Role of Candida species from HIV infected children in enamel caries lesions: an in vitro study.

OBJECTIVES: This study analyzed the capacity of Candida spp. from dental biofilm of HIV infected (HIV+) children to demineralize primary molar enamel in vitro by Transversal Microhardness (TMH), Polarized Light Microscopy (PLM) and the quantity of calcium ions (Ca2+) released from the enamel. MATERIAL AND METHODS: Candida spp. samples were isolated from the supragingival biofilm of HIV+ children. A hundred and forty (140) enamel blocks were randomly assigned to six groups: biofilm formed by C. albicans (Group 1); mixed biofilm formed by C. albicans and C. tropicalis (Group 2); mixed biofilm formed by C. albicans and C. parapsilosis (Group 3); mixed biofilm formed by C. albicans, C. parapsilosis and C. glabrata (Group 4); biofilm formed by C. albicans ATCC (Group 5) and medium without Candida (Group 6). Enamel blocks from each group were removed on days 3, 5, 8 and 15 after biofilm formation to evaluate the TMH and images of enamel were analyzed by PLM. The quantity of Ca2+ released, from Groups 1 and 6, was determined using an Atomic Absorption Spectrophotometer. The SPSS program was used for statistical analysis and the significance level was 5%. RESULTS: TMH showed a gradual reduction in enamel hardness (p<0.05) from the 1st to 15th day, but mainly five days after biofilm formation in all groups. The PLM showed superficial lesions indicating an increase in porosity. C. albicans caused the release of Ca2+ into suspension during biofilm formation. CONCLUSION: Candida species from dental biofilm of HIV+ children can cause demineralization of primary enamel in vitro.

Role of Candida species from HIV infected children in enamel caries lesions: an in vitro study

Abstract Objectives This study analyzed the capacity of Candida spp. from dental biofilm of HIV infected (HIV+) children to demineralize primary molar enamel in vitro by Transversal Microhardness (TMH), Polarized Light Microscopy (PLM) and the quantity of calcium ions (Ca2+) released from the enamel. Material and Methods Candida spp. samples were isolated from the supragingival biofilm of HIV+ children. A hundred and forty (140) enamel blocks were randomly assigned to six groups: biofilm formed by C. albicans (Group 1); mixed biofilm formed by C. albicans and C. tropicalis (Group 2); mixed biofilm formed by C. albicans and C. parapsilosis (Group 3); mixed biofilm formed by C. albicans, C. parapsilosis and C. glabrata (Group 4); biofilm formed by C. albicans ATCC (Group 5) and medium without Candida (Group 6). Enamel blocks from each group were removed on days 3, 5, 8 and 15 after biofilm formation to evaluate the TMH and images of enamel were analyzed by PLM. The quantity of Ca2+ released, from Groups 1 and 6, was determined using an Atomic Absorption Spectrophotometer. The SPSS program was used for statistical analysis and the significance level was 5%. Results TMH showed a gradual reduction in enamel hardness (p<0.05) from the 1st to 15th day, but mainly five days after biofilm formation in all groups. The PLM showed superficial lesions indicating an increase in porosity. C. albicans caused the release of Ca2+ into suspension during biofilm formation. Conclusion Candida species from dental biofilm of HIV+ children can cause demineralization of primary enamel in vitro.

Levels of HIV-1 in subgingival biofilm of HIV-infected patients.

AIM: The aims of the current study were to compare the levels of HIV-1 in the subgingival biofilm (SHVL) between detectable and undetectable plasmatic HIV-1 viral load (PHVL) in HIV-infected patients as well as to determine the association of SHVL with PHVL and clinical periodontal parameters. MATERIAL AND METHODS: Forty-one HIV-infected individuals were divided into two groups: detectable (21) and undetectable (20) PHVL. Subgingival biofilm samples were obtained for detection and quantification of HIV-1 by real-time RT-PCR. To estimate the effect of co-variables on the outcome undetectable SHVL, the Generalized Estimation Equation (GEE) was employed. RESULTS: Detectable SHVL was observed only in the detectable PHVL group and the detection of the HIV-1 was observed in 40% of these individuals. In the bivariate analysis between co-variables from the individual level and the outcome SHVL, significant difference was observed only for the CD4+ T lymphocytes levels (p = 0.017). The multiple logistic model demonstrated that only CD4+ T lymphocytes levels had a significant effect on the outcome undetectable SHVL [OR 8.85 (CI 3.6-9.2), p = 0.002]. CONCLUSION: HIV-1 can be detected and quantified in the subgingival biofilm of HIV-infected individuals, but these findings are not associated with PHVL and periodontal clinical parameters.

