RESUMO
INTRODUCTION: We recently investigated global gene expression in ST2 mouse stromal cells infected by the periodontal pathogen Porphyromonas gingivalis using microarray technology, and found that the bacterium induces a wide range of proinflammatory gene expression. Here, we reported the signaling pathways involved in those proinflammatory responses. METHODS: ST2 cells and primary calvarial osteoblasts from C3H/HeN, C57BL/6, and MyD88-deficient (MyD88(-/-)) mice were infected with P. gingivalis ATCC33277 and its gingipain-deficient mutant KDP136. Expression of the chemokines CCL5 and CXCL10, and matrix metalloproteinase-9 (MMP9) were quantified by real-time polymerase chain reaction, while phosphorylation of protein kinases and degradation of an inhibitor of nuclear factor-kappaB, IkappaB-alpha, were detected by Western blotting, and activation of transcriptional factors was determined by a luciferase reporter assay. The effects of inhibitors of transcriptional factors and protein kinases were also investigated. RESULTS: Infection by P. gingivalis elicited gene expression of CCL5, CXCL10, and MMP9 in both ST2 cells and osteoblasts. Western blot and reporter assay results revealed activation of nuclear factor-kappaB (NF-kappaB) and activator protein-1 transcription factors. The NF-kappaB inhibitor suppressed the expression of CCL5 and MMP9, but not that of CXCL10, whereas P. gingivalis infection induced significant CCL5 expression in MyD88(-/-) osteoblasts. In addition, activation of protease-activated receptors by trypsin elicited significant induction of CXCL10. CONCLUSION: Our results suggest that various proinflammatory responses in P. gingivalis-infected stromal/osteoblast cells are NF-kappaB-dependent, but not always dependent on the Toll-like receptor/MyD88 pathway, while some responses are related to the activation of protease-activated receptors. Thus, P. gingivalis does not fully utilize well-established pathogen recognition molecules such as Toll-like receptors.
Assuntos
Quimiocina CCL5/biossíntese , Quimiocina CXCL10/biossíntese , Inflamação/metabolismo , Metaloproteinase 9 da Matriz/biossíntese , NF-kappa B/metabolismo , Osteoblastos/metabolismo , Osteoblastos/microbiologia , Porphyromonas gingivalis/fisiologia , Transdução de Sinais , Animais , Western Blotting , Células da Medula Óssea/metabolismo , Células da Medula Óssea/microbiologia , Linhagem Celular , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fator 88 de Diferenciação Mieloide/metabolismo , Células Estromais/metabolismo , Células Estromais/microbiologia , Fator de Transcrição AP-1/metabolismoRESUMO
OBJECTIVES: We investigated the relationship between phenytoin-induced gingival overgrowth (GO) and the harboring of periodontal bacteria. MATERIALS AND METHODS: Periodontal conditions and subgingival bacterial profiles were examined in 450 sites of 75 subjects. A polymerase chain reaction method was used to detect six bacterial species; Porphyromonas gingivalis (Pg), Actinobacillus actinomycetemcomitans (Aa), Tannerella forsythia, Treponema denticola (Td), Prevotella intermedia (Pi), and Prevotella nigrescens (Pn). Genetic variations of the Pg fimA gene were also examined. Bacterial occurrence was compared with the severity of GO, and alterations in the bacterial occurrence rate and quantities were monitored following periodontal treatment. RESULTS: The occurrences of Aa, Td, Pi, Pn, and Pg with type II fimA (type II Pg) were significantly associated with the severity of GO. Td occurrence was reduced in association with gingival improvement following ultrasonic scaling, however, no such relationship was observed with Aa, Pi, Pn, and Pg. In addition, Pg and Pi markedly persisted after treatment. Clinical improvement of the sites, following an Er:YAG laser treatment, significantly associated with quantitative reduction of Pg in improved sites, however, not that of Pi. CONCLUSION: Type II Pg and Td were each found to have a significant relationship with the development and deterioration of GO.
