RESUMEN
INTRODUCTION: Paget's disease of bone (PDB) is a skeletal disorder characterized by disorganized bone remodeling due to abnormal osteoclasts. Tumor necrosis factor receptor superfamily member 11A (TNFRSF11A) gene encodes the receptor activator of nuclear factor kappa B (RANK), which has a critical role in osteoclast function. There are five types of rare PDB and related osteolytic disorders due to TNFRSF11A tandem duplication variants so far, including familial expansile osteolysis (84dup18), expansile skeletal hyperphosphatasia (84dup15), early-onset familial PDB (77dup27), juvenile PDB (87dup15), and panostotic expansile bone disease (90dup12). MATERIALS AND METHODS: We reviewed a Japanese family with PDB, and performed whole-genome sequencing to identify a causative variant. RESULTS: This family had bone symptoms, hyperphosphatasia, hearing loss, tooth loss, and ocular manifestations such as angioid streaks or early-onset glaucoma. We identified a novel duplication variant of TNFRSF11A (72dup27). Angioid streaks were recognized in Juvenile Paget's disease due to loss-of-function variants in the gene TNFRSF11B, and thought to be specific for this disease. However, the novel recognition of angioid streaks in our family raised the possibility of occurrence even in bone disorders due to TNFRSF11A duplication variants and the association of RANKL-RANK signal pathway as the pathogenesis. Glaucoma has conversely not been reported in any case of Paget's disease. It is not certain whether glaucoma is coincidental or specific for PDB with 72dup27. CONCLUSION: Our new findings might suggest a broad spectrum of phenotypes in bone disorders with TNFRSF11A duplication variants.
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Estrías Angioides , Glaucoma , Osteítis Deformante , Humanos , Receptor Activador del Factor Nuclear kappa-B/genética , Osteítis Deformante/genéticaRESUMEN
OBJECTIVES: The aims of this study were (i) to compare the osteogenic impact of low-intensity pulsed ultrasound (LIPUS) and low-magnitude high-frequency (LMHF) loading achieved with whole-body vibration (WBV) on peri-implant bone healing and implant osseointegration in rat tibiae, and (ii) to examine their combined effect on these processes. MATERIAL AND METHODS: Titanium implants were inserted in the bilateral tibiae of 28 Wistar rats. Rats were randomly divided into four groups: LIPUS + WBV, LIPUS, WBV, and control. LIPUS was applied to the implant placement site for 20 min/day on 5 days/week (1.5 MHz and 30 mW/cm2 ). WBV was applied for 15 min/day on 5 days/week (50 Hz and 0.5 g). In the LIPUS + WBV group, both stimuli were applied under the same stimulation conditions as in the LIPUS and WBV groups. After 4 weeks of treatment, peri-implant bone healing and implant osseointegration were assessed using removal torque (RT) tests, micro-CT analyses of relative gray (RG) value, and histomorphometrical analyses of bone-to-implant contact (BIC) and peri-implant bone formation (BV/TV). RESULTS: The LIPUS + WBV group had significantly greater BIC than the WBV and control groups. Although there were no significant intergroup differences in RT, RG value, and BV/TV, these variables tended to be greater in the LIPUS + WBV group than the other groups. CONCLUSIONS: The combination of LIPUS and LMHF loading may promote osteogenic activity around the implant. However, further study of the stimulation conditions of LIPUS and LMHF loading is necessary to better understand the osteogenic effects and the relationship between the two stimuli.
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Implantes Dentales , Osteogénesis , Animales , Oseointegración , Ratas , Ratas Wistar , Ondas Ultrasónicas , VibraciónRESUMEN
PURPOSE: To examine the effect of tightening and loosening the dental implant screw on implant osseointegration in an elderly rat model. MATERIALS AND METHODS: Titanium dental implants were inserted in the bilateral tibiae of 34 elderly Wistar rats (1 year and 3 months old). Rats were randomly divided into five groups: control (no loading); immediate loading (IL) (vertical load [3 Hz for 15 minutes/day] immediately after implantation); early loading (EL) (load started on postoperative day 7); five times abutment removal (R5) (tightening and loosening of the abutment screw once per day for 5 days each week); and two times abutment screw removal (R2) (tightening and loosening of the abutment screw once per day for 2 days each week). After 4 weeks, the rats were euthanized, and the bone structure surrounding the dental implants was evaluated using microcomputed tomography analysis. RESULTS: Osseointegration failed more frequently in the EL, R2, and R5 groups than in the control and IL groups (P = .06). The EL, R2, and R5 groups also tended to have lower relative gray values than the control and IL groups. CONCLUSION: Tightening and loosening the abutment screw might negatively affect dental implant osseointegration in the early healing stage in older adults.