Associação entre a carga viral plasmática e a carga viral subgengival no biofilme em pacientes infectados pelo HIV-1 / Association between plasmatic viral load and subgingival viral load in biofilm in HIV-1 infected patients

Objetivo: Investigar a associação entre carga viral no biofilme subgengival (CVS) e carga viral plasmática (CVP) do HIV-1 e os parâmetros clínicos periodontais em indivíduos infectados pelo HIV. Metodologia: Quarenta e um pacientes com sorologia positiva para HIV-1 foram distribuídos em dois grupos: 20 com CVP detectável e 21 com CVP indetectável (< 50 cópias/mL). Amostras do biofilme subgengival foram coletadas de 12 sítios de cada paciente com periodontite, sendo 6 sítios com as maiores PBS, preferencialmente não adjacentes, e 6 sítios com periodonto sadio. Nos pacientes com saúde periodontal, foram coletadas 6 amostras de sítios periodontais sadios aleatórios. Das 6 amostras dos sítios com maiores PBS dos pacientes com periodontite, foram realizados 3 pools, com 2 amostras cada, de sítios com parâmetros clínicos semelhantes. O mesmo foi feito para as 6 amostras de periodontos sadios (tanto para os pacientes com periodontite como para aqueles com saúde periodontal), resultando em 3 pools. A detecção e quantificação da CVS foram realizadas através do PCR em Tempo Real. Os níveis de linfócitos T CD4+, T CD8+, neutrófilos e CVP foram obtidos dos resultados desses exames presentes nos prontuários médicos de cada paciente. Diferenças significativas entre os grupos para todos os parâmetros estudados foram avaliadas pelos testes Qui-quadrado, exato de Fisher e Mann-Whitney. O modelo de regressão utilizado foi a Equação de Estimação Generalizada (GEE) para testar a associação entre as co-variáveis e a CVS. Resultados: A maioria dos indivíduos era composta por homens (51,2%), não usuários de drogas (85,4%), não tabagistas (63,4%) e não consumidores diários de álcool (87,8%). Os dados laboratoriais demonstraram diferenças significantes entre os dois grupos estudados somente para os níveis de linfócitos T CD4+ (p = 0,038), porém não houve diferença estatisticamente significante entre os grupos em relação aos parâmetros clínicos periodontais. Não houve associação estatisticamente significante entre CV plasmática e CV subgengival, sendo a contagem de linfócitos TCD4+ a única co-variável que apresentou efeito estatisticamente significante sobre o desfecho CV subgengival indetectável (p = 0,017). A interpretação dos resultados demonstrou que indivíduos com níveis mais altos de linfócitos T CD4+ (> 500 células/mm3 ) tem uma chance muito maior de apresentar CV subgengival indetectável em relação aos indivíduos com níveis de linfócito T CD4+ < 200 células/mm3 . Conclusão: Não há associação entre a carga viral no biofilme subgengival e a carga viral plasmática do HIV-1 e os parâmetros clínicos periodontais em pacientes infectados pelo HIV.(AU)

Aim: To investigate the association between subgingival viral load in biofilm and plasmatic viral load and clinical periodontal parameters in HIV-1 infected patients. Methods: Fortyone HIV-positive patients were divided into two groups: detectable and undetectable plasmatic viral load (< 50 copies/mL). Subgingival biofilm samples were collected from 12 sites in each patient with periodontitis, 6 sites with the deepest PBS, preferably not adjacent, and 6 sites with healthy periodontium. In patients with periodontal health, 6 samples were collected from random healthy periodontal sites. Of the six samples of periodontal lesions were performed 3 pools, with 2 samples from each site with similar clinical parameters. The same was done with the 6 samples from healthy periodontium, resulting in three pools. Detection and quantification of subgingival viral load were determined by Real Time PCR. The levels of CD4, CD8, neutrophils and plasmatic viral load were obtained from the medical records of each patient. Significant differences between groups for all parameters were evaluated by Chi-square, Fisher's exact test and Mann-Whitney. The utilized regression model consisted of a test called Generalized Estimation Equation (GEE) to test the association between the co-variables and the subgingival viral load. Results: The majority of the patients were male (51,2%), non-drug users (85,4%), non-smokers (63,4%) and nonalcohol users (87,8%). The laboratory data demonstrated significant differences between groups only for levels of TCD4+ cells (p=0,038), however there was no significant difference between groups for clinical periodontal parameters. There was no statistically significant association between plasmatic viral load and subgingival viral load, and only the levels of TCD4+ cells demonstrated significant effect on the undetectable subgingival viral load (p=0,017). The interpretation of the results demonstrated that patients with higher levels of TCD4+ cells (> 500 cells/mm3 ) have a much higher chance of presenting undetectable subgingival viral load compared to patients with levels of TCD4+ cells < 200 cells/mm3. Conclusion: There is no association between subgingival viral load in biofilm and plasmatic viral load and clinical periodontal parameters in HIV-1 infected patients(AU)

