher around teeth than implants. The concentration (picogram per milliliter) of cytokines was more prominent around implants than teeth, reaching nearly twofold differences in some instances. Cytokine levels were higher when the sites analyzed were positive for any bacteria tested. HCMV was not detected. CONCLUSIONS: Pro-inflammatory cytokine production was unrelated to heavy bacterial challenge. Nevertheless, when periodontopathic bacteria were detected by culture, cytokine levels were increased around both implants and teeth. Studies are needed to investigate the pro-inflammatory cytokines (especially IL-1beta and TNF-alpha) produced in spite of minimal bacterial accumulation.

Immunology, microbiology, and virology following placement of NobelPerfect scalloped dental implants: analysis of a case series.

BACKGROUND: Cytokine-microbiology-virology monitoring after implant placement may help to develop profiles of variables that can help to explain interaction between the immune system and alveolar bone. Descriptive information at the molecular and cellular levels after implant placement is important in the emerging field of osteoimmunology and may help to formulate hypotheses and intervention strategies in periodontology and implantology. PURPOSE: The purpose of this study was to determine the presence or absence of selected cytokines in association with periodontopathogens and human cytomegalovirus (HCMV) after placement of dental implants. MATERIALS AND METHODS: Charts of seven consecutive patients with 19 NobelPerfect (Nobel Biocare, Yorba Linda, CA, USA) implants were reviewed for crevicular fluid sample outcomes. Anaerobic culture determined periodontopathogens 2 to 5 days, 3 and 6 months postimplant insertion. At 3, 6, and 12 months, real-time reverse transcriptase-polymerase chain reaction (RT-PCR) was performed to detect active HCMV, interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), and interferon-gamma (INF-gamma). RESULTS: Four of five, and six of seven patients harbored no periodontopathogens at 3- or 6-month intervals, respectively. In spite of absence of a bacterial challenge, IL-1beta and TNF-alpha activity was significant. INF-gamma was not detected, and HCMV was present at one time interval only. CONCLUSIONS: TNF-alpha is produced mainly in the early stages of acute inflammation, and high levels of this cytokine at 3 and 6 months postimplant placement may be related to a repetitive acute-phase inflammatory response. Lack of INF-gamma and a high cytokine presence without significant corresponding periopathogens or viruses raise a concern that inflammation and, thus, inflammatory bone destruction, is possible outside of these variables. Inflammation and bone loss around this same group of scalloped implants, reported by our previous study, may have been initiated by local factors, such as particular implant design features.

The association between human herpes viruses and periodontal disease, part 2.

The role of herpesviruses in the aetiopathogenesis of periodontal diseases is uncertain. While their role in the initiation and progression of periodontal disease has not been established, the increased levels of herpesvirus gene products in periodontally diseased sites--compared to that in clinically healthy sites--and their association with elevated levels of periodontopathic bacteria points to a possible coactive role of the herpesviruses with the bacteria. The available data regarding the relationship between herpesviruses and the periodontal disease status, as well as the possible mechanisms that might enable herpesviruses to influence the pathogenesis of periodontal diseases, are discussed.

Detection of human herpesvirus 6 in patients with oral chronic graft-vs-host disease following allogeneic progenitor cell transplantation.

INTRODUCTION: Chronic graft-vs-host disease (cGVHD) is a major cause of morbidity in long-term survivors of allogeneic hematopoietic progenitor cell transplantation. Herpesviruses are involved in the occurrence and progression of various oral diseases. AIM: The aim of this study was to investigate the role of human herpesvirus 6 (HHV6) in patients with oral manifestations of cGVHD. MATERIALS AND METHODS: Peripheral blood and oral fluids (whole saliva, gingival crevicular fluid and parotid gland saliva) from 19 cGVHD patients, and 28 blood donors were examined for HHV6. Oral tissue samples were collected from 12 cGVHD patients and 12 healthy individuals. Nested polymerase chain reaction was employed to identify the HHV6. RESULTS AND CONCLUSION: The virus was detected in whole saliva in 13 cGVHD patients (68%) and in 19 blood donors (67%). HHV6 was not identified in any of the gingival crevicular fluid and parotid gland saliva samples in cGVHD patients. In the control group 14.3% of both, four gingival crevicular fluid and four parotid gland saliva samples were positive. Two oral tissue samples of cGVHD patients were positive for HHV6. These results indicate that patients with oral manifestations of cGVHD and healthy individuals present high and similar incidence of HHV6 in blood and oral fluids. These data do not support the importance of HHV6 in oral lesions of cGVHD.

(B2) Periodontal diseases and other bacterial infections.

The workshop addressed the following questions with respect to periodontal diseases and bacterial infections seen in HIV infection: (1) What is linear gingival erythema? Is it prevalent only in HIV disease? A crude Delphi technique was used to ascertain whether LGE existed, but a consensus could not be reached. It was agreed that a diagnosis of LGE should be considered only if the lesion persists after removal of plaque in the initial visit. (2) Do periodontal pockets contribute to viremia in HIV infection? At present, the data are not available to answer this question. (3) Do anti-viral drugs reach the sulcular fluid in significant concentrations? No one at the workshop was aware of data that could answer this question. (4) Does concurrent tuberculosis infection modify the oral manifestations of HIV infection? Though analysis of data from the developing countries does suggest an association between tuberculosis and oral candidiasis, more data and multivariate analysis considering immunosuppression as a confounding factor are necessary, for any conclusions to be derived. (5) What pathogens are involved in periodontal diseases in HIV infection? Periodontal disease may be initiated by conventional periodontal pathogens. But the progression and tissue destruction depend upon the presence of typical and atypical micro-organisms, including viruses, their by-products, increased secretion of potentially destructive inflammatory mediators, and overwhelming host response. (6) How can we diagnose the diseases seen in HIV infection? The answer can be obtained only with data from controlled and blinded studies. It is necessary to design collaborative multi-center longitudinal studies. The results obtained from such large sample sizes can contribute eventually to interpretation of the outcome.

Quantitative detection of hepatitis C virus (HCV) RNA in saliva and gingival crevicular fluid of HCV-infected patients.

The search for hepatitis C virus (HCV) in body fluids other than blood is important when assessing possible nonparenteral routes of viral transmission. However, the role of oral fluids in HCV transmission remains controversial. Here we quantitatively determined HCV RNA in saliva and gingival crevicular fluid (GCF) of anti-HCV-positive patients. Most patients (14 of 18; 78%) whose saliva specimens were negative had HCV RNA in their GCF. Most patients (20 of 26; 77%) had higher HCV RNA levels in their GCF than in their saliva. Although there was not a statistically significant correlation between the serum viral load and HCV level in saliva or GCF, patients with low serum HCV loads were less likely to have detectable HCV in their saliva. These findings have important implications for medical personnel and suggest that epidemiological studies designed to understand the significance of the oral route of transmission of HCV are warranted.