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Pilares Dentales , Implantes Dentales , Animales , Tornillos Óseos , Análisis del Estrés Dental , Oseointegración , Ratas , Ratas Wistar , Torque , Microtomografía por Rayos XRESUMEN
OBJECTIVE: It remains to be elucidated whether and how endothelial functions are impaired in peripheral circulation of patients with coronary functional disorders, such as vasospastic angina (VSA) and microvascular angina (MVA). We simultaneously examined endothelial functions of peripheral conduit and resistance arteries in patients with coronary functional disorders, with a special reference to NO and endothelium-dependent hyperpolarization factors. Approach and Results: Based on the results of invasive coronary acetylcholine testing and coronary physiological measurements, we divided 43 patients into 3 groups; VSA, MVA, and VSA+MVA. Endothelium-dependent vasodilatations of the brachial artery and fingertip arterioles to intra-arterial infusion of bradykinin were simultaneously evaluated by ultrasonography and peripheral arterial tonometry, respectively. To assess NO and endothelium-dependent hyperpolarization factors, measurements were repeated after oral aspirin and intra-arterial infusion of NG-monomethyl-L-arginine. Additionally, endothelium-independent vasodilatations to sublingual nitroglycerin and plasma levels of biomarkers for endothelial functions were measured. Surprisingly, digital vasodilatations to bradykinin were almost absent in patients with MVA alone and those with VSA+MVA compared with those with VSA alone. Mechanistically, both NO- and endothelium-dependent hyperpolarization-mediated digital vasodilatations were markedly impaired in patients with MVA alone. In contrast, endothelium-independent vasodilatations to nitroglycerin were comparable among the 3 groups. Plasma levels of soluble VCAM (vascular cell adhesion molecule)-1 were significantly higher in patients with MVA alone compared with those with VSA alone. CONCLUSIONS: These results provide the first evidence that both NO- and endothelium-dependent hyperpolarization-mediated digital vasodilatations are markedly impaired in MVA patients, suggesting that MVA is a cardiac manifestation of the systemic small artery disease.
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Arteriolas/fisiopatología , Arteria Braquial/fisiopatología , Endotelio Vascular/fisiopatología , Dedos/irrigación sanguínea , Angina Microvascular/fisiopatología , Enfermedad Arterial Periférica/fisiopatología , Vasodilatación , Anciano , Arteriolas/efectos de los fármacos , Arteriolas/metabolismo , Factores Biológicos/metabolismo , Arteria Braquial/efectos de los fármacos , Arteria Braquial/metabolismo , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/metabolismo , Femenino , Humanos , Masculino , Angina Microvascular/diagnóstico , Persona de Mediana Edad , Óxido Nítrico/metabolismo , Enfermedad Arterial Periférica/diagnóstico , Resistencia Vascular , Vasodilatación/efectos de los fármacos , Vasodilatadores/administración & dosificaciónRESUMEN
Endothelium-dependent hyperpolarization (EDH) factor is one of endothelium-derived relaxing factors and plays important roles especially in microvessels. We have previously demonstrated that endothelium-derived hydrogen peroxide (H2O2) is an EDH factor produced by all types of nitric oxide synthases (NOSs), including endothelial NOS (eNOS), neuronal NOS (nNOS), and inducible NOS. Recent studies have suggested the association between coronary microvascular dysfunction and cardiac diastolic dysfunction. However, the role of EDH in this issue remains to be fully elucidated. We thus examined whether EDH plays an important role in coronary microcirculation and if so, whether endothelial dysfunction, especially impaired EDH, is involved in the pathogenesis of cardiac diastolic dysfunction in mice. Using a Langendorff-perfused heart experiment, we examined the increase in coronary flow in response to bradykinin in the presence of indomethacin and N-nitro-L-arginine (EDH condition) in wild-type, eNOS-knockout (KO), and nNOS/eNOS-double-KO mice. Compared with wild-type mice, EDH-mediated relaxations were increased in eNOS-KO mice but were significantly reduced in n/eNOS-KO