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1.
Curr Issues Mol Biol ; 45(2): 1306-1313, 2023 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-36826030

RESUMO

Oral mucositis is highly prevalent among the elderly, for whom oral care is often difficult. Oral mucositis, such as candidiasis, can induce systemic fungemia. Antifungal prophylaxis may be useful in such cases to prevent systemic fungemia; however, studies on this are limited. The objective of this study was to demonstrate the effectiveness of antifungal prophylaxis to prevent systemic Candida dissemination compared to oral care using a mice model. Oral candidiasis was induced using chemotherapy and inoculation with C. albicans in 8-week-old male mice. Group A was given oral care, Group B was orally administered an antifungal drug, Group C was intravenously administered an antifungal drug, and Group D was used as the negative control group. Macroscopic features of the tongue surface, colony forming units (CFU) on the tongue, and blood culture for C. albicans were evaluated. CFU was significantly higher in Group A than in Groups B and C. The oral care group, but not the groups administered antifungal agents, showed significantly higher positive numbers of animals with C. albicans in the blood as compared to the control group, indicating the effectiveness of antifungal prophylaxis over oral care. Antifungal prophylaxis may be an option for the prevention of systemic fungemia in individuals with difficulty in oral care.

2.
J Oral Implantol ; 49(2): 187-196, 2023 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-36796077

RESUMO

The bone ring technique is applied for vertical augmentation, in which the dental implant and autogenous block bone graft are simultaneously inserted. We investigated the healing of bone around implants placed simultaneously using the bone ring technique with and without membrane placement after a 12-month healing period. Vertical bone defects were created on both sides of the mandible of Beagle dogs. Implants were inserted into the defects through bone rings and fixed with membrane screws as healing caps. The augmented sites on one side of the mandible were covered with a collagen membrane. Samples were harvested 12 months after implantation and assessed histologically and by microcomputed tomography analysis. All implants remained throughout the healing period; however, except for 1 implant, they showed lost caps and/or exposure to the oral cavity. The implants had contact with newly formed bone despite frequent bone resorption. The surrounding bone appeared mature. The medians of bone volume and percentages of total bone area and bone-to-implant contact within the bone ring were slightly higher in the group with membrane placement than in that without membrane placement. Nevertheless, none of the evaluated parameters were significantly affected by the membrane placement. In the present model, soft tissue complication was frequent, and the membrane application did not reveal the effect at 12 months after implant placement using the bone ring technique. Sustained osseointegration and maturation of surrounding bone were observed in both groups after a 12-month healing period.


Assuntos
Implantação Dentária Endóssea , Implantes Dentários , Cães , Animais , Implantação Dentária Endóssea/métodos , Microtomografia por Raio-X , Osseointegração , Mandíbula/transplante , Transplante Ósseo/métodos
3.
Ann Plast Surg ; 89(2): 218-224, 2022 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-35276708

RESUMO

PURPOSE: Gradual elevation of periosteum from the bone surface is known to promote the adaptation of soft tissues and the formation of hard tissues. The aim of our study was to estimate the benefit of periosteal distraction osteogenesis (PDO) on de novo bone formation in a rat model. MATERIALS AND METHODS: After device placement, animals were allowed for a latency period of 7 days. Animals in the PDO group were subjected to distraction at a rate of 0.1 mm/d for 10 days. In the periosteal pumping (PP) group, the animals were subjected to distraction at a rate of 0.1 mm/d. The direction of distraction was alternated every 2 days. The animals were euthanized at 17, 31, and 45 days after surgery, and the samples were analyzed histologically and by microcomputed tomography. RESULTS: In both groups, the new bone was characterized as primary woven bone that was located at the leading edge of bone apposition. Bone volumes significantly increased throughout the observation period both in the PP group ( P = 0.018) and in the PDO group ( P < 0.001). The new bone was denser and more mature in the PP group than in the PDO group, and the difference was significant at the 31-day time point ( P = 0.024). However, the volume of the new bone was higher in the PDO at the 45-day time point ( P < 0.001). CONCLUSIONS: We propose that the PP may be applied to enhance the osteogenic capacity of periosteum without plate elevation. Because this is only a proof-of-principle study, the alternated protocol of periosteal distraction warrants evaluation in the future studies.


