Aislamiento de bacterias periodontop ticas desde hemocultivos y ateromas obtenidos de pacientes con aterosclerosis y periodontitis / Isolation of periodontal bacteria from blood samples and atheromas in patients with atherosclerosis and periodontitis

Background: Periodontitis is a common oral disease produced by bacterial species that reside in the subgingival plaque. These microorganisms have been associated to atherosclerosis and it is suggested that periodontitis is a cardiovascular risk factor. Aim: To isolate periodontal bacteria from blood and atheroma samples, from patients with atherosclerosis and periodontitis. Material and methods: Twelve patients with periodontitis and a clinical diagnosis of atherosclerosis and 12 patients with periodontitis but without atherosclerosis were studied. Blood samples were obtained immediately before and after scaling and root planing. The samples were incubated in aerobic and anaerobic conditions. One week after scaling, atheromatous plaques were obtained during endarterectomy in the 12 patients with atherosclerosis. These were homogenized and cultured for aerobic and anaerobic bacteria. Microorganisms were identified by means ofPCR. Results: Five patients with and two without atherosclerosis, had bacteremia after scaling and root planing. Bacterial species isolated from blood samples were the same found in periodontic pockets. Four atheromatous plaques of patients with bacteremia yielded positive cultures. The isolated bacteria were the same found in blood samples and periodontal pockets. Conclusions: Bacteremia occurred in seven of 24 patients after scaling and root planing. In four patients, the same species found in periodontic pockets and blood cultures were detected in atherosclerotic plaques obtained one week after the dental procedure.

Chlamydia pneumoniae e Mycoplasma pneumoniae nos nódulos de calcificacão da estenose da valva aórtica / Chlamydia pneumoniae and Mycoplasma pneumoniae in calcified nodes of stenosed aortic valves

OBJETIVO: Investigar se chlamydia pneumoniae (CP) ou mycoplasma pneumoniae (MP) estão presentes na estenose da valva aórtica (EA). MÉTODOS: Imuno-histoquímica foi utilizada para identificar os antígenos de CP (Ag-CP), a hibridizacão in situ para identificar o DNA de MP, e microscopia eletrônica para avaliacão dos dois agentes, nos grupos: normal - 11 valvas normais de autópsia; aterosclerose - 10 valvas de pacientes com aterosclerose sistêmica de autópsia e sem EA; e EA - 14 espécimes cirúrgicos provenientes de pacientes com EA analisados em 3 sub-regiões: EA-preservada - regiões mais preservadas na periferia da valva; EA-fibrose - tecido fibrótico peri-calcificacão; e EA-calcificacão - nódulos calcificados. RESULTADOS: As medianas da fracão de área positiva para Ag-CP foram 0,09; 0,30; 0,18; 1,33; e 3,3 nos grupos acima descritos, respectivamente. A densidade de CP foi significativamente maior nos grupos aterosclerose e EA-calcificacão em relacão ao normal (p<0,05). Dentro do grupo EA, a quantidade de CP foi maior nas regiões de fibrose e calcificacão (p<0,05). As fracões de área positivas para MP-DNA (medianas) foram 0,12; 0,44; 0,07; 0,36; e 1,52 nos grupos acima descritos, respectivamente. A quantidade de MP-DNA foi maior na EA-calcificacão em relacão ao normal (p<0,05). Dentro do grupo EA, maior quantidade de MP-DNA foi encontrada nas regiões de calcificacão e fibrose (p<0,05). CONCLUSAO: Os nódulos de calcificacão da EA tinham maior concentracão de CP e MP sugerindo que essas bactérias possam estar associadas ao desenvolvimento de calcificacão e inflamacão, apontando novas semelhancas entre os processos de EA e aterosclerose, que podem ter mecanismos infecciosos envolvidos.

Efecto de la infección periodontal por porphyromonas gingivalis en el desarrollo de ateroesclerosis en ratones deficientes de apolipoproteína E / Effects of porphyromona gingivalis periodontal infection upon atherosclerosis development in apolipoprotein-E

Introducción: Nuestro grupo recientemente demostró una asociación significativa entre periodontitis, placas coronarias aguda y extensión de la enfermedad coronaria aterosclerótica en pacientes con síndrome coronario agudo. Objetivo: Desarrollar un modelo experimental animal para estudiar el posible efecto pro-aterogénico de la inducción de periodontitis por Porphyromona Gingivalis (PG) en ratones deficientes en la apolipoproteína E (APO-E KO). Métodos: En 12 ratones APO-E KO mantenidos con dieta hiperlipidémica se realizaron tocaciones con PG cepa ATCC 53977 en el surco gingival de los molares mandibulares a las 8 semanas de vida. Igual número de ratones APO-E KO fue intervenido con el mismo procedimiento, pero sólo con el vehículo de las tocaciones. Estos procedimientos se repitieron a las 48, 72 y 120 hrs de la infección inicial. Luego de 4 semanas post-inoculación con PG se realizaron estudios histomorfométricos en la aorta proximal para medir la severidad de las lesiones ateromatosas y en las mandíbulas, para evaluar la pérdida del hueso alveolar. Resultados: No se observó una diferencia significativa en el daño del hueso alveolar en las mandíbulas de los animales infectados versus el grupo control. En las aortas, la razón tamaño placa/pared vascular fue mayor en el grupo infectado con PG que en el grupo control (0.132 ± 0.2 versus 0.103 ± 0.15, respectivamente), pero esta diferencia no fue estadísticamente significativa. Conclusión: El diseño experimental del presente estudio no permitió establecer si la periodontitis inducida por PG es capaz o no de acelerar el proceso aterogénico de los ratones APO-E KO. Será necesario aplicar un protocolo de infección periodontal más agresivo en estos animales para evaluar más adecuadamente el efecto de PG sobre la ateroesclerosis.

