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1.
Arch Oral Biol ; 97: 42-51, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30342306

RESUMO

OBJECTIVES: Gingival recession and alveolar bone loss are common manifestations of periodontitis. Periodontal regeneration is the ideal strategy for rehabilitating periodontal tissue defects and preventing tooth loss. The present study examined whether localized, topical application of gingival overgrowth-inducing drugs, phenytoin, nifedipine or cyclosporine, induces periodontal regeneration. METHODS: Polylactic-co-glycolic acid (PLGA) was used as the carrier for preparation of phenytoin, nifedipine or cyclosporine-loaded PLGA microspheres, using an oil-in-water emulsification technique. The drug-loaded microspheres were delivered to periodontal defects created on alveolar ridges mesial to the first maxillary molars of Sprague-Dawley rats. After eight weeks, the operation area in each rat, including the maxillary molars and periodontal tissues, was harvested and evaluated by micro-computed tomography, histochemical and immunohistochemical analyses. RESULTS: Physical parameters representative of periodontal regeneration, including the length of new alveolar bone (p < 0.01) and the area of new alveolar bone (p < 0.01) were significantly improved in the phenytoin group. Compared to other groups, the phenytoin group demonstrated increased expression of COL-1, VEGF-A, osteoblast and osteoclast markers (BMP-2, TGF-ß1, OCN and TRAP staining), as well as decreased expression of MMP-8. CONCLUSIONS: Results of the present study provided new evidence that localized, controlled release of phenytoin confers therapeutic benefits toward gingival recession and alveolar bone loss. Phenytoin appears to be a promising drug that promotes periodontal regeneration.


Assuntos
Perda do Osso Alveolar/tratamento farmacológico , Portadores de Fármacos/administração & dosagem , Retração Gengival/tratamento farmacológico , Microesferas , Nifedipino/administração & dosagem , Fenitoína/administração & dosagem , Poliésteres/administração & dosagem , Administração Tópica , Animais , Biomarcadores/análise , Ciclosporina/administração & dosagem , Imuno-Histoquímica , Masculino , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Método Simples-Cego , Microtomografia por Raio-X
2.
Inflamm Res ; 2018 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-30008029

RESUMO

AIM: The study aimed to investigate the effects of DNA repair proteins on cell apoptosis in human DPSCs during inflammation. METHODS: Lipopolysaccharide (LPS) was used to stimulate inflammation in dental pulp in vivo and in vitro. We identified the activation of DSB response and DNA repair proteins in inflamed pulp tissue and in LPS-treated human DPSCs. Then we transfected the cells with Ku70 (a key protein involved in NHEJ) siRNA and detected the expression changes of γ-H2A.X, DNA repair proteins and cell apoptosis. RESULTS: Immunohistochemical staining showed that at 4 and 6 days of pulpitis the expression of Ku70 and γ-H2A.X significantly increased. The levels of γ-H2A.X, Ku70, Xrcc4, and Rad51 increased considerably in the LPS-treated DPSCs. Furthermore, decreased expression of Ku70 could increase the number of γ-H2A.X foci, apoptotic cells and reduce cell viability in DPSCs. CONCLUSIONS: The results indicate that NHEJ pathway was the main mechanism involved in DNA damage response induced by repeated LPS stimulation in DPSCs. Meanwhile, the findings suggested that Ku70 serves importantly in the apoptosis of DPSCs in the inflammatory environment.