Assuntos
Proteínas de Fímbrias/genética , Crescimento Excessivo da Gengiva/microbiologia , Porphyromonas gingivalis/química , Porphyromonas gingivalis/patogenicidade , Treponema denticola/patogenicidade , Adolescente , Adulto , Aggregatibacter actinomycetemcomitans/patogenicidade , Anticonvulsivantes/efeitos adversos , Bacteroides/patogenicidade , Criança , Placa Dentária/microbiologia , Raspagem Dentária , Feminino , Variação Genética , Crescimento Excessivo da Gengiva/induzido quimicamente , Crescimento Excessivo da Gengiva/terapia , Humanos , Terapia a Laser , Masculino , Fenitoína/efeitos adversos , Reação em Cadeia da Polimerase , Porphyromonas gingivalis/genética , Prevotella/patogenicidadeRESUMO
Porphyromonas gingivalis fimbriae are critical for the promotion of bacterial infection. The fimA gene encoding fimbrillin, a subunit of fimbriae, has been classified into five genotypes (types I to V) based on their nucleotide sequences. Using a fimA type-specific PCR assay, our previous study demonstrated a close relationship between P. gingivalis possessing type II and type IV fimA genes and adult periodontitis. In that study, some clinical specimens were found to be positive for both types I- and II- fimA specific primers, likely due to the coexistence of two clonal types or a single clone of an unknown genotype in the samples. In the present study, we cloned a new variant of the fimA gene, designated as type Ib fimA, from P. gingivalis HG1691. The nucleotide sequence of the cloned fimA gene showed a 97.1% homology with that of type I fimA, indicating it as a clonal variant of type I fimA. Organisms with type Ib fimA were detected in 13.5% of periodontitis patients and in 2.9% of periodontal healthy adults. Statistical analysis revealed a strong relationship between periodontitis and specific fimA types such as type Ib [odds ratio (OR) 6.51], type II (OR 77.8), and type IV (OR 7.54). Moreover, type Ib fimA-organisms were also found to be related to periodontitis in Down's syndrome (OR 1.91) and mentally disabled populations (OR 4.00). These findings suggest that P. gingivalis with type Ib fimA is closely associated with the progression of periodontitis, similar to organisms with type II and IV fimA.
Assuntos
Proteínas de Fímbrias/genética , Periodontite/microbiologia , Pessoas com Deficiência Mental , Pili Sexual/genética , Porphyromonas gingivalis/genética , Adulto , Idoso , Sequência de Bases , Estudos de Casos e Controles , Células Clonais , Intervalos de Confiança , Placa Dentária/microbiologia , Síndrome de Down/microbiologia , Feminino , Proteínas de Fímbrias/classificação , Genótipo , Humanos , Deficiência Intelectual/microbiologia , Masculino , Pessoa de Meia-Idade , Razão de Chances , Pili Sexual/classificação , Porphyromonas gingivalis/classificação , Homologia de Sequência do Ácido NucleicoRESUMO
The purpose of this study was to examine characteristic profiles of Streptococcus pyogenes clinical isolates isolated in Japan during 1994-9. Genotyping of the M protein (emm typing) revealed that emm types 12 and 28 were the most common among 316 isolates. Most of the emm12 isolates were isolated from mucosa, while emm58 and emm89 were from skin. Moreover, the emm3 isolates were dominant in invasive infections. The distribution of 6 superantigen genes showed that all isolates harboured the mf gene and many had the speG gene. Invasive isolates were shown to have the ssa gene at a higher rate (76%) than noninvasive (37%). The distribution of superantigens was significantly different between emm types, but not between isolation sites. These results suggest that the distribution of emm types is related to isolation site, whereas superantigen distribution is related to clinical features of S. pyogenes infections.
Assuntos
Antígenos de Bactérias , Proteínas da Membrana Bacteriana Externa/genética , Proteínas de Transporte/genética , Infecções Estreptocócicas/patologia , Streptococcus pyogenes/genética , Streptococcus pyogenes/patogenicidade , Superantígenos/análise , Genótipo , Humanos , Reação em Cadeia da Polimerase , Infecções Estreptocócicas/genética , Streptococcus pyogenes/imunologiaRESUMO
A case history of a woman with gingival overgrowth (GO) induced by amlodipine is presented. A 49-year-old Japanese woman, who was taking amlodipine, had gingival overgrowth and swelling on examination. No specific periodontal treatment was provided to the patient for the GO; however, the amlodipine was replaced with an ACE inhibitor after consultation with her medical practitioner. Within two months, the suspension of amlodipine resulted in a significant improvement in her periodontal condition. Failure to control the hypertension caused the physician to re-prescribe amlodipine. After three months, the gingival overgrowth returned; however, its severity was less when compared with the original periodontal condition, due to reduction in drug dose and periodontal therapy. This experience suggests that temporary suspension of a drug which can induce GO can improve the periodontal condition without the aid of surgical treatment.