Assuntos
Osteogênese por Distração , Periósteo , Animais , Estudos de Viabilidade , Osteogênese , Osteogênese por Distração/métodos , Periósteo/cirurgia , Ratos , Crânio/cirurgia , Microtomografia por Raio-X
4.
Odontology ; 109(1): 82-102, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32681298

RESUMO

Currently, it is difficult to extract exosomes with stable physicochemical properties from saliva. Furthermore, due to inadequate availability of basic data, the application of salivary exosomes as a diagnostic material is limited. In this study, we aimed to investigate an easier method for extraction of exosomes from whole saliva and compared proteins in salivary exosomes derived from subjects of two age groups. Salivary exosomes were extracted from nine females (56.7 ± 1.17 years old; climacteric or 19.9 ± 0.20 years old; adolescent) using commercial reagents and kits and detected using western blotting with anti-exosome marker antibodies. Exosome particle size and exosome-containing proteins were identified using NanoSight® and liquid chromatography with tandem mass spectrometry, respectively. In addition, an efficient method of exosome extraction from saliva using a reagent and without the use of an ultracentrifuge was shown. Our results showed a higher total protein content and larger particle size in climacteric exosomes than in adolescent exosomes. However, adolescent exosomes showed a larger variety of proteins (780 proteins) than the climacteric exosomes (573 proteins). Altogether, 893 proteins were identified in the salivary exosomes. Although viral process-, ribosome- and structural molecule-related proteins were higher in the adolescent exosomes, the levels of major salivary proteins such as immunoglobulins and amylase, were higher in the climacteric exosomes than in the adolescent exosomes. The data presented, which show the fundamental protein composition of salivary exosomes and the changes that occur with age, are beneficial in both diagnostic and biotechnological applications.


Assuntos
Climatério , Exossomos , Adolescente , Adulto , Feminino , Humanos , Pessoa de Meia-Idade , Proteínas , Saliva , Espectrometria de Massas em Tandem , Adulto Jovem
5.
Clin Oral Implants Res ; 31(1): 29-36, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31541500

RESUMO

OBJECTIVES: To assess the impact of collagen membrane application on bone formation surrounding implants placed simultaneously with the bone ring technique. MATERIAL AND METHODS: Dental implants were inserted simultaneously with the bone ring technique in standardized, vertical alveolar bony defects in the mandible of dogs. On one side of the mandible, the augmented sites were covered with a collagen membrane (M-Group). On the contralateral side, no membranes were used (NM-Group). Implants were left to heal with osseointegration for three and six months. The harvested samples were analyzed by means of micro-CT. RESULTS: A nonparametric analysis of data revealed that the membranes were not a significant negative factor for bone volume (BV), but for bone-to-implant contact (BIC, p = .04). Absence of healing caps impaired BV (p = .04) and BIC (p = .02) as well. Furthermore, loss of healing caps and exposure to the oral environment significantly and negatively affected BV (p < .001) and bone mineral density (p < .05) within 2 mm below the implant shoulder. Implant exposure and healing time had a negative interaction effect on both BV (p = .01) and BIC (p = .01). CONCLUSIONS: Within its limitations, the present study revealed no benefit of membrane application to implant placement simultaneous with the bone ring technique. Disruption of soft tissue healing was identified as a risk factor for decrease in BV and BIC.


Assuntos
Implantação Dentária Endóssea , Implantes Dentários , Animais , Cães , Mandíbula , Osseointegração , Microtomografia por Raio-X
6.
Clin Oral Implants Res ; 29(3): 263-276, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29235158