3.
J Endod ; 44(3): 414-431, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29273495

RESUMO

INTRODUCTION: The aim of this review was to evaluate whether the apical diameter of teeth with necrotic pulp affects the outcomes of regenerative endodontic treatment and to determine the minimal apical size needed to obtain proper pulp revascularization. METHODS: A literature search was performed from January 1, 2001, to November 25, 2016. Studies that satisfied the inclusion criteria were subjected to data extraction and analysis. RESULTS: In total, 14 studies with 85 patients were included. There were 10 case reports, 3 case series, and 1 prospective cohort study. The apical diameters of the teeth were divided into 3 groups: a narrow-sized group (group N), <0.5 mm (n = 10); a medium-sized group (group M), 0.5-1.0 mm (n = 25); and a wide-sized group (group W), >1.0 mm (n = 60). In group N, 1 tooth failed, 2 teeth completely healed, and 7 teeth incompletely healed. In group M, 2 teeth were excluded, and 1 tooth failed. In group W, 3 teeth were excluded, and 4 teeth failed. The clinical success rates were 90%, 95.65%, and 92.98% in groups N, M, and W, respectively. CONCLUSIONS: Within the limitations, the teeth with apical diameters <1.0 mm achieved clinical success after regenerative endodontic treatment. Meanwhile, the teeth with apical diameters of 0.5-1.0 mm attained the highest clinical success rate, which may be related to other potential factors, including patient age, pulp necrosis etiology, preoperative apical radiolucency, procedure details, follow-up period, and sample size.

4.
Environ Toxicol Pharmacol ; 51: 45-50, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28282589

RESUMO

This study investigated the correlation between differentially expressed proteins in amniotic fluid (AF) and cleft palate induced by all-trans retinoic acid (atRA), and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in mice. Seven proteins were differentially expressed at embryonic day (E) 16.5 in atRA and control groups as revealed by label-based mouse antibody array. Enzyme-linked immunosorbent assay was further used to detect the expression levels of these proteins in AF from E13.5 to E16.5 in atRA, TCDD, and control groups. The cleft palate groups showed lower concentrations of receptor for advanced glycation end products (RAGE) and epiregulin at E16.5. RAGE immunostaining obviously decreased in palatal tissue sections obtained from E14.5 to E16.5 in the cleft palate groups as revealed by immunohistochemistry. These findings indicate that reduced levels of RAGE and epiregulin in AF are correlated to chemically induced cleft palate in mice.


Assuntos
Líquido Amniótico/metabolismo , Fissura Palatina/metabolismo , Epirregulina/metabolismo , Receptor para Produtos Finais de Glicação Avançada/metabolismo , Animais , Biomarcadores/metabolismo , Fissura Palatina/induzido quimicamente , Modelos Animais de Doenças , Feminino , Idade Gestacional , Camundongos Endogâmicos C57BL , Palato/efeitos dos fármacos , Palato/embriologia , Palato/metabolismo , Dibenzodioxinas Policloradas/toxicidade , Gravidez , Receptor para Produtos Finais de Glicação Avançada/genética , Tretinoína/toxicidade
5.
Nat Commun ; 8: 14364, 2017 02 24.
Artigo em Inglês | MEDLINE | ID: mdl-28232668

RESUMO

Non-syndromic cleft lip with palate (NSCLP) is the most serious sub-phenotype of non-syndromic orofacial clefts (NSOFC), which are the most common craniofacial birth defects in humans. Here we conduct a GWAS of NSCLP with multiple independent replications, totalling 7,404 NSOFC cases and 16,059 controls from several ethnicities, to identify new NSCLP risk loci, and explore the genetic heterogeneity between sub-phenotypes of NSOFC. We identify 41 SNPs within 26 loci that achieve genome-wide significance, 14 of which are novel (RAD54B, TMEM19, KRT18, WNT9B, GSC/DICER1, PTCH1, RPS26, OFCC1/TFAP2A, TAF1B, FGF10, MSX1, LINC00640, FGFR1 and SPRY1). These 26 loci collectively account for 10.94% of the heritability for NSCLP in Chinese population. We find evidence of genetic heterogeneity between the sub-phenotypes of NSOFC and among different populations. This study substantially increases the number of genetic susceptibility loci for NSCLP and provides important insights into the genetic aetiology of this common craniofacial malformation.