mice. Catalase, a specific H2O2 scavenger, markedly inhibited EDH-mediated relaxations in all 3 genotypes, indicating compensatory roles of nNOS-derived H2O2 as an EDH factor in coronary microcirculation. Although both eNOS-KO and n/eNOS-KO mice exhibited similar extents of cardiac morphological changes, only n/eNOS-KO mice exhibited cardiac diastolic dysfunction. The expression of oxidized protein kinase G I-α (PKGIα) in the heart was significantly increased in eNOS-KO mice compared with n/eNOS-KO mice. These results indicate that EDH/H2O2 plays important roles in maintaining coronary microcirculation and cardiac diastolic function through oxidative PKGIα activation.
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Factores Biológicos/metabolismo , Circulación Coronaria , Vasos Coronarios/metabolismo , Endotelio Vascular/metabolismo , Hipertrofia Ventricular Izquierda/metabolismo , Microcirculación , Microvasos/metabolismo , Vasodilatación , Disfunción Ventricular Izquierda/metabolismo , Función Ventricular Izquierda , Animales , Proteínas de Unión al Calcio/metabolismo , Vasos Coronarios/fisiopatología , Proteína Quinasa Dependiente de GMP Cíclico Tipo I/metabolismo , Diástole , Endotelio Vascular/fisiopatología , Peróxido de Hidrógeno/metabolismo , Hipertrofia Ventricular Izquierda/genética , Hipertrofia Ventricular Izquierda/fisiopatología , Preparación de Corazón Aislado , Masculino , Ratones Endogámicos C57BL , Ratones Noqueados , Microvasos/fisiopatología , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo I/genética , Óxido Nítrico Sintasa de Tipo I/metabolismo , Óxido Nítrico Sintasa de Tipo III/genética , Óxido Nítrico Sintasa de Tipo III/metabolismo , Oxidación-Reducción , Fosforilación , Transducción de Señal , Disfunción Ventricular Izquierda/genética , Disfunción Ventricular Izquierda/fisiopatologíaRESUMEN
Although increased levels of reactive oxygen species (ROS) are involved in the pathogenesis of cardiovascular diseases, the importance of physiological ROS has also been emerging. We have previously demonstrated that endothelium-derived H2O2 is an endothelium-dependent hyperpolarization (EDH) factor and that loss of endothelial caveolin-1 reduces EDH/H2O2 in the microcirculation. Caveolin-1 (Cav-1) is a scaffolding/regulatory protein that interacts with diverse signaling pathways, including angiogenesis. However, it remains unclear whether endothelial Cav-1 plays a role in ischemic angiogenesis by modulating EDH/H2O2. In the present study, we thus addressed this issue in a mouse model of hindlimb ischemia using male endothelium-specific Cav-1 (eCav-1) knockout (KO) mice. In isometric tension experiments with femoral arteries from eCav-1-KO mice, reduced EDH-mediated relaxations to acetylcholine and desensitization of sodium nitroprusside-mediated endothelium-independent relaxations were noted ( n = 4~6). An ex vivo aortic ring assay also showed that the extent of microvessel sprouting was significantly reduced in eCav-1-KO mice compared with wild-type (WT) littermates ( n = 12 each). Blood flow recovery at 4 wk assessed with a laser speckle flowmeter after femoral artery ligation was significantly impaired in eCav-1-KO mice compared with WT littermates ( n = 10 each) and was associated with reduced capillary density and muscle fibrosis in the legs ( n = 6 each). Importantly, posttranslational protein modifications by reactive nitrogen species and ROS, as evaluated by thiol glutathione adducts and nitrotyrosine, respectively, were both increased in eCav-1-KO mice ( n = 6~7 each). These results indicate that endothelial Cav-1 plays an important role in EDH-mediated vasodilatation and ischemic angiogenesis through posttranslational protein modifications by nitrooxidative stress in mice in vivo. NEW & NOTEWORTHY Although increased levels of reactive oxygen species (ROS) are involved in the pathogenesis of cardiovascular diseases, the importance of physiological ROS has also been emerging. The present study provides a line of novel evidence that endothelial caveolin-1 plays important roles in endothelium-dependent hyperpolarization and ischemic angiogenesis in hindlimb ischemia in mice through posttranslational protein modifications by reactive nitrogen species and ROS in mice in vivo.