RESUMO

AIM: To evaluate the impact of a collagen membrane on bone remodeling and osseointegration of implants placed simultaneously with a bone ring technique. MATERIAL AND METHODS: Standardized, vertical alveolar bone defects in the mandibles of six dogs were created. Tapered dental implants designed for two-stage subcrestal placement were inserted simultaneously with a bone ring technique. On one side of the mandible, the augmented sites were covered with a collagenous membrane. Implants with (M Group) and without membranes (NM Group) were left for an osseointegration period of 3 and 6 months, respectively. Block biopsies of the implants with surrounding bone were harvested and analyzed histologically. RESULTS: Implant exposure was a common finding (2/3) concomitantly with loss of healing caps. It appeared to be related to advanced bone loss around the implants. Exposure of implants was more frequent in M Group, however, without significant differences when compared to NM Group. The total bone area within the region of the bone ring was greater in the NM Group compared to the M Group. Moreover, in the region of the pristine bone of the M Group, the total bone was greater than at the corresponding NM Group sites at both observation periods. A nonparametric analysis of variance (ANOVA) revealed no significant effects of membrane placement or healing period on the total area of the bone. The total bone-to-implant contact (BIC) for the two groups was similar at each observation time point. However, BIC increased significantly at 6-month compared with 3-month observation period (p = .0088) in both groups. CONCLUSIONS: In vertical bone augmentation applying the bone ring technique, the disruption of soft tissue was a frequent complication. Membrane placement yielded no significant advantage on the osseointegration (BIC) of implants or bone characteristics.


Assuntos
Remodelação Óssea , Transplante Ósseo/métodos , Implantação Dentária Endóssea/métodos , Implantes Dentários , Mandíbula/cirurgia , Osseointegração , Perda do Osso Alveolar/patologia , Aumento do Rebordo Alveolar/métodos , Análise de Variância , Animais , Cães , Regeneração Tecidual Guiada , Masculino , Mandíbula/diagnóstico por imagem , Mandíbula/patologia , Membranas Artificiais , Modelos Animais , Estatísticas não Paramétricas
7.
J Craniofac Surg ; 29(2): 376-381, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29084111

RESUMO

Antiseptic solutions are commonly utilized to treat local infection in the oral and maxillofacial region. However, surrounding vital bone is also exposed to antiseptic agents during irrigation and may have a potential negative impact on bone survival. The aim of the present study was therefore to investigate the effect of rinsing time with various antiseptic solutions on bone cell viability, as well as their subsequent release of growth factors important for bone regeneration. The bone samples collected from porcine mandible were rinsed in the following commonly utilized antiseptic solutions; povidone-iodine (0.5%), chlorhexidine digluconate (CHX, 0.2%), hydrogen peroxide (1%), and sodium hypochlorite (0.25%) for 1, 5, 10, 20, 30, or 60 minutes and assessed for cell viability and release of growth factors including vascular endothelial growth factor, transforming growth factor beta 1, bone morphogenetic protein 2, receptor activator of nuclear factor kappa-B ligand, and interleukin-1 beta by enzyme-linked immunosorbent assay. It was found in all the tested groups that the long exposure of any of the tested antiseptic solutions drastically promoted higher cell death. Sodium hypochlorite demonstrated the significantly highest cell death and at all time points. Interestingly, bone cell viability was highest in the CHX group post short-term rinsing of 1, 5, or 10 minutes when compared with the other 4 tested groups. A similar trend was also observed in subsequent growth factor release. The present study demonstrated that of the 4 tested antiseptic solutions, short-term CHX rinsing (ideally within 1 minute) favored bone cell viability and growth factor release. Clinical protocols should be adapted accordingly.


Assuntos
Anti-Infecciosos Locais/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Mandíbula/citologia , Animais , Células Cultivadas , Suínos , Fatores de Tempo
8.
Odontology ; 106(4): 398-407, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29557992

RESUMO

This study aimed to evaluate the impact of three types of block bone substitute material on bone formation and graft resorption in vivo. Standardized bone defects (n = 4 defects/animal) were created in the calvaria of nine dogs. Block bone substitutes made of deproteinized bovine bone mineral (DBBM), beta-tricalcium phosphate (ß-TCP) and a mixture alpha-TCP and hydroxyapatite (α-TCP/HA) were inserted into the bone defects. A fourth defect was left untreated (empty). All sites were covered with a collagenous membrane. Block biopsies were harvested at 3, 6 and 12 months post-implantation and analyzed by micro-CT and histology. Biomaterial absorption was minimal and incorporation within the defect margin was good for all biomaterials. However, ß-TCP demonstrated a relatively greater volume of new bone formation and less residual material volume when compared with DBBM and α-TCP/HA. Conversely, α-TCP/HA showed higher osteoconductive potential and a greater new bone area compared with the other two biomaterials. The block bone substitutes used in the present in vivo study showed advantageous in terms of maintenance of their original form in bony defect. However, the positive impact of all biomaterials on new bone formation and replacement of bone was minor even at 12 months. These findings indicate that block bone substitutes are not well suited to vertical bone augmentation. Further investigations are required to improve the insufficient new bone volume for promising clinical results.