Assuntos
Fenda Labial/genética , Fissura Palatina/genética , Loci Gênicos/genética , Predisposição Genética para Doença , Adulto , Fatores Etários , Grupo com Ancestrais do Continente Asiático/etnologia , Grupo com Ancestrais do Continente Asiático/genética , Estudos de Casos e Controles , Fenda Labial/etnologia , Fissura Palatina/etnologia , Estudos de Coortes , Feminino , Estudo de Associação Genômica Ampla , Técnicas de Genotipagem , Humanos , Masculino , Polimorfismo de Nucleotídeo Único , Fatores Sexuais
6.
Artigo em Inglês | MEDLINE | ID: mdl-28153567

RESUMO

OBJECTIVE: This study aims to analyze the expression of T-cell receptor γ chain alternate reading frame protein (TARP) in salivary adenoid cystic carcinoma (SACC) and its distant metastases and to investigate its influences on the development and progression of SACC. STUDY DESIGN: TARP expression was analyzed in 50 primary SACCs, 13 specimens of metastatic adenoid cystic carcinoma of salivary gland origin, and 20 noncancerous tissues around SACC via immunohistochemistry. Cell Counting Kit-8 tests, wound healing assay, and Transwell experiments were performed to evaluate the effects of lentivirus-mediated TARP overexpression on the proliferation, migration, and invasion of SACC cells. RESULTS: TARP expression was significantly increased in primary SACCs compared with adjacent noncancerous tissues, and this increase was further enhanced in metastases compared with primary SACCs. The expression level of TARP correlated significantly with tumor size, tumor-node-metastasis stage, perineural invasion, histologic type, and distant metastasis. Furthermore, TARP overexpression promoted the growth, migration, and invasion of SACC cells. CONCLUSIONS: TARP plays an important role in and may be used as a marker to indicate the development and progression of SACC.


Assuntos
Carcinoma Adenoide Cístico/metabolismo , Carcinoma Adenoide Cístico/patologia , Proteínas Nucleares/metabolismo , Neoplasias das Glândulas Salivares/metabolismo , Neoplasias das Glândulas Salivares/patologia , Adulto , Idoso , Western Blotting , Movimento Celular , Proliferação de Células , Eletroforese em Gel de Ágar , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica , Reação em Cadeia da Polimerase em Tempo Real
7.
Arch Oral Biol ; 60(3): 501-7, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25555252

RESUMO

OBJECTIVE: Glycogen synthase kinase-3ß (Gsk-3ß)/ß-catenin signaling regulates development of the secondary palate. It has been unclear about the effects of Gsk-3ß/ß-catenin signaling on palatal fusion and osteogenic differentiation in palatal shelves. DESIGN: In this study, palatal shelves from mouse embryonic day 13 (E13) were cultured in vitro with or without lithium chloride (LiCl). Palatal fusion was evaluated by haematoxylin-eosin staining. The expression of osteogenic markers in palatal shelves was measured by quantitative PCR, and immunohistochemical staining. Cell proliferation and apoptosis were examined by Ki-67 immunohistochemical and TUNEL staining, respectively. Gsk-3ß expression was evaluated by quantitative PCR and Western blotting. ß-catenin protein expression was evaluated by Western blotting. RESULTS: After the treatment with 10 mM LiCl, palatal shelves failed to fuse, and the mRNA and protein levels of osteogenic markers were reduced compared with controls. The number of Ki67-positive cell in the palatal osteoid was significantly higher in the LiCl group than in the controls. The apoptotic cells in the midline epithelial seam were reduced by LiCl. Gsk-3ß mRNA and protein expression levels decreased and ß-catenin protein expression levels increased by treatment of LiCl. CONCLUSION: Our findings show that LiCl-mediated GSK3ß inhibition prevents palatal fusion and osteogenic differentiation in palatal shelves by increased ß-catenin signaling. It indicated that overactivation of canonical Wnt signaling might impair the fusion of the secondary palate.