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Caveolina 1/metabolismo , Endotelio Vascular/metabolismo , Neovascularización Patológica/fisiopatología , Animales , Caveolina 1/genética , Arteria Femoral/fisiopatología , Miembro Posterior/irrigación sanguínea , Peróxido de Hidrógeno/metabolismo , Isquemia/fisiopatología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Estrés Oxidativo , Procesamiento Proteico-Postraduccional , Especies de Nitrógeno Reactivo , Especies Reactivas de Oxígeno/metabolismo , Flujo Sanguíneo Regional , VasodilataciónRESUMEN
AIMS: Nitric oxide (NO) and endothelium-dependent hyperpolarization (EDH) play important roles in maintaining cardiovascular homeostasis. We have previously demonstrated that endothelial NO synthase (eNOS) plays diverse roles depending on vessel size, as a NO generating system in conduit arteries and an EDH-mediated system in resistance arteries, for which caveolin-1 (Cav-1) is involved. However, the physiological role of endothelial Cav-1 in microvessels remains to be elucidated. METHODS AND RESULTS: We newly generated endothelium-specific Cav-1-knockout (eCav-1-KO) mice. eCav-1-KO mice showed loss of endothelial Cav-1/eNOS complex and had cardiac hypertrophy despite normal blood pressure. In eCav-1-KO mice, as compared to wild-type controls, the extent of eNOS phosphorylation at inhibitory Thr495 was significantly reduced in mesenteric arteries and the heart. Isometric tension and Langendorff-perfused heart experiments showed that NO-mediated responses were enhanced, whereas EDH-mediated responses were reduced in coronary microcirculation in eCav-1-KO mice. Immunohistochemistry showed increased level of 8-nitroguanosine 3',5'-cyclic monophosphate (8-nitro-cGMP), a marker of nitrative stress, in the heart from eCav-1-KO mice. S-guanylation of cardiac H-Ras in eCav-1-KO mice was also significantly increased compared with wild-type controls. CONCLUSIONS: These results suggest that eCav-1 is involved in the protective role of EDH against nitrative stress caused by excessive NO to maintain cardiac microvascular homeostasis.