Assuntos
Anlodipino/efeitos adversos , Anti-Hipertensivos/efeitos adversos , Bloqueadores dos Canais de Cálcio/efeitos adversos , Crescimento Excessivo da Gengiva/induzido quimicamente , Antagonistas Adrenérgicos beta/uso terapêutico , Anlodipino/administração & dosagem , Inibidores da Enzima Conversora de Angiotensina/uso terapêutico , Anti-Hipertensivos/administração & dosagem , Anti-Hipertensivos/uso terapêutico , Bloqueadores dos Canais de Cálcio/administração & dosagem , Feminino , Hemorragia Gengival/induzido quimicamente , Hemorragia Gengival/prevenção & controle , Crescimento Excessivo da Gengiva/prevenção & controle , Bolsa Gengival/induzido quimicamente , Bolsa Gengival/prevenção & controle , Humanos , Pessoa de Meia-Idade , RecidivaRESUMO
BACKGROUND: Down's syndrome (DS) patients often develop severe early-onset marginal periodontitis in early adulthood; however, there is little information available on the microbiology of DS periodontitis. METHODS: Subgingival plaque specimens were taken from 67 DS young adults and 41 age-matched systemically healthy individuals with mental disabilities (MD). The prevalence of 10 possible periodontopathic bacterial species, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Bacteroides forsythus, Treponema denticola, Prevotella intermedia, Prevotella nigrescens, Capnocytophaga ochracea, Capnocytophaga sputigena, Campylobacter rectus, and Eikenella corrodens, were investigated in their subgingival plaque samples using a polymerase chain reaction method. The detection of P. gingivalis fimA genotypes was also performed in P. gingivalis-positive samples. RESULTS: Although DS subjects generally develop an earlier and more extensive periodontal breakdown than those with MD, no significant differences were observed in the bacterial profiles. The profiles of subjects with periodontitis were significant in DS, but not in MD. The prevalence of P. gingivalis, B. forsythus, and P. intermedia were significant in the DS periodontitis group, compared to DS gingivitis group. Moreover, the occurrence of P. gingivalis with the type II fimA gene was significantly related to periodontitis in both DS and MD, with odds ratios of 6.32 and 12.03, respectively. CONCLUSIONS: These results suggest that early-onset periodontitis in DS is mainly due to the more susceptible host for the causative microbial agents including P. gingivalis with type II fimA.
Assuntos
Periodontite Agressiva/microbiologia , Bactérias/classificação , Síndrome de Down/microbiologia , Proteínas de Fímbrias , Adulto , Aggregatibacter actinomycetemcomitans/crescimento & desenvolvimento , Proteínas de Bactérias/genética , Bacteroides/crescimento & desenvolvimento , Campylobacter/crescimento & desenvolvimento , Capnocytophaga/crescimento & desenvolvimento , Estudos de Casos e Controles , Intervalos de Confiança , Placa Dentária/microbiologia , Eikenella corrodens/crescimento & desenvolvimento , Feminino , Genótipo , Humanos , Masculino , Transtornos Mentais/microbiologia , Razão de Chances , Pili Sexual/genética , Porphyromonas gingivalis/genética , Porphyromonas gingivalis/crescimento & desenvolvimento , Prevalência , Prevotella/crescimento & desenvolvimento , Prevotella intermedia/crescimento & desenvolvimento , Estatística como Assunto , Treponema/crescimento & desenvolvimentoRESUMO
Periodontopathogenic bacteria survive various environmental changes during the progression of periodontal disease. Alterations in metabolism and protein expression will have to take place to adapt their physiological functions to environmental stress. We examined the effects of an elevation of 2 degrees C in temperature on the adhesive ability and antigenicity of Porphyromonas gingivalis. Elevation of growth temperature of P. gingivalis from 37 degrees C to 39 degrees C remarkably suppressed the expression of surface filamentous structures, such as fimbriae, as well as the adhesive capacities to salivary components and Streptococcus oralis. Sera of severe periodontitis patients revealed a marked increase in serological activity with 39 degrees C cells than with 37 degrees C cells. The alteration of protein profiles of bacterial surface components by temperature elevation was demonstrated by SDS-PAGE, and their Western blot profiles were also different from those of cells grown at 37 degrees C. Although a uniform trend was not found in the altered patterns, sera from severe periodontitis patients detected more antigenic proteins in cells grown at 39 degrees C than 37 degrees C cells. These observations suggest that P. gingivalis downregulates the expression of fimbriae and alters its adhesive capacity and antigenicity by the temperature stress that could occur during the disease progression.
Assuntos
Aderência Bacteriana/fisiologia , Periodontite/microbiologia , Porphyromonas gingivalis/fisiologia , Adaptação Fisiológica , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/imunologia , Regulação para Baixo , Ensaio de Imunoadsorção Enzimática , Fímbrias Bacterianas/fisiologia , Temperatura Alta , Humanos , Periodontite/imunologia , Porphyromonas gingivalis/imunologia , Estresse FisiológicoRESUMO
The effects of nifedipine on the death and proliferation of gingival fibroblasts were investigated to elucidate the mechanism of gingival overgrowth that is associated with chronic administration of Ca2+ channel blockers. The number of adhered viable and dead fibroblasts obtained from healthy human gingiva increased after confluence, whereas cell death was inhibited by nifedipine in a concentration-dependent manner. A similar inhibition was also observed in the presence of other calcium channel blockers, such as nicardipine, diltiazem, and verapamil. When gingival fibroblasts were co-cultured with RAW264 (macrophage-like) cells, lipopolysaccharide (LPS) caused the concentration-dependent death of fibroblasts. Nifedipine significantly inhibited the LPS-induced cell death. Although neither LPS nor N-ethyl-2-(1-ethyl-2-hydroxy-2-nitroso-hydrazino)-ethanamine, a nitric oxide donor, directly caused fibroblast death, 3-morpholino-sydnonimine (SIN-1), a peroxynitrite donor, induced fibroblast death, regardless of the presence of RAW cells. The cell death induced by SIN-1 was not affected by nifedipine treatment. LPS stimulation caused an increase in the immunoreactivity of inducible nitric oxide synthase (iNOS) and in the nitrite concentration in the incubation medium of RAW cells. The induction of iNOS was completely prevented by the incubation with nifedipine. The inhibition by nifedipine of nitrite production in RAW cells was also observed after treatment with nicardipine, but not with either diltiazem or verapamil. Therefore, the inhibition by nifedipine of both adherence- and LPS-stimulated macrophage-induced death of fibroblasts may be the mechanism of gingival overgrowth seen during chronic treatment with Ca(2+) channel blockers.