Assuntos
Fissura Palatina/induzido quimicamente , Cloreto de Lítio/toxicidade , Osteogênese/efeitos dos fármacos , Palato/efeitos dos fármacos , Palato/embriologia , Animais , Apoptose , Western Blotting , Proliferação de Células/efeitos dos fármacos , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Camundongos , Reação em Cadeia da Polimerase , RNA Mensageiro/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Proteínas Wnt/efeitos dos fármacos , beta Catenina/metabolismo
8.
J Cell Physiol ; 229(3): 384-92, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24037946

RESUMO

In the course of embryonic development skeletal elements form either through intramembranous or endochondral ossification. Wnt proteins play diverse roles during vertebrate skeletal development. Wnt16 is a key factor in developing long bones, but its exact role in craniofacial bone formation remains unclear. This study was initially undertaken to investigate the expression of Wnt16 during craniofacial bone development in mouse embryos. Wnt16 expression in the osteoid of calvaria, maxilla, and mandible started later than that of ALP and osteocalcin (OCN), but before mineralization of the craniofacial bones, suggesting that Wnt16 is involved in intramembranous ossification in the head. To confirm this, MC3T3-E1 cells were transfected with an adenovirus containing Wnt16 (Ad-Wnt16). Ad-Wnt16 cells showed decreased ALP activity and less mineralized nodule formations compared with control cells. In addition, the mRNA levels of osteogenic markers were reduced. Moreover, Wnt16 activated ß-catenin signaling in MC3T3-E1 cells at both transcription and protein levels as shown by a TOPflash luciferase reporter gene assay and western blot analysis. On the other hand, Wnt/ß-catenin pathway blockade by Dickkopf 1 abrogated the suppression of mineralization by Wnt16. Our findings suggest that Wnt16 is involved in intramembranous ossification and suppresses osteoblast differentiation through the Wnt/ß-catenin pathway.


Assuntos
Diferenciação Celular , Osteoblastos/metabolismo , Osteogênese , Crânio/metabolismo , Proteínas Wnt/metabolismo , Via de Sinalização Wnt , beta Catenina/metabolismo , Adenoviridae/genética , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Animais , Linhagem Celular , Regulação da Expressão Gênica no Desenvolvimento , Vetores Genéticos , Idade Gestacional , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Mandíbula/embriologia , Mandíbula/metabolismo , Maxila/embriologia , Maxila/metabolismo , Camundongos , Osteocalcina/genética , Osteocalcina/metabolismo , Crânio/embriologia , Fatores de Transcrição TCF/genética , Fatores de Transcrição TCF/metabolismo , Fatores de Tempo , Transfecção , Proteínas Wnt/genética , beta Catenina/genética
9.
Eur J Oral Sci ; 120(4): 278-82, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22813217

RESUMO

Tooth agenesis is one of the most common developmental disorders in humans. Previous studies have attributed non-syndromic tooth agenesis to mutations in several genes, including MSX1, PAX9, EDA, and AXIN2. In this study, we investigated a Chinese family with tooth agenesis combined with cleft lip. Genomic DNA was isolated from blood samples of all available family members. Candidate genes MSX1 and PAX9 were amplified by the PCR and directly sequenced. A novel heterozygous mutation at c.C565T, exon 2 of MSX1, was identified in affected members. To analyze the effect of the nonsense mutation on MSX1 expression, vectors containing wild-type and mutated MSX1 were constructed and transfected into COS7 cell lines. Real-time PCR showed that the mRNA expression of the mutated MSX1 was dramatically reduced compared with that of the wild-type MSX1. Our findings suggest that the nonsense mutation in MSX1 might have resulted in rapid degradation of the mutated transcript and caused the phenotype of tooth agenesis with cleft lip in the Chinese family.


Assuntos
Anodontia/genética , Fenda Labial/genética , Códon sem Sentido , Fator de Transcrição MSX1/genética , Fator de Transcrição PAX9/genética , Grupo com Ancestrais do Continente Asiático/genética , Estudos de Casos e Controles , China , Análise Mutacional de DNA , Feminino , Humanos , Fator de Transcrição MSX1/sangue , Fator de Transcrição MSX1/metabolismo , Masculino , Fator de Transcrição PAX9/sangue , Fator de Transcrição PAX9/metabolismo , Linhagem , Fenótipo , Reação em Cadeia da Polimerase em Tempo Real/métodos
10.
J Biomed Mater Res B Appl Biomater ; 100(5): 1435-43, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22287502