Assuntos
Regeneração Óssea/efeitos dos fármacos , Substitutos Ósseos/farmacologia , Crânio/cirurgia , Animais , Materiais Biocompatíveis/farmacologia , Fosfatos de Cálcio/farmacologia , Bovinos , Cães , Durapatita/farmacologia , Masculino , Teste de Materiais , Minerais/farmacologia , Modelos Animais , Microtomografia por Raio-X
9.
Odontology ; 106(2): 117-124, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28748269

RESUMO

We previously showed that mRNA expression of BPIFB1 (Bpifb1), an antibacterial protein in the palate, lung, and nasal epithelium clone protein family, was increased in parotid acinar cells in non-obese diabetic (NOD, NOD/ShiJcl) mice, which is an animal model for Sjögren's syndrome. However, we did not previously assess the protein levels. In this report, we confirmed the expression of BPIFB1 protein in the parotid glands of NOD mice. Immunoblotting of subcellular fractions revealed that BPIBB1 was localised in secretory granules in parotid glands from NOD mice, and was almost not in parotid glands from the control mice. BPIFB1 had N-linked glycan that reacted with Aleuria aurantia lectin, which caused two types of spots with a slightly different pI and molecular weight. The expression of BPIFB1 protein was also demonstrated by immunohistochemistry. BPIFB1 was detected in the saliva from NOD mice but not in the saliva from the control mice, indicating individual constitution. BPIFB1 in saliva may be applied to other research as a diagnostic marker.


Assuntos
Proteínas de Transporte/metabolismo , Camundongos Endogâmicos NOD , Saliva/metabolismo , Animais , Diabetes Mellitus Experimental , Modelos Animais de Doenças , Immunoblotting , Imuno-Histoquímica , Camundongos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Eletroforese em Gel Diferencial Bidimensional
10.
Clin Oral Implants Res ; 28(7): e31-e38, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27090198

RESUMO

OBJECTIVES: The aim of this study was to evaluate the osseointegration of implants placed in a single-staged compared to two-staged procedure using bone ring technique. MATERIAL AND METHODS: In this study were used standardized, vertical alveolar bone defects in dogs. In the test group, dental implants (Straumann BL® , Basel, Switzerland) were inserted simultaneously with bone ring technique. As control group served implants inserted 6 months following grafting. Implants of both groups were left for an osseointegration period of 3 and 6 months. The peri-implant bone loss and bone-to-implant contact within the bone ring and native bone were analyzed morphometrically. An explorative statistical analysis was performed. RESULTS: The peri-implant bone level remained relatively stable within groups and between groups per given time period. Most of bone apposite on the implant surface in two groups was composed of newly formed bone. A nonparametric analysis of variance (ANOVA) revealed no significant advantage for two-staged implant placement for new and total bone, except for residual bone (P = .0084). Furthermore, two groups of implants performed similarly in bone ring and in native bone throughout the observation period. CONCLUSIONS: In terms of osseointegration, both techniques are likely equally efficient in the present defect model. The single-staged implant placement with cortical bone grafts warrants further documentation in clinical studies.


Assuntos
Aumento do Rebordo Alveolar/métodos , Transplante Ósseo/métodos , Implantação Dentária Endóssea/métodos , Implantes Dentários , Osseointegração/fisiologia , Perda do Osso Alveolar/patologia , Perda do Osso Alveolar/cirurgia , Animais , Cães , Masculino , Mandíbula/transplante , Osteotomia , Crânio/transplante , Retalhos Cirúrgicos , Microtomografia por Raio-X
11.
Odontology ; 105(3): 300-310, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27778135