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Factores Biológicos/farmacología , Cardiomegalia/metabolismo , Caveolina 1/metabolismo , Vasos Coronarios/efectos de los fármacos , Células Endoteliales/efectos de los fármacos , Arterias Mesentéricas/efectos de los fármacos , Microvasos/efectos de los fármacos , Donantes de Óxido Nítrico/farmacología , Óxido Nítrico/metabolismo , Estrés Nitrosativo , Vasodilatadores/farmacología , Animales , Factores Biológicos/metabolismo , Cardiomegalia/genética , Cardiomegalia/patología , Cardiomegalia/fisiopatología , Caveolina 1/deficiencia , Caveolina 1/genética , Vasos Coronarios/metabolismo , Vasos Coronarios/fisiopatología , GMP Cíclico/análogos & derivados , GMP Cíclico/metabolismo , Células Endoteliales/metabolismo , Guanosina/análogos & derivados , Guanosina/metabolismo , Preparación de Corazón Aislado , Masculino , Arterias Mesentéricas/metabolismo , Arterias Mesentéricas/fisiopatología , Ratones Endogámicos C57BL , Ratones Noqueados , Microvasos/metabolismo , Microvasos/fisiopatología , Donantes de Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo III/metabolismo , Nitrocompuestos/metabolismo , Fosforilación , Procesamiento Proteico-Postraduccional , Proteínas Proto-Oncogénicas p21(ras)/metabolismo , Transducción de Señal/efectos de los fármacosRESUMEN
Endothelium-dependent hyperpolarization (EDH) plays important roles in the systemic circulation, whereas its role in the pulmonary circulation remains largely unknown. Furthermore, the underlying mechanisms of pulmonary hypertension (PH) also remain to be elucidated. We thus aimed to elucidate the role of EDH in pulmonary circulation in general and in PH in particular. In isolated perfused lung and using male wild-type mice, endothelium-dependent relaxation to bradykinin (BK) was significantly reduced in the presence of Nω-nitro-l-arginine by ~50% compared with those in the presence of indomethacin, and the combination of apamin plus charybdotoxin abolished the residual relaxation, showing the comparable contributions of nitric oxide (NO) and EDH in the pulmonary microcirculation under physiological conditions. Catalase markedly inhibited EDH-mediated relaxation, indicating the predominant contribution of endothelium-derived H2O2. BK-mediated relaxation was significantly reduced at day 1 of hypoxia, whereas it thereafter remained unchanged until day 28. EDH-mediated relaxation was diminished at day 2 of hypoxia, indicating a transition from EDH to NO in BK-mediated relaxation before the development of hypoxia-induced PH. Mechanistically, chronic hypoxia enhanced endothelial NO synthase expression and activity associated with downregulation of caveolin-1. Nitrotyrosine levels were significantly higher in vascular smooth muscle of pulmonary microvessels under chronic hypoxia than under normoxia. A similar transition of the mediators in BK-mediated relaxation was also noted in the Sugen hypoxia mouse model. These results indicate that EDH plays important roles in the pulmonary microcirculation in addition to NO under normoxic conditions and that impaired EDH-mediated relaxation and subsequent nitrosative stress may be potential triggers of the onset of PH. NEW & NOTEWORTHY This study provides novel evidence that both endothelium-dependent hyperpolarization and nitric oxide play important roles in endothelium-dependent relaxation in the pulmonary microcirculation under physiological conditions in mice and that hypoxia first impairs endothelium-dependent hyperpolarization-mediated relaxation, with compensatory upregulation of nitric oxide, before the development of hypoxia-induced pulmonary hypertension.
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Factores Biológicos/metabolismo , Endotelio Vascular/fisiopatología , Hipertensión Pulmonar/etiología , Hipoxia/complicaciones , Microcirculación , Arteria Pulmonar/fisiopatología , Circulación Pulmonar , Vasodilatación , Animales , Modelos Animales de Enfermedad , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/metabolismo , Hipertensión Pulmonar/metabolismo , Hipertensión Pulmonar/fisiopatología , Hipoxia/metabolismo , Hipoxia/fisiopatología , Masculino , Potenciales de la Membrana , Ratones Endogámicos C57BL , Microcirculación/efectos de los fármacos , Óxido Nítrico/metabolismo , Arteria Pulmonar/efectos de los fármacos , Circulación Pulmonar/efectos de los fármacos , Transducción de Señal , Tirosina/análogos & derivados , Tirosina/metabolismo , Remodelación Vascular , Vasodilatación/efectos de los fármacos , Vasodilatadores/farmacologíaRESUMEN
Nickel, cobalt, and chromium are well known to be causal agents of allergic contact dermatitis. Palladium (Pd) can also cause allergic disease and exposure results from wide use of this metal in dental restorations and jewelry. Metal allergy is categorized as a delayed-type hypersensitivity, and metal-responsive T cell clones have been isolated from allergic patients. However, compared to nickel, little is known about the pathology of allergic disease mediated by Pd, and pathogenic T cells are poorly understood. To identify the pathogenic T cells that are responsible for onset of Pd allergy, we enriched metal-responsive lymphocytes by sequential adoptive transfer of involved lymph node cells. Here we show that sequential adoptive transfer gradually increased the incidence and the intensity of Pd allergy, and CD8⺠T cells are responsible for the disease as CD8⺠T cell-depleted mice and ß2-microglobulin-deficient mice did not develop Pd allergy. In addition, we found that draining lymph node cells skewed toward CD8⺠T cells in response to Pd challenge in 8th adoptive transferred recipient mice. The CD8⺠T cells expressed NKG2D, a costimulatory molecule involved in the production of IFN-γ. NKG2D ligand was also induced in Pd-injected tissues. Furthermore, both NKG2D ligand-transgenic mice, where NKG2D is downmodulated, and IFN-γ-deficient mice showed impaired Pd allergy. Taken together, these results indicate that IFN-γ-producing NKG2D⺠CD8⺠T cells are responsible for Pd allergy and suggest that NKG2D is a potential therapeutic target for treatment of metal allergy.