Assuntos
Bloqueadores dos Canais de Cálcio/farmacologia , Adesão Celular/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Macrófagos/fisiologia , Nifedipino/farmacologia , Animais , Divisão Celular/efeitos dos fármacos , Linhagem Celular , Células Cultivadas , Técnicas de Cocultura , Fragmentação do DNA/efeitos dos fármacos , Diltiazem/farmacologia , Relação Dose-Resposta a Droga , Fibroblastos/citologia , Fibroblastos/metabolismo , Gengiva/citologia , Gengiva/efeitos dos fármacos , Humanos , Lipopolissacarídeos/farmacologia , Macrófagos/citologia , Nicardipino/farmacologia , Óxido Nítrico/metabolismo , Doadores de Óxido Nítrico/farmacologia , Timidina/metabolismo , Fatores de Tempo , Verapamil/farmacologiaRESUMO
Porphyromonas gingivalis fimA gene encoding fimbrillin, a subunit of fimbriae, has been classified into 5 genotypes (types I to V) based on their nucleotide sequences. Here, we investigated the relationship between the prevalence of these fimA genotypes and periodontal health status in adults. Dental plaque specimens obtained from 380 periodontally healthy adults and 139 periodontitis patients were analyzed by the PCR method. P. gingivalis was detected in 36.8% of the healthy subjects and in 87.1% of the periodontitis patients. Among the P. gingivalis-positive healthy adults, the most prevalent fimA type was type I (76.1%), followed by type V. In contrast, a majority of the periodontitis patients carried type II fimA organisms (66.1%), followed by type IV. The univariate analysis illustrated that periodontitis was associated with the occurrences of type I fimA (OR 0.16), type II (OR 44.44), type III (1.96), type IV (13.87), and type V (1.40). These findings clearly indicate that there are both disease-associated and non-disease-associated strains of P. gingivalis, and that their infectious traits influencing periodontal health status could be differentiated based on the clonal variation of fimA genes.
Assuntos
Proteínas de Bactérias/genética , Proteínas de Fímbrias , Periodonto/microbiologia , Pili Sexual/genética , Porphyromonas gingivalis/genética , Adulto , Idoso , Sequência de Bases , Distribuição de Qui-Quadrado , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Periodontite/epidemiologia , Periodontite/microbiologia , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/estatística & dados numéricos , Porphyromonas gingivalis/isolamento & purificação , PrevalênciaRESUMO
A case of familial hypophosphatemic vitamin D-resistant rickets or X-linked hypophosphatemia (XLH) accompanied by specific systemic and dental findings is reported. A 15-year-old boy with XLH visited our facility complaining of a toothache in the left lower canine region. Two other family members of the patient, his younger sister and their mother, also had XLH, whereas the other 2 members, his younger brother and father, are healthy. Those with XLH show systemic signs of the disease, such as growth retardation, limb deformity, and spinal curvature disorders; however, these symptoms are more severe in the patient than in the others. The patient had multiple periodontal abscesses, but no evidence of dental caries, trauma, or periodontal disease on the corresponding teeth at the time of his oral examination. A radiographic examination showed root dysplasia and enlarged pulp chambers.A histologic examination of an extracted third molar showed marked globular dentin and an increased predentin width. The abscess was thought to be caused by pulpal infection, which came from bacterial invasion through enamel cracks and dentinal microcleavage of the teeth. The treatments provided in this case are discussed.