Relationship between herpesviruses and periodontopathogens in patients with HIV and periodontitis.

BACKGROUND: The purpose of the present study is to verify a possible association between herpesviruses and periodontal pathogens in individuals with human immunodeficiency virus (HIV) and periodontitis. METHODS: Twenty-seven patients with HIV and chronic periodontitis and 23 patients with HIV and gingivitis were included in the study. Probing depth, clinical attachment loss, gingival index, and plaque index were recorded. Blood, saliva, and subgingival plaque were processed for viral and bacterial identification. Bacteria were identified by 16S rRNA-based polymerase chain reaction and viruses by the nested polymerase chain reaction. RESULTS: For the chronic periodontitis group, Epstein-Barr (EBV)-1 (70.4%) and Tannerella forsythia (Tf) (51.8%) presented higher detection in subgingival plaque and saliva (81.5% and 40.7%, respectively) than in blood (22% and 0%, respectively) (P <0.005 and P <0.0001, respectively). Porphyromonas gingivalis (Pg) was more frequent in subgingival plaque (77.7%; P <0.0001). In the gingivitis group, Pg and human cytomegalovirus (HCMV) presented higher frequency in subgingival plaque (95.6% and 91.3%, respectively; P <0.0001 and P = 0.004). Tf and EBV-1 were detected more frequently in subgingival plaque (47.8% and 78.3%, respectively) and saliva (52.2% and 52.2%, respectively; P = 0.004 and P <0.005) than in blood. EBV-1, EBV-1-HCMV, and presence of different viruses presented an association with periodontitis in saliva. CONCLUSIONS: No association was detected for herpesviruses and periodontal pathogens in patients who are HIV-positive with periodontitis. EBV-1 and coinfection (EBV-1-HCMV) were associated with patients who are HIV-positive with periodontitis.

Detection of herpesviruses and periodontal pathogens in subgingival plaque of patients with chronic periodontitis, generalized aggressive periodontitis, or gingivitis.

BACKGROUND: Herpesviruses may be related to the etiology of aggressive periodontitis (AgP) and chronic periodontitis (CP) by triggering periodontal destruction or by increasing the risk for bacterial infection. This case-control study evaluated the presence of herpes simplex virus type I (HSV-1), Epstein-Barr virus type I (EBV-1), human cytomegalovirus (HCMV), Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans), Porphyromonas gingivalis, Prevotella intermedia, and Tannerella forsythia (previously T. forsythensis) in patients with generalized AgP (AgP group), CP (CP group), or gingivitis (G group) and in healthy individuals (C group). METHODS: Subgingival plaque samples were collected with paper points from 30 patients in each group. The nested polymerase chain reaction (PCR) method was used to detect HSV-1, EBV-1, and HCMV. Bacteria were identified by 16S rRNA-based PCR. RESULTS: HSV-1, HCMV, and EBV-1 were detected in 86.7%, 46.7%, and 33.3% of the AgP group, respectively; in 40.0%, 50.0%, and 46.7% of the CP group, respectively; in 53.3%, 40.0%, and 20.0% of the G group, respectively; and in 20.0%, 56.7%, and 0.0% of the C group, respectively. A. actinomycetemcomitans was detected significantly more often in the AgP group compared to the other groups (P <0.005). P. gingivalis and T. forsythia were identified more frequently in AgP and CP groups, and AgP, CP, and G groups had higher frequencies of P. intermedia compared to the C group. CONCLUSION: In Brazilian patients, HSV-1 and EBV-1, rather than HCMV, were more frequently associated with CP and AgP.