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Linfocitos T CD8-positivos/inmunología , Dermatitis por Contacto/inmunología , Interferón gamma/metabolismo , Subfamilia K de Receptores Similares a Lectina de Células NK/metabolismo , Paladio/inmunología , Traslado Adoptivo , Animales , Ligandos , Metales/inmunología , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Desnudos , Ratones Transgénicos , Microglobulina beta-2/metabolismoRESUMEN
This study aimed to investigate the prevalence and levels of major periodontal pathogens, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythia in subgingival plaque samples of a group of Japanese patients with aggressive periodontitis (AgP) and chronic periodontitis (CP). A total of 40 patients with clinical diagnosis of AgP or CP and 10 periodontally healthy volunteers were subjected to clinical and microbiological analysis. Subgingival plaque samples were analyzed for A. actinomycetemcomitans, P. gingivalis and T. forsythia with a real-time polymerase chain reaction (PCR) technique. The prevalence of P. gingivalis and T. forsythia was relatively high in patients with periodontitis: over 60% of AgP or CP patients harbored these pathogens whereas they were not detected in the subgingival plaque samples from periodontally healthy individuals. P. gingivalis and T. forsythia were relatively frequently detected together in AgP and CP patients. No significant differences in the prevalence or level of the 3 pathogens were found between periodontitis groups. The proportion of T. forsythia was approximately 4-fold higher in CP group than in AgP group (P = 0.02). In periodontitis patients, a significant positive correlation was found between periodontal parameters (probing depth and clinical attachment level) and the numbers of total bacteria, P. gingivalis and T. forsythia. No distinct pattern of the subgingival profile of these pathogens was discerned between the two disease entities, except for the difference in the proportion of T. forsythia. The red complex bacteria, P. gingivalis and T. forsythia were highly prevalent in this population of Japanese AgP and CP patients, collaborating their roles in periodontitis.
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Aggregatibacter actinomycetemcomitans/aislamiento & purificación , Periodontitis Agresiva , Bacteroidaceae/aislamiento & purificación , Periodontitis Crónica , Porphyromonas gingivalis/aislamiento & purificación , Adulto , Periodontitis Agresiva/epidemiología , Periodontitis Agresiva/microbiología , Infecciones Bacterianas/epidemiología , Infecciones Bacterianas/microbiología , Periodontitis Crónica/epidemiología , Periodontitis Crónica/microbiología , Femenino , Humanos , Japón/epidemiología , Masculino , Persona de Mediana Edad , PrevalenciaRESUMEN
We report a case involving a 12-year follow-up after treatment for chronic periodontitis with furcation involvement. A 54-year-old woman presented with the chief complaint of hypersensitivity. Clinical examination at the first visit revealed 15% of sites with a probing depth ≥4 mm and 35% of sites with bleeding on probing. Initial periodontal therapy was implemented based on a clinical diagnosis of severe chronic periodontitis. Surgical periodontal therapy was subsequently performed at selected sites. For #44, regenerative periodontal therapy using enamel matrix derivative (Emdogain(®)) was selected. For #16, which exhibited a 2- to 3-wall vertical bony defect and class III (mesio-distal) furcation involvement, bone graft was scheduled. Other sites with residual periodontal pockets were treated by open flap debridement. For #37, with a gutter-shaped root, odontoplasty was performed. After reevaluation, the patient was placed on supportive periodontal therapy (SPT). During 12 years of SPT, the periodontal condition remained uneventful in most of the teeth. However, bone resorption was observed in the distal aspect of #37, making the prognosis poor. This indicates the need to continuously monitor risk factors, including inflammation and traumatic occlusion, during SPT. Although some problems still remain, severe periodontitis with furcation involvement was successfully maintained longitudinally with an adequate level of patient compliance and careful SPT.