RESUMO

We examined the effects of adhesive systems under study applied for a laser-cut cavity using an Er,Cr:YSGG laser on rat dental pulp at 24 h and 14 days postoperatively. Group 1, laser-cut cavities were treated with a self-etching-primer and bonding agent; group 2, pretreated with a phosphoric-acid, and then treated with a self-etching-primer and bonding agent; group 3, pretreated with a phosphoric-acid and sodium-hypochlorite, and then treated with a self-etching-primer and bonding agent; and group 4, treated with an all-in-one adhesive. A flowable resin composite was used as filling material for each cavity treated with each group. A glass-ionomer-cement was used as a control. The following items were evaluated: pulp-tissue-disorganization (PTD), inflammatory-cell-infiltration (ICI), tertiary-dentin-formation (TDF), and bacterial-penetration (BP). The results were statistically analyzed using the Kruskal-Wallis test and Mann-Whitney U test. No significant differences were observed among the experimental groups for all parameters after 24 h and 14 days (P > 0.05). The majority of the specimens showed PTD with edema formation after 24 h; however, all the specimens demonstrated pulpal healing with TDF after 14 days. On the parameter of TDF, all groups showed significant differences between the two postoperative periods (P < 0.01). On the parameter of ICI, a significant difference was found between the two postoperative periods in group 4 (P < 0.05). No specimens showed BP. The pretreatment on the cavity prepared with the laser using phosphoric-acid or sodium-hypochlorite did not affect the dental pulp healing of rat tooth.


Assuntos
Colagem Dentária/métodos , Preparo da Cavidade Dentária/métodos , Polpa Dentária/efeitos da radiação , Adesivos Dentinários/química , Lasers de Estado Sólido/uso terapêutico , Condicionamento Ácido do Dente , Animais , Cromo , Resinas Compostas/química , Érbio , Gálio , Cimentos de Ionômeros de Vidro/química , Masculino , Ratos , Ratos Sprague-Dawley , Cimentos de Resina , Escândio , Ítrio
12.
Clin Oral Implants Res ; 27(11): 1384-1391, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26833804

RESUMO

BACKGROUND: Simultaneous implant placement with bone grafting shortens the overall treatment period, but might lead to the peri-implant bone loss or even implant failure. The aim of this study was to compare the single-staged to two-staged implant placement using the bone ring technique. MATERIAL AND METHODS: Four standardized alveolar bone defects were made in the mandibles of nine dogs. Dental implants (Straumann BL® , Basel, Switzerland) were inserted simultaneously with bone ring technique in test group and after 6 months of healing period in control group. Animals of both groups were euthanized at 3 and 6 months of osseointegration period. The harvested samples were analyzed by means of histology and micro-CT. RESULTS: The amount of residual bone decreased while the amount of new bone increased up to 9 months of healing period. All morphometric parameters remained stable between 3 and 6 months of osseointegration period within groups. Per a given time point, median area of residual bone graft was higher in test group and area of new bone in control group. The volume of bone ring was greater in test than in control group, reaching the significance at 6 months of osseointegration period (P = 0.002). CONCLUSIONS: In the present type of bone defect, single-staged implant placement may be potentially useful to shorten an overall treatment period.


Assuntos
Aumento do Rebordo Alveolar/métodos , Transplante Ósseo/métodos , Implantação Dentária Endóssea/métodos , Implantes Dentários , Osseointegração , Perda do Osso Alveolar/patologia , Perda do Osso Alveolar/cirurgia , Animais , Cães , Masculino , Mandíbula/transplante , Osteotomia , Crânio/transplante , Retalhos Cirúrgicos , Microtomografia por Raio-X
13.
Odontology ; 102(1): 98-104, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23263522

RESUMO

Mouse molars undergo distal movement, during which new bone is formed at the mesial side of the tooth root whereas the preexisting bone is resorbed at the distal side of the root. However, there is little detailed information available regarding which of the bones that surround the tooth root are involved in physiological tooth movement. In the present study, we therefore aimed to investigate the precise morphological differences of the alveolar bone between the bone formation side of the tooth root, using routine histological procedures including silver impregnation, as well as by immunohistochemical analysis of alkaline phosphatase and tartrate-resistant acid phosphatase activity, and immunohistochemical analysis of the expression of the osteocyte markers dentin matrix protein 1, sclerostin, and fibroblast growth factor 23. Histochemical analysis indicated that bone formation by osteoblasts and bone resorption by osteoclasts occurred at the bone formation side and the bone resorption side, respectively. Osteocyte marker immunoreactivity of osteocytes at the surface of the bone close to the periodontal ligament differed at the bone formation and bone resorption sides. We also showed different specific features of osteocytic lacunar canalicular systems at the bone formation and bone resorption sides by using silver staining. This study suggests that the alveolar bone is different in the osteocyte nature between the bone formation side and the bone resorption side due to physiological distal movement of the mouse molar.