Herpes viruses in periodontal compromised sites: comparison between HIV-positive and -negative patients.

AIM: The objective of this study was to compare the frequency of herpes simplex virus type 1 (HSV-1), Epstein-Barr virus (EBV) and human cytomegalovirus (HCMV) in subgingival plaque, saliva and peripheral blood of HIV-positive and-negative patients with periodontal disease. MATERIAL AND METHODS: Fifty HIV-positive subjects (23 with gingivitis, 27 with periodontitis) and 50 healthy HIV-negative patients with chronic periodontitis were included in the study. Parameters of probing depth (PD), clinical attachment level (CAL), gingival index and plaque index were recorded. The samples were processed for viral identification by the nested polymerase chain reaction technique. RESULTS: HCMV was the most prevalent virus in HIV-positive (82%) and-negative patients (84%), and the detection in the three samples was similar (p>0.05). HSV-1 was the least prevalent virus in both groups, being detected in similar frequencies in oral sites and in peripheral blood. EBV-1 was found more frequently in saliva and subgingival plaque of HIV-positive patients than in HIV-negative patients (p< or =0.05). CONCLUSIONS: EBV-1 was more frequently recovered in oral sites of HIV-positive patients than in HIV-negative patients.

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.

Detection of Epstein-Barr virus and human cytomegalovirus in blood and oral samples: comparison of three sampling methods.

The purpose of this study was to identify and compare the presence of HCMV and EBV-1 in subgingival plaque, unstimulated saliva and peripheral blood of patients with chronic periodontitis. Forty patients diagnosed with chronic periodontitis (mean age, 41.7 years) were recruited. Unstimulated saliva, subgingival plaque and peripheral blood were collected from each patient and the DNA of each sample was isolated. The viruses were detected using the nested PCR technique. The detection frequency of EBV-1 in subgingival plaque, saliva and peripheral blood was 45%, 37.5% and 25%, respectively. HCMV was detected in 82.5% of subgingival plaque samples and peripheral blood and in 75% of salivary samples. The sensitivity for detecting EBV-1 in saliva and peripheral blood when EBV-1 was detected in subgingival plaque samples was low (22% and 27.7%, respectively) and the sensitivity for detecting HCMV in saliva and peripheral blood when compared to subgingival plaque was high (81.8% and 87.8%, respectively). There is a high agreement among the three sampling methods in detection of HCMV, but the detection of EBV-1 would require a combination of saliva and subgingival plaque sampling to avoid false negative results.

Human herpesviruses and Porphyromonas gingivalis are associated with juvenile periodontitis.

BACKGROUND: Although herpesviruses have been associated with adult periodontitis, their relationship with juvenile periodontitis (JP) has not been established. This case-control study examined possible associations between JP and pathogenic bacteria, the human cytomegalovirus (HCMV), and the Epstein-Barr type 1 virus (EBV-1). METHODS: Subjects were participants in a larger survey of schoolchildren in North-Central Jamaica. Subgingival plaque samples from 15 subjects with JP, 20 with incipient periodontitis (IP), and 65 randomly-selected healthy controls were assayed for Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans using a 16S rRNA polymerase chain reaction (PCR) identification method, and for HCMV and EBV-1 using nested PCR identification. RESULTS: Strong bivariate associations were found between JP and P. gingivalis (odds ratio [OR] = 12.7; 95% CI = 2.6, 61.4), HCMV (OR = 10.0; 95% CI = 2.7, 36.3), and A. actinomycetemcomitans (OR = 8.0; 95% CI = 2.3, 27.5), but not EBV-1. In multivariate analyses, P. gingivalis remained a significant explanatory variable (OR = 7.8; 95% CI = 1.5, 40.9); however, the associations were marginal for HCMV (OR = 4.6; 95% CI = 0.9, 22.7), and non-significant for A. actinomycetemcomitans (OR = 2.0; 95% CI = 0.4, 9.7). The associations with JP and the extent of attachment loss were even stronger when both P. gingivalis and HCMV were detected together. P. gingivalis (OR = 3.9; 95% CI = 1.3, 12.0) and EBV-1 (OR = 3.3; 95% CI = 1.0, 10.3) were the only significant explanatory variables in the multivariate analysis of IP. CONCLUSIONS: P. gingivalis is the strongest and most stable indicator of periodontitis in Jamaican adolescents. Co-infection with P. gingivalis and HCMV appears to be particularly deleterious to periodontal health.