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Periodontitis Crónica/prevención & control , Defectos de Furcación/prevención & control , Pérdida de Hueso Alveolar/prevención & control , Trasplante Óseo/métodos , Desbridamiento/métodos , Proteínas del Esmalte Dental/uso terapéutico , Oclusión Dental Traumática/prevención & control , Placa Dental/prevención & control , Femenino , Estudios de Seguimiento , Hemorragia Gingival/prevención & control , Recesión Gingival/prevención & control , Regeneración Tisular Guiada Periodontal/métodos , Humanos , Estudios Longitudinales , Persona de Mediana Edad , Desbridamiento Periodontal/métodos , Bolsa Periodontal/prevención & control , Factores de Riesgo , Colgajos Quirúrgicos/cirugíaRESUMEN
Periodontopathic bacteria such as Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Tannerella forsythia, Campylobacter rectus and Treponema denticola play an important role in the initiation and progression of periodontitis. The aim of this investigation was to evaluate the relationship between periodontal clinical parameters and the subgingival profile of periodontopathic bacteria. Twenty-six periodontitis patients (23-62 years of age; mean age, 40.2±13.2) with no systemic disease agreed to participate in the study. Periodontal clinical parameters, including probing depth (PD) and bleeding on probing (BOP) were recorded. Subgingival plaque samples were obtained from deep (PD≥4 mm) and shallow (PD≤3 mm) pockets in each patient for detection of P. gingivalis, A. actinomycetemcomitans, T. forsythia, C. rectus and T. denticola by polymerase chain reaction technique. The relationship between the periodontal pathogens and clinical parameters was determined with the Fisher exact test, and a statistically significant association was found between detection of P. gingivalis, T. forsythia, C. rectus and T. denticola and PD or BOP. T. denticola was the most prevalent pathogen in both shallow PD and deep PD sites. No statistically significant association was found between detection of A. actinomycetemcomitans and the clinical parameters examined. A statistically significant association was found between detection of the red complex bacteria and the clinical parameters. These results suggest that the red complex pathogens and C. rectus play an important role in the initiation and progression of periodontitis.
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Periodontitis Crónica/microbiología , Placa Dental/microbiología , Bolsa Periodontal/microbiología , Adulto , Aggregatibacter actinomycetemcomitans/genética , Aggregatibacter actinomycetemcomitans/aislamiento & purificación , Técnicas de Tipificación Bacteriana , Bacteroides/genética , Bacteroides/aislamiento & purificación , Campylobacter rectus/genética , Campylobacter rectus/aislamiento & purificación , Distribución de Chi-Cuadrado , ADN Bacteriano/análisis , Femenino , Humanos , Masculino , Persona de Mediana Edad , Índice Periodontal , Porphyromonas gingivalis/genética , Porphyromonas gingivalis/aislamiento & purificación , Treponema denticola/genética , Treponema denticola/aislamiento & purificación , Adulto JovenRESUMEN
BACKGROUND: Antimicrobial proteins are abundant in saliva. The purpose of this study was to determine and compare the amounts of two types of antibacterial protein, cystatin and lysozyme, in saliva between healthy persons and subjects with periodontitis. METHODS: Forty subjects with periodontitis visiting Tokyo Dental College Chiba Hospital, Chiba, Japan, and 27 healthy persons were evaluated. Whole saliva was collected by requiring all subjects to expectorate into a sterile tube. Salivary levels of cystatin SA, cystatin C, and lysozyme were determined by enzyme-linked immunosorbent assay or immunoblot assay. RESULTS: Cystatin SA levels in saliva from the periodontally diseased group showed a mean value of 0.063 +/- 0.026 mg/ml, statistically lower than that in the healthy group (P <0.05). The average cystatin C level in the periodontally diseased group was 2.27 +/- 1.20 ng/ml, markedly lower than that in the healthy group (3.79 +/- 1.28 ng/ml; P <0.05). Average lysozyme levels in the periodontitis and healthy groups were 16.75 +/- 15.31 microg/ml and 30.03 +/- 15.03 microg/ml, respectively. The lysozyme level in the periodontitis group was significantly lower than in the healthy group (P <0.05). CONCLUSION: Specific monoclonal antibodies are useful for the detection of family 2 cystatins in saliva samples, and the amount of antibacterial protein in saliva offers a potential indicator of the risk for periodontitis.