Chlamydia pneumoniae infection and nonspecific aortoarteritis: search for a link with a nonatherosclerotic inflammatory arterial disease.

BACKGROUND: The association between Chlamydia pneumoniae infection and atherosclerosis has gained recognition. However, the nature of this association is controversial. The infective link may not be specific for atherosclerosis and may also exist in other nonatherosclerotic arterial diseases. We investigated patients with nonspecific aortoarteritis for serological evidence of prior Chlamydia pneumoniae infection. METHODS AND RESULTS: Fifty patients each of nonspecific aortoarteritis and coronary artery disease with angiographic evidence of significant (>70%) coronary artery lesions were tested for the presence of IgG antibodies against Chlamydia pneumoniae by micro-immunofluorescence assay and compared with 50 age- and sex-matched normal healthy controls. The number of patients with nonspecific aortoarteritis who tested positive for Chlamydia pneumoniae antibodies (IgG) was not significantly different from controls (8 v. 7, p=ns). The mean titer amongst positive subjects in the two groups was also similar (1:40+/-40 v. 1:50+/-25; p=ns). Patients with coronary artery disease were significantly older than patients with nonspecific aortoarteritis and controls (53.2+/-5.8 v. 21.2+/-9.9 years and 24.5+/-5.2 years, p<0.01 for both) and showed a higher seroprevalence of prior Chlamydia pneumoniae infection (18 v. 8 and 7, p < 0.05 for both). The mean IgG titers of patients with coronary artery disease who tested positive were also significantly higher than the other two groups (1:98+/-34 v. 1:40+/-40, p<0.001 and 1:98+/-34 v. 1:50+/-25, p<0.01, respectively). CONCLUSIONS: In patients with nonspecific aortoarteritis, the seroprevalence of prior Chlamydia pneumnoniae infection is not more than that in healthy individuals of the same age group, but is significantly lesser than that in patients with coronary artery disease. Thus Chlamydia pneumoniae infection may not be associated with all forms of chronic inflammatory arterial lesions.

Serologic and histopathologic study of Chlamydia pneumoniae infection in atherosclerosis: a possible pathogenetic mechanism of atherosclerosis induced by Chlamydia pneumoniae

Chronic infection and inflammation have recently been implicated as important etiologic agents for atherosclerosis in general and, in particular, ischemic heart disease. Several agents have been suggested as possible candidates for the chronic inflammation including cytomegalovirus, Helicobacter pylori and Chlamydia pneumoniae. We hypothesized that a vascular infection with C. pneumoniae may induce a chronic inflammatory reaction in the host vascular tissue and activated inflammatory cells may express inflammatory mediators such as cyclooxygenase-2 (COX-2) and matrix metalloproteinases (MMPs). At first, we evaluated the relationship between C. pneumoniae infection and atherosclerosis indirectly by serologic study, and then, to confirm our hypothesis, we performed an immunohistochemical study of atherosclerotic plaques. The seropositive rate of anti-Chlamydia pneumoniae IgG was higher in the disease group (Group I, 59.8%, n = 254) than in the negative control group (Group III, 47.4%, n = 97) (p = 0.041), but the anti-Chlamydia pneumoniae IgA was not different in seropositivity between the two groups (Group I, 64.6%; Group III, 57.7%). The simultaneous seropositive rates of both IgG and IgA were 56.7% in Group I and 43.3% in Group III (p = 0.033). In subgroups without the conventional risk factors of atherosclerosis, these findings were more prominent. Furthermore, we performed immunohistochemical staining on the atherosclerotic aortic tissues obtained from patients that were seropositive to C. pneumoniae (n = 5), by using antibodies to C. pneumoniae, COX-2, and MMP-9. The immunoreactivity for COX-2 and MMP-9 increased in the atherosclerotic plaques itself, predominantly in the surrounding area of immunoreactive C. pneumoniae. These findings support our hypothesis and C. pneumoniae may participate in a pathogenetic mechanism for atherogenesis or progression of atherosclerosis. The present study may open a promising perspective concerning future therapeutic trials of chronic inflammation related atherogenesis under pathophysiological conditions.