Assuntos
Hipofosfatemia Familiar/complicações , Dente Serotino/anormalidades , Anormalidades Dentárias/etiologia , Adolescente , Dente Canino , Esmalte Dentário/anormalidades , Dentina/anormalidades , Humanos , Hipofosfatemia Familiar/genética , Hipofosfatemia Familiar/patologia , Masculino , Mandíbula , Dente Serotino/patologia , Linhagem , Abscesso Periapical/complicações , Abscesso Periapical/etiologia , Abscesso Periapical/terapia , Radiografia Panorâmica , Tratamento do Canal Radicular , Anormalidades Dentárias/complicações , Anormalidades Dentárias/patologiaRESUMO
BACKGROUND: Cyclosporine A (CsA) and calcium channel blockers induce gingival overgrowth in humans and animals. Recently, nifedipine and diltiazem have often been used to control CsA-related hypertension in organ transplant patients. The purpose of this study was to examine the effects of a combined oral treatment of CsA and nifedipine or diltiazem on the severity of gingival overgrowth in rats. METHODS: Fifteen-day-old Fischer rats were treated orally with single or combined applications of CsA, nifedipine, and/or diltiazem for 40 days; and induced gingival overgrowth, rat growth, and blood drug levels were compared among the different experimental groups. The experiment consisted of 6 groups: one control group (group A) and 5 test groups treated with CsA (group B), nifedipine (group C), and diltiazem (group D), as well as those concurrently treated with CsA and nifedipine (group E), and CsA and diltiazem (group F). Gingival overgrowth was determined by measuring the depth of the gingival sulcus. RESULTS: The mandibular buccal gingival sulcus depth of group A was 365 +/- 41.2 microm. Among the test groups, the most remarkable gingival overgrowth was seen in group E (1,020 +/- 63.3 microm), followed by group F (895 +/- 43.8 microm), group B (870 +/- 48.3 microm), group C (525 +/- 116 microm), and then group D (505 +/- 83.2 microm). Rat body weight gain was reduced significantly by oral CsA treatment. Neither nifedipine nor diltiazem suppressed rat growth when used independently; however, rat growth reduced by CsA was further suppressed by a combined use of diltiazem, but not nifedipine. CsA blood levels were reduced by concurrent oral treatment with nifedipine or diltiazem along with the blood levels of those calcium channel blockers when treatment was in combination with CsA. CONCLUSIONS: These results suggest that gingival overgrowth is induced in rats as a side effect of CsA, nifedipine, or diltiazem, and the combined use of these drugs influences rat growth, blood drug levels, and the severity of gingival overgrowth.
Assuntos
Bloqueadores dos Canais de Cálcio/efeitos adversos , Ciclosporina/efeitos adversos , Diltiazem/efeitos adversos , Crescimento Excessivo da Gengiva/induzido quimicamente , Imunossupressores/efeitos adversos , Nifedipino/efeitos adversos , Administração Oral , Análise de Variância , Animais , Peso Corporal/efeitos dos fármacos , Bloqueadores dos Canais de Cálcio/administração & dosagem , Bloqueadores dos Canais de Cálcio/sangue , Ciclosporina/administração & dosagem , Ciclosporina/sangue , Diltiazem/administração & dosagem , Diltiazem/sangue , Combinação de Medicamentos , Interações Medicamentosas , Gengiva/efeitos dos fármacos , Crescimento Excessivo da Gengiva/classificação , Crescimento/efeitos dos fármacos , Imunossupressores/administração & dosagem , Imunossupressores/sangue , Masculino , Nifedipino/administração & dosagem , Nifedipino/sangue , Ratos , Ratos Endogâmicos F344 , Método Simples-CegoRESUMO
BACKGROUND: It is widely known that individuals with Down syndrome (DS) often develop severe early-onset periodontal diseases. In this study, we examined the prevalence of periodontopathic bacteria in DS children to determine if specific pathogens are acquired in their childhood. METHODS: The subjects were 60 DS children (2 to 13 years old, 5 in each age bracket) and 60 age-matched controls. Ten pathogens, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Bacteroides forsythus, Treponema denticola, Prevotella intermedia, P nigrescens, Capnocytophaga ochracea, C. sputigena, Campyrobacter rectus, and Eikenella corrodens were surveyed in subgingival plaque samples using a polymerase chain reaction. Periodontal status was evaluated by probing depth, bleeding on probing, and gingival index. RESULTS: No significant difference in periodontal status was observed between the DS and control groups, however, all of the pathogens were detected with greater frequency in the DS children. B. forsythus, T. denticola, P. nigrescens, and C. rectus were significantly prevalent throughout all age brackets of the DS children (P <0.01 or 0.05). The occurrence of P. gingivalis was also significant in the DS subjects over 5 years old. A cluster analysis of the microbial profiles of the DS subjects showed that gingivitis severity was associated with increased varieties of the harboring pathogens and the distribution of P. gingivalis. CONCLUSIONS: These results suggest that various periodontopathogens can colonize in the very early childhood of DS patients and maturation of subgingival components, including P. gingivalis, plays an important role in the initiation of gingival inflammation.