RESUMO

Periodontal regeneration is defined as regeneration of the tooth-supporting tissues including cementum, periodontal ligament, and alveolar bone. Guided tissue regeneration (GTR) has been demonstrated to be an effective technique to achieve periodontal regeneration. In the GTR procedures, various kinds of membranes play important roles. Chitosan, a deacetylated derivative of chitin, is biocompatible, biodegradable, and antimicrobial. It acts as hydrating agent and possesses tissue healing and osteoinducing effect. Chitosan can be easily processed into membranes, gels, nanofibers, beads, nanoparticles, scaffolds, and sponges forms and can be used in drug delivery systems. Here, we review the bioproperties of chitosan and report the progress of application of chitosan as membranes in GTR and guided bone regeneration (GBR), which indicates that chitosan could be a good substrate candidate as the materials for the GTR/GBR membranes.


Assuntos
Bioprótese , Quitosana , Regeneração Tecidual Guiada Periodontal/métodos , Membranas Artificiais , Periodonto , Regeneração , Animais , Regeneração Tecidual Guiada Periodontal/instrumentação , Humanos
11.
J Clin Periodontol ; 36(8): 627-33, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19552635

RESUMO

AIM: To clinically characterize and map the disease-associated locus in a five-generation Chinese family with autosomal dominant early-onset hereditary gingival fibromatosis (HGF). MATERIAL AND METHODS: A complete oral examination was conducted. Genomic DNA samples were obtained from 14 individuals. Short tandem repeats markers, which encompass four previously known loci related to HGF, were genotyped. Two-point log of the odds (LOD) scores were calculated using MLINK program of the LINKAGE software, multipoint and non-parametric linkage (NPL) analysis were performed using the GENEHUNTER software. RESULTS: Clinical evaluation and histological examination of this family suggested typical features of HGF. The onset age was early in the generations, ranging between 1 and 2 years. None of the tested markers showed cosegregation among affected individuals. Genotyping data from four putative regions yielded significant negative two-point LOD scores (<-2.0) at theta=0. The maximum multipoint LOD scores and NPL analysis revealed exclusion of these loci as well. CONCLUSIONS: Exclusion of linkage in this family to any of the known HGF loci proved the existence of a novel locus for autosomal dominant HGF and showed that this rare disorder is far more heterogeneous than previously expected.


Assuntos
Fibromatose Gengival/genética , Heterogeneidade Genética , Adolescente , Adulto , Idade de Início , Criança , Pré-Escolar , China , Mapeamento Cromossômico , Segregação de Cromossomos/genética , Cromossomos Humanos Par 11/genética , Cromossomos Humanos Par 2/genética , Cromossomos Humanos Par 5/genética , Feminino , Genes Dominantes/genética , Ligação Genética/genética , Genoma , Genótipo , Humanos , Lactente , Escore Lod , Masculino , Repetições de Microssatélites/genética , Linhagem , Penetrância , Adulto Jovem
12.
Arch Oral Biol ; 53(12): 1179-85, 2008 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-18760769

RESUMO

OBJECTIVES: To investigate the role of SecA in protein secretion, and to evaluate the effect of biofilm formation on protein secretion in Streptococcus mutans. DESIGN: S. mutans strains UA159 and GS-5 were used in this study. Cells grown in biofilm and planktonic conditions were observed using immunogold electron microscopy. The mRNA levels of ftf, gtfB, gtfC, gtfD, Pac and secA were analysed in different growth conditions using real-time quantitative polymerase chain reaction. The levels of wall proteins and whole-cell protein extracts were examined using Western blot analysis. RESULTS: A microdomain colocalised with SecA and virulence factors such as Pac (AgI/II) and glucosyltransferase (GTF) was observed. The mRNA level of secA was upregulated in the biofilm condition. The level of protein expression of SecA and wall protein levels of GTF, fructosyltransferase (FTF) and Pac (AgI/II) in the biofilm condition were significantly higher than in the planktonic condition. CONCLUSIONS: These data suggest that S. mutans utilises the Sec pathway to secrete virulence factor proteins such as Pac (AgI/II), GTF and FTF, and protein secretion occurred at a distinct microdomain. The level of SecA, the key factor in the Sec pathway, was influenced significantly by biofilm formation in S. mutans.