Assuntos
Processo Alveolar/fisiologia , Dente Molar/fisiologia , Técnicas de Movimentação Dentária , Animais , Imuno-Histoquímica , Masculino , Camundongos
14.
Histochem Cell Biol ; 140(5): 567-74, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23604549

RESUMO

An immunohistochemical examination of guinea-pig taste buds in vallate papillae revealed gustducin-immunoreactive cells in the area of von Ebner's glands, minor salivary glands. Since there have been no reports describing those cells in these locations for other species, we investigated these glands in order both to localize the cells and compare their immunoreactive characteristics with corresponding cells in the vallate taste buds. The gustducin-immunoreactive cells coincided with cells containing no secretory granules in the end portion of the glands, which was supported by the electron-microscopic immunocytochemistry. Double immunofluorescence microscopy confirmed these cells to be entirely immunopositive to type III inositol 1,4,5-triphosphate receptor (IP3R-3), phospholipase Cß2 (PLCß2), and villin and also partly immunopositive to neuron-specific enolase (NSE) and calbindin D-28K. The gustducin-immunoreactive cells in the vallate taste buds exhibited completely the same immunoreactivities for these five molecules. Accordingly, the present results give credence to a consideration that the gustducin-immunnoreactive cells in both locations are identical in function(s) e.g., chemo-reception.


Assuntos
Papilas Gustativas/citologia , Transducina/análise , Transducina/imunologia , Glândulas de von Ebner/citologia , Animais , Feminino , Cobaias , Imuno-Histoquímica , Masculino , Papilas Gustativas/imunologia , Glândulas de von Ebner/imunologia
15.
Arch Biochem Biophys ; 530(1): 32-9, 2013 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-23262319

RESUMO

Cystatin D encoded by Cst5 is a salivary classified type II cystatin. We investigated the dynamism of cystatin D by examining the distribution of cystatin D protein and mRNA in rats, to identify novel functions. The simultaneous expression of Cst5 and cystatin D was observed in parotid glands, however in situ hybridization showed that only acinar cells produced cystatin D. Synthesized cystatin D was localized in small vesicles and secreted from the apical side to the saliva, and from the basolateral side to the extracellular region, a second secretory pathway for cystatin D. We also identified antigen-presenting cells in the parotid glands that contained cystatin D without the expression of Cst5, indicating the uptake of cystatin D from the extracellular region. Cystatin D was detected in blood serum and renal tubular cells with megalin, indicating the circulation of cystatin D through the body and uptake by renal tubular cells. Thus, the novel dynamism of cystatin D was shown and a function for cystatin D in the regulation of antigen-presenting cell activity was proposed.


Assuntos
Células Acinares/metabolismo , Células Apresentadoras de Antígenos/metabolismo , Cistatinas/metabolismo , Glândula Parótida/citologia , Sequência de Aminoácidos , Animais , Cistatinas/química , Cistatinas/genética , Regulação da Expressão Gênica , Túbulos Renais/citologia , Masculino , Transporte Proteico , Ratos , Ratos Wistar
16.
J Biol Chem ; 286(39): 33854-62, 2011 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-21832089