Assuntos
Assistência Odontológica para Doentes Crônicos , Síndrome de Down/microbiologia , Doenças Periodontais/microbiologia , Adolescente , Aggregatibacter actinomycetemcomitans/isolamento & purificação , Bacteroides/isolamento & purificação , Campylobacter/isolamento & purificação , Capnocytophaga/isolamento & purificação , Estudos de Casos e Controles , Criança , Pré-Escolar , Análise por Conglomerados , Contagem de Colônia Microbiana , DNA Bacteriano/análise , Placa Dentária/microbiologia , Eikenella corrodens/isolamento & purificação , Feminino , Humanos , Masculino , Reação em Cadeia da Polimerase , Porphyromonas gingivalis/isolamento & purificação , Prevotella/isolamento & purificação , Treponema/isolamento & purificaçãoRESUMO
Cyclosporin A (CsA) is used as an immunosuppressive agent and its prominent side effect is the induction of fibrous gingival overgrowth. The purpose of this study was to investigate the effect of CsA on the type I collagen metabolism in the gingiva of rats fed a powdered diet either containing or lacking CsA. Immunohistochemical analysis revealed that type I collagen was more prevalent in the connective tissue of CsA-treated gingiva than in those of control rats on days 15, 30, and 55 after the start of feeding. Total RNAs were isolated from mandibular molar gingiva on days 0, 3, 8, 15, 30, and 55. Quantitative analysis of mRNA by reverse transcriptase-polymerase chain reaction revealed that the CsA-treated groups showed a gradual decrease in expression of type I collagen and collagenase mRNAs, 0.4% and 18.0% on day 55 compared with those on day 0, respectively. In the control groups, type I collagen and collagenase mRNAs also decreased to 19.7% and 63.0%, respectively, however, both mRNA expressions were significantly lower in the CsA-treated group than in the controls. An electron microscopic analysis of fibroblasts was performed to count the number of cells with collagen fibrils in the cytoplasm, a marker of phagocytosis of collagen by fibroblasts. The collagen fibrils were detected in 4.7% +/- 2.7% and 24.3% +/- 13.7% of fibroblasts in the overgrown gingiva treated with CsA rat for 8 days and 30 days, but in 57.0% +/- 5.3% and 81.3% +/- 9.2% of fibroblasts in the each control group gingiva, respectively. Furthermore, in vitro analysis was performed to measure the phagocytosis of cultured fibroblasts by flow cytometry using collagen-coated latex beads. Fibroblasts isolated from CsA-treated gingiva on day 8 and day 30 contained 5.7% +/- 0.6% and 9.9% +/- 1.5% phagocytic cells, whereas control fibroblasts contained 50.3% +/- 5.5% and 33.3% +/- 4.9% phagocytic cells, respectively. The inhibition rate of phagocytic activity was similar between in vivo and in vitro assays. These findings suggest that the decrease of the collagen degradation due to the lower phagocytosis and the lower collagenase mRNA expression are closely associated with the increase of type I collagen accumulation in CsA-treated rat gingiva.
Assuntos
Colágeno/metabolismo , Ciclosporina/efeitos adversos , Crescimento Excessivo da Gengiva/induzido quimicamente , Imunossupressores/efeitos adversos , Animais , Divisão Celular/efeitos dos fármacos , Células Cultivadas , Colágeno/genética , Colágeno/ultraestrutura , Colagenases/metabolismo , Fibroblastos/efeitos dos fármacos , Fibroblastos/enzimologia , Fibroblastos/ultraestrutura , Expressão Gênica/efeitos dos fármacos , Gengiva/química , Gengiva/efeitos dos fármacos , Gengiva/patologia , Crescimento Excessivo da Gengiva/metabolismo , Crescimento Excessivo da Gengiva/patologia , Látex , Masculino , Microscopia Eletrônica , Microesferas , Fagocitose/fisiologia , RNA Mensageiro/análise , Ratos , Ratos Endogâmicos F344 , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Organismos Livres de Patógenos EspecíficosRESUMO
OBJECTIVE: For the purpose of supplementing the shortage of dental information about DiGeorge syndrome, we report two cases of the syndrome seen in Japanese boys. STUDY DESIGN: Two cases were compared with respect to orofacial and dental findings; one was a case of complete DiGeorge syndrome and the other a case of partial DiGeorge syndrome. Extracted deciduous teeth from the two boys underwent histologic study. RESULTS: Each patient showed systemic developmental delay, hypocalcemia, and slight mental retardation. In the orofacial area, hypertelorism, a short philtrum, thick and reflected lips, and hypoplasia of the nasopharynx were also observed. A dental examination showed delayed formation and eruption of permanent teeth, aplasia of the nasopharynx, and enamel hypoplasia along with enamel hypocalcification. Structural streaks with increased calcification were histologically detected in the deciduous tooth from the patient with complete DiGeorge syndrome. CONCLUSIONS: Common characteristic orofacial and dental findings were noted in the two DiGeorge syndrome cases. Furthermore, histologic study of the deciduous tooth from the boy with complete DiGeorge syndrome suggests that there was some relationship between transient relative hypercalcemia and dentinal hypermineralized streaking of the tooth.
Assuntos
Hipoplasia do Esmalte Dentário/etiologia , Síndrome de DiGeorge/complicações , Síndrome de DiGeorge/patologia , Dente Decíduo/patologia , Adolescente , Pré-Escolar , Fácies , Humanos , MasculinoRESUMO
We evaluated the usefulness of a short professional mechanical tooth-cleaning (PMTC) program to improve periodontal conditions and caries susceptibility in 10 young adult patients with mental and/or physical disabilities. The PMTC program was carried out once on each of 6 sextants of the full mouth during 6 visits at two-week intervals. Even one treatment with PMTC was found to be significantly effective in reducing the probing depth in eight of the 10 subjects. A reduction in the total number of bleeding sites on probing was also clearly observed in all subjects. Moreover, the debris index was reduced in nine subjects by the PMTC program. Although caries susceptibility was improved, albeit very slowly, by PMTC, the Cariostat pH values showed no consistent tendency. The effects lasted for more than 6 weeks. Analysis of these results suggests that the PMTC program can be effective in adults with mental disabilities, especially in reducing gingival inflammation.