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Cistatinas/análisis , Muramidasa/análisis , Periodontitis/enzimología , Proteínas y Péptidos Salivales/análisis , Adulto , Anciano , Anciano de 80 o más Años , Estudios de Casos y Controles , Cistatina C , Cisteína Endopeptidasas/metabolismo , Femenino , Humanos , Immunoblotting , Masculino , Persona de Mediana Edad , Inhibidores de Proteasas/análisis , Cistatinas SalivalesRESUMEN
To develop a diagnostic trial enabling the selective examination for a target cystatin in human body fluids, we attempted to prepare monoclonal antibodies against human cystatin SA1 (originally cystatin SA) and its variant form (cystatin SA2). BALB/c mice were immunized with recombinant (r-) cystatins SA1 and SA2. Two monoclonal antibodies designated Cys3F11 and Cys2E5 were selected. By ELISA analyses, the Cys2E5 was shown to react with r-cystatin SA2 but also somewhat with r-cystatin SA1 (22% cross-reactivity) and with plasma cystatin C (18% cross-reactivity), indicating a high specificity for cystatin SA2. The Cys3F11 reacted not only with r-cystatin SA1 but also with r-cystatin SA2 (89% cross-reactivity) and plasma cystatin C (47% cross-reactivity). This finding was further emphasized by immunoblotting of human submandibular-sublingual saliva samples. ELISA additivity test suggests that the two monoclonal antibodies bind to distinct epitopes. In conclusion, we have succeeded in producing two antibodies that discriminate the structural differences between salivary cystatins S and SN, which share more than 90% identity in amino acid sequence with cystatin SA.
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Anticuerpos Monoclonales/inmunología , Cistatinas/química , Cistatinas/genética , Cistatinas/inmunología , Secuencia de Aminoácidos , Animales , Western Blotting , Reacciones Cruzadas , Cistatinas/aislamiento & purificación , Ensayo de Inmunoadsorción Enzimática , Epítopos , Femenino , Variación Genética , Hibridomas/inmunología , Ratones , Ratones Endogámicos BALB C , Datos de Secuencia Molecular , Proteínas Recombinantes/química , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/aislamiento & purificación , Saliva/química , Cistatinas Salivales , Homología de Secuencia de AminoácidoRESUMEN
Periodontitis is a common inflammatory disease causing destruction of periodontal tissues. It is a multifactor disease involving genetic factors and oral environmental factors. To determine genetic risk factors associated with aggressive periodontitis or severe chronic periodontitis, single nucleotide polymorphisms (SNPs) in multiple candidate genes were investigated in Japanese. We studied 134 patients with aggressive periodontitis, 117 patients with severe chronic periodontitis, and 125 healthy volunteers without periodontitis, under case-control setting, and 310 SNPs in 125 candidate genes were genotyped. Association evaluation by Fisher's exact test (p < 0.01) revealed statistically significant SNPs in multiple genes, not only in inflammatory mediators (IL6ST and PTGDS, associated with aggressive periodontitis; and CTSD, associated with severe chronic periodontitis), but also in structural factors of periodontal tissues (COL4A1, COL1A1, and KRT23, associated with aggressive periodontitis; and HSPG2, COL17A1, and EGF, associated with severe chronic periodontitis). These appear to be good candidates as genetic factors for future study.