RESUMO

Rab27, a small GTPase, is generally recognized as an important regulator of secretion that interacts with Rab27-specific effectors to regulate events in a wide variety of cells, including endocrine and exocrine cells. However, the mechanisms governing the spatio-temporal regulation of GTPase activity of Rab27 are not firmly established, and no GTPase-activating protein (GAP) specific for Rab27 has been identified in secretory cells. We previously showed that expression of EPI64, a Tre-2/Bub2/Cdc16 (TBC)-domain-containing protein, in melanocytes inactivates endogenous Rab27A on melanosomes (Itoh, T., and Fukuda, M. (2006) J. Biol. Chem. 281, 31823-31831), but the EPI64 role in secretory cells has never been investigated. In this study, we investigated the effect of EPI64 on Rab27 in isoproterenol (IPR)-stimulated amylase release from rat parotid acinar cells. Subcellular fractionation and immunohistochemical analyses indicated that EPI64 was enriched on the apical plasma membrane of parotid acinar cells. We found that an antibody against the TBC/Rab-GAP domain of EPI64 inhibited the reduction in levels of the endogenous GTP-Rab27 in streptolysin-O-permeabilized parotid acinar cells and suppressed amylase release in a dose-dependent manner. We also found that the levels of EPI64 mRNA and EPI64 protein increased after IPR stimulation, and that treatment with actinomycin D or antisense-EPI64 oligonucleotides suppressed the increase of EPI64 mRNA/EPI64 protein and the amount of amylase released. Our findings indicated that EPI64 acted as a physiological Rab27-GAP that enhanced GTPase activity of Rab27 in response to IPR stimulation, and that this activity is required for IPR-induced amylase release.


Assuntos
Amilases/metabolismo , Membrana Celular/metabolismo , Glândula Parótida/metabolismo , Proteínas/metabolismo , Proteínas rab de Ligação ao GTP/metabolismo , Animais , Células Cultivadas , Dactinomicina/farmacologia , Relação Dose-Resposta a Droga , Isoproterenol/farmacologia , Inibidores da Síntese de Ácido Nucleico/farmacologia , Glândula Parótida/citologia , RNA Mensageiro/metabolismo , Ratos , Simpatomiméticos/farmacologia
17.
Eur J Oral Sci ; 120(2): 123-31, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22409218

RESUMO

Exocytosis of secretory granules in parotid acinar cells requires multiple events: tethering, docking, priming, and fusion with a luminal plasma membrane. The exocyst complex, which is composed of eight subunits (Sec3, Sec5, Sec6, Sec8, Sec10, Sec15, Exo70, and Exo84) that are conserved in yeast and mammalian cells, is thought to participate in the exocytotic pathway. However, to date, no exocyst subunit has been identified in salivary glands. In the present study, we investigated the expression and function of exocyst subunits in rat parotid acinar cells. The expression of mRNA for all eight exocyst subunits was detected in parotid acinar cells by RT-PCR, and Sec6 and Sec8 proteins were localized on the luminal plasma membrane. Sec6 interacted with Sec8 after 5 min of stimulation with isoproterenol. In addition, antibodies to-Sec6 and Sec8 inhibited isoproterenol-induced amylase release from streptolysin O-permeabilized parotid acinar cells. These results suggest that an exocyst complex of eight subunits is required for amylase release from parotid acinar cells.


Assuntos
Células Acinares/enzimologia , Amilases/metabolismo , Isoproterenol/farmacologia , Proteínas de Membrana Transportadoras/fisiologia , Glândula Parótida/enzimologia , Simpatomiméticos/farmacologia , Células Acinares/efeitos dos fármacos , Animais , Fracionamento Celular , Exocitose/fisiologia , Proteínas de Membrana Transportadoras/genética , Glândula Parótida/citologia , Glândula Parótida/efeitos dos fármacos , Ratos
18.
Imaging Sci Dent ; 51(1): 73-80, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33828964

RESUMO

PURPOSE: The purpose of this study was to evaluate the cone-beam computed tomographic (CBCT) imaging and histopathological characteristics of osteoradionecrosis (ORN) and medication-related osteonecrosis of the jaw (MRONJ). MATERIALS AND METHODS: Ten surgical specimens from segmental mandibulectomy (3 ORN and 7 MRONJ) were analyzed using CBCT. The CBCT parameters were as follows: high-resolution mode (tube voltage, 90.0 kV; tube current, 4.00 mA; rotation time, 16.8 s; field of view, 56 mm×56 mm; thickness, 0.099 mm). Histopathological characteristics were evaluated using histological slides of the surgical specimens. The Pearson chi-square test was used to compare ORN and MRONJ in terms of CBCT findings (internal texture, sequestrum, periosteal reaction and cortical perforation) and histopathological characteristics (necrotic bone, inflammatory cells, reactive bone formation, bacteria, Actinomyces, and osteoclasts). A P value less than 0.05 was considered to indicate statistical significance. RESULTS: MRONJ showed periosteal reaction on CBCT more frequently than ORN (7 of 7 [100%] vs. 0 of 3 [0%], P<0.05). Regarding histopathological characteristics, MRONJ showed osteoclasts more frequently than ORN (6 of 7 [85.7%] vs. 0 of 3 [0%], P<0.05). CONCLUSION: This study evaluated the CBCT imaging and histopathological characteristics of ORN and MRONJ, and the findings suggest that CBCT could be useful for the evaluation of ORN and MRONJ.