Assuntos
Assistência Odontológica para a Pessoa com Deficiência , Profilaxia Dentária , Deficiência Intelectual , Adulto , Bactérias/crescimento & desenvolvimento , Distribuição de Qui-Quadrado , Suscetibilidade à Cárie Dentária , Depósitos Dentários/prevenção & controle , Depósitos Dentários/terapia , Placa Dentária/microbiologia , Feminino , Seguimentos , Hemorragia Gengival/prevenção & controle , Hemorragia Gengival/terapia , Gengivite/prevenção & controle , Gengivite/terapia , Humanos , Concentração de Íons de Hidrogênio , Masculino , Índice de Higiene Oral , Bolsa Periodontal/prevenção & controle , Bolsa Periodontal/terapia , Periodontite/terapia , Estatísticas não ParamétricasRESUMO
The distribution of carbonic anhydrase isozyme II (CA II)-like immunoreactivity (-LI) in the gustatory epithelium was examined in the adult rat. In the circumvallate and foliate papillae, CA II-LI was observed in the cytoplasm of the spindle-shaped taste bud cells, with weak immunoreaction in the surface of the gustatory epithelium. No neuronal elements displayed CA II-LI in these papillae. There was no apparent difference in the distribution pattern between the anterior and posterior portions of the foliate papillae. In immunoelectron microscopy, immunoreaction products for CA II were diffusely distributed in the entire cytoplasm of the taste bud cells having dense round granules at the periphery of the cells. No taste bud cells displaying CA II-LI were detected in the fungiform papillae, but a few thick nerve fibers displayed CA II-LI. In the taste buds of the palatal epithelium, neither taste bud cells nor neuronal elements exhibited CA II-LI. The present results indicate that CA II was localized in the type I cells designated as supporting cells in the taste buds located in the posterior lingual papillae of the adult animal.
Assuntos
Anidrases Carbônicas/análise , Isoenzimas/análise , Palato/enzimologia , Língua/enzimologia , Animais , Anidrases Carbônicas/química , Epitélio/enzimologia , Imuno-Histoquímica , Isoenzimas/química , Masculino , Palato Mole/enzimologia , Ratos , Ratos Sprague-Dawley , Papilas Gustativas/enzimologiaRESUMO
Fimbriae of Porphyromonas gingivalis are thought to play an important role in the colonization and invasion of periodontal tissues. In this study, we analyzed the interactions of P. gingivalis fimbriae with human hemoglobin, fibrinogen, and salivary components (i.e., proline-rich protein [PRP], proline-rich glycoprotein [PRG], and statherin) based on surface plasmon resonance (SPR) spectroscopy with a biomolecular interaction analyzing system (BIAcore). The real-time observation showed that the fimbriae interacted more quickly with hemoglobin and PRG than with other proteins and more intensely with fibrinogen. The significant association constant (ka) values obtained by BIAcore demonstrated that the interactions between fimbriae and these host proteins are specific. These estimated Ka values were not too different; however, the Ka values for hemoglobin (2.43 x 10(6)) and fibrinogen (2.16 x 10(6)) were statistically greater than those for the salivary proteins (1.48 x 10(6) to 1.63 x 10(6)). The Ka value of anti-fimbriae immunoglobulin G for fimbriae was estimated to be 1. 22 x 10(7), which was 6.55-fold higher than the mean Ka value of the host proteins. Peptide PRP-C, a potent inhibitor of PRP-fimbriae interaction, dramatically inhibited fimbrial association to PRP and PRG and was also inhibitory against other host proteins by BIAcore. The binding of fimbriae to these proteins was also evaluated by other methods with hydroxyapatite beads or polystyrene microtiter plates. The estimated binding abilities differed considerably, depending on the assay method that was used. It was noted that the binding capacity of PRP was strongly diminished by immobilization on a polystyrene surface. Taken together, these findings suggest that P. gingivalis fimbriae possess a strong ability to interact with the host proteins which promote bacterial adherence to the oral cavity and that SPR spectroscopy is a useful method for analyzing specific protein-fimbriae interactions.