19.
Imaging Sci Dent ; 49(4): 281-286, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31915613

RESUMO

PURPOSE: The purpose of this study was to evaluate the morphology of the mandibular cortex in cases of medication-related osteonecrosis of the jaw (MRONJ) in patients with osteoporosis or bone metastases using a computer programme. MATERIALS AND METHODS: Fifty-four patients with MRONJ (35 with osteoporosis and 19 with bone metastases) were examined using panoramic radiography. The morphology of the mandibular cortex was evaluated using a computer programme that scanned the mandibular inferior cortex and automatically assessed the mandibular cortical index (MCI) according to the thickness and roughness of the mandibular cortex, as follows: normal (class 1), mildly to moderately eroded (class 2), or severely eroded (class 3). The MCI classifications of MRONJ patients with osteoporosis or bone metastases were evaluated with the Pearson chi-square test. In these analyses, a 5% significance level was used. RESULTS: The MCI of MRONJ patients with osteoporosis (class 1: 6, class 2: 15, class 3: 14) tended to be higher than that of patients with bone metastases (class 1: 14, class 2: 5, class 3: 0) (P=0.000). CONCLUSION: The use of a computer programme to assess mandibular cortex morphology may be an effective technique for the objective and quantitative evaluation of the MCI in MRONJ patients with osteoporosis or bone metastases.

20.
Ann Nucl Med ; 33(10): 776-782, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31342316

RESUMO

OBJECTIVE: To investigate the quantitative SPECT/CT imaging for medication-related osteonecrosis of the jaw (MRONJ) using volume-based parameters. METHODS: Thirteen patients with mandibular lesions (nine MRONJ and four chronic osteomyelitis) underwent SPECT/CT acquisition at 4 h after injection of technetium 99m hydroxymethylene diphosphonate. Then, reconstruction of the acquired data and underwent voxel-based quantitative analysis using GI-BONE software. The parameters of the quantitative manner, such as maximum standardized uptake value (SUVmax), peak SUV (SUVpeak), mean SUV (SUVmean), metabolic bone volume (MBV) and total bone uptake (TBU), were compared for the mandibular lesions. Statistical analyses for the quantitative parameters of mandibular lesions were performed by Mann-Whitney U test. A p value lower than 0.05 was considered as statistically significant. RESULTS: Then, reconstruction of these quantitative parameters, SUVmax (10.16 ± 0.96), SUVpeak (7.95 ± 0.88), SUVmean (5.90 ± 0.86) and TBU (94.22 ± 57.44) for chronic osteomyelitis were significantly higher than those for MRONJ [5.50 ± 2.47 (p = 0.020), 4.10 ± 1.85 (p = 0.011), 2.74 ± 1.07 (p = 0.006) and 29.88 ± 15.46 (p = 0.034), respectively]. CONCLUSIONS: SUVmax, SUVpeak, SUVmean and TBU derived from bone SPECT/CT and voxel-based quantitative parameters may be useful for the evaluation of mandibular lesions, such as MRONJ and chronic osteomyelitis.


Assuntos
Osteonecrose da Arcada Osseodentária Associada a Difosfonatos/diagnóstico por imagem , Osteomielite/diagnóstico por imagem , Tomografia Computadorizada com Tomografia Computadorizada de Emissão de Fóton Único , Adulto , Idoso , Idoso de 80 Anos ou mais , Doença Crônica , Feminino , Humanos , Processamento de Imagem Assistida por Computador , Masculino , Pessoa de Meia-Idade
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