Assuntos
Fímbrias Bacterianas/fisiologia , Porphyromonas gingivalis/fisiologia , Porphyromonas gingivalis/patogenicidade , Proteínas/metabolismo , Sequência de Aminoácidos , Animais , Anticorpos Antibacterianos , Aderência Bacteriana/fisiologia , Infecções por Bacteroidaceae/etiologia , Fibrinogênio/metabolismo , Fímbrias Bacterianas/imunologia , Hemoglobinas/metabolismo , Humanos , Técnicas In Vitro , Cinética , Dados de Sequência Molecular , Peptídeos/química , Peptídeos/metabolismo , Doenças Periodontais/etiologia , Porphyromonas gingivalis/imunologia , Domínios Proteicos Ricos em Prolina , Ligação Proteica , Coelhos , Proteínas e Peptídeos Salivares/química , Proteínas e Peptídeos Salivares/metabolismo , Ressonância de Plasmônio de Superfície , Virulência/fisiologiaRESUMO
Fimbriae (FimA) of Porphyromonas gingivalis are filamentous components on the cell surface and are thought to play an important role in the colonization and invasion of periodontal tissues. We previously demonstrated that fimA can be classified into four variants (types I to IV) on the basis of the nucleotide sequences of the fimA gene. In the present study, we attempted to detect the four different fimA genes in saliva and plaque samples isolated from patients with periodontitis using the PCR method. Four sets of fimA type-specific primers were designed for the PCR assay. These primers selectively amplified 392-bp (type I), 257-bp (type II), 247-bp (type III), and 251-bp (type IV) DNA fragments of the fimA gene. Positive PCR results were observed with reference strains of P. gingivalis in a type-specific manner. All other laboratory strains of oral and nonoral bacteria gave negative results. The sensitivity of the PCR assay for fimA type-specific detection was between 5 and 50 cells of P. gingivalis. Clinical samples were obtained from saliva and subgingival plaque from deep pockets (>/=4 mm) of 93 patients with periodontitis. Bacterial genomic DNA was isolated from the samples, and the targeted fragments were amplified by PCR. The presence of P. gingivalis was demonstrated in 73 patients (78.5%), and a single fimA gene was detected in most patients. The distribution of the four fimA types among the P. gingivalis-positive patients was as follows: type I, 5.4%; type II, 58.9%; type III, 6. 8%; type IV, 12.3%; types I and II, 6.8%; types II and IV, 2.7%; and untypeable, 6.8%. P. gingivalis with type II fimA was detected more frequently in the deeper pockets, and a significant difference of the occurrence was observed between shallow (4 mm) and deep (>/=8 mm) pockets. These results suggest that P. gingivalis strains that possess type II fimA are significantly more predominant in periodontitis patients, and we speculate that these organisms are involved in the destructive progression of periodontal diseases.
Assuntos
Proteínas de Bactérias/genética , Proteínas de Fímbrias , Periodontite/microbiologia , Porphyromonas gingivalis/isolamento & purificação , Adulto , Idoso , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Porphyromonas gingivalis/genética , Sensibilidade e EspecificidadeRESUMO
This study was conducted to identify bilirubin in deciduous teeth obtained from two patients with a history of severe liver dysfunction. Teeth were histologically analyzed and bilirubin was extracted and quantified spectrophotometrically. Histological analysis revealed a green line in the dentine running parallel to the incremental lines. A chloroform/methanol/acetic acid (30:10:0.5, v/v) extract of the teeth was evaporated and the residue dissolved in chloroform. Absorption spectra were prepared before and after the diazo reaction. The absorption maximum shifted from 450 nm before to 540 nm after the diazo reaction and was higher than that of normal deciduous teeth. These results indicate that the discolouration of teeth in patients with severe liver dysfunction is due to bilirubin deposition.
Assuntos
Bilirrubina/análise , Hiperbilirrubinemia/complicações , Descoloração de Dente/etiologia , Estudos de Casos e Controles , Criança , Dente Canino/patologia , Humanos , Incisivo/patologia , Masculino , Dente Decíduo/patologiaRESUMO
OBJECTIVE: Hereditary sensory and autonomic neuropathy type IV (congenital insensitivity to pain with anhidrosis) is a rare disorder. In this study, we investigated the oral and dental manifestations associated with hereditary sensory and autonomic neuropathy type IV. STUDY DESIGN: Eighteen patients with hereditary sensory and autonomic neuropathy type IV whose ages ranged from 1 year 0 months to 22 years 3 months were examined for oral signs and symptoms of tooth abnormalities, malocclusions, soft tissue disorders, tongue papilla atrophy, and morphologic abnormalities of hands and fingers. RESULTS: All 18 patients showed congenital insensitivity to pain and anhidrosis. Oral self-mutilations, such as autoextraction of teeth and severe biting injuries (with resultant scarring) of the finger tips and oral soft tissues (tongue, lip, and buccal mucosa), were found in most patients. In infant patients the condition was typically characterized by decubital ulcers on the ventral surface of the tongue, resulting from trauma of the incisal edge of erupting mandibular primary incisors during sucking or nursing. These ulcers led to several local and systemic problems, such as tongue bleeding, infection, malnutrition, and halitosis. A large number of missing teeth and a high incidence of dental caries were additional characteristic findings. Such oral self-mutilations were found to decrease with age and with the intellectual, social, and/or emotional development of the patients. However, not all of the mutilations were completely eliminated. Two patients had partial dentures to replace missing teeth. CONCLUSIONS: Our study suggests that early diagnosis and specific dental management for patients with hereditary sensory and autonomic neuropathy type IV are important for prevention of the characteristic oral and dental problems accompanying this disorder.