Assuntos
Aderência Bacteriana/fisiologia , Biofilmes/crescimento & desenvolvimento , Cárie Dentária/microbiologia , Saliva/microbiologia , Streptococcus mutans/fisiologia , Fatores de Virulência/metabolismo , Aderência Bacteriana/genética , Regulação Bacteriana da Expressão Gênica/genética , Ouro , Via Secretória/fisiologia , Streptococcus mutans/genética
13.
Arch Oral Biol ; 53(11): 1050-7, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18589399

RESUMO

OBJECTIVES: Hereditary gingival fibromatosis (HGF) is a rare benign disorder characterized by progressive fibrous overgrowth of the gingiva. The proliferation and expression of growth factors of HGF keratinocytes are abnormal. However, the exact role of keratinocytes in HGF pathogenesis is still unknown. The present study aimed to clarify the interactions between HGF keratinocytes and underlying fibroblasts in the pathogenesis of HGF. DESIGN: Gingival tissues, fibroblasts and keratinocytes from three Chinese HGF patients and three healthy subjects were collected. Histological analyses were performed by histochemical and immunohistochemical staining (Ki-67). Gingival fibroblasts were cocultured with gingival keratinocytes in an in vitro coculture system. The mRNA levels of type I collagen, MMP-1, MMP-3, and TIMP-1 were analysed in the cocultured gingival fibroblasts by reverse-transcription polymerase chain reaction (RT-PCR). The production of type I collagen and TIMP-1 was examined by ELISA. RESULTS: The number of Ki-67-positive keratinocytes in tissue sections from patients was higher than in those from controls. HGF fibroblasts cocultured with HGF keratinocytes showed an increased expression of type I collagen and TIMP-1. Transmission electron microscopy showed increased rough endoplasmic reticulum and ribosomes in cocultured HGF fibroblasts. CONCLUSIONS: These results suggest that HGF keratinocytes have an important role in HGF pathogenesis by inducing extracellular matrix (ECM) accumulation by fibroblasts.


Assuntos
Fibroblastos/metabolismo , Fibromatose Gengival/patologia , Queratinócitos/fisiologia , Adolescente , Adulto , Técnicas de Cultura de Células , Técnicas de Cocultura , Colágeno Tipo I/biossíntese , Colágeno Tipo I/genética , Feminino , Fibroblastos/ultraestrutura , Fibromatose Gengival/genética , Fibromatose Gengival/metabolismo , Expressão Gênica , Gengiva/metabolismo , Gengiva/ultraestrutura , Humanos , Masculino , Metaloproteinase 1 da Matriz/biossíntese , Metaloproteinase 1 da Matriz/genética , Metaloproteinase 3 da Matriz/biossíntese , Metaloproteinase 3 da Matriz/genética , Microscopia Eletrônica , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Inibidor Tecidual de Metaloproteinase-1/biossíntese , Inibidor Tecidual de Metaloproteinase-1/genética , Adulto Jovem
14.
Arch Oral Biol ; 52(12): 1209-14, 2007 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17825243

RESUMO

OBJECTIVES: Hereditary gingival fibromatosis (HGF) is characterized by excess accumulation of interstitial collagen. However, until now, there has been controversy about the mechanism of collagen accumulation in HGF gingivae. The present study aimed to clarify the pathogenic mechanisms potentially involved. DESIGN: Gingival fibroblasts from three Chinese HGF patients and three healthy subjects were cultured. Cell proliferation was assessed by MTT assay. The mRNA levels of type I collagen, MMP-1, MMP-3, TIMP-1, prolyl 4-hydroxylase (P4H)alpha(I), alpha(II), alpha(III) and P4Hbeta were analyzed in gingival fibroblasts by RT-PCR. The protein production of type I collagen and P4H was examined respectively by ELISA and Western blot. RESULTS: In culture, HGF gingival fibroblasts showed similar growth characteristics to fibroblasts isolated from control gingivae. The mRNA and protein levels of type I collagen and P4Halpha in HGF fibroblasts were higher than those in controls. There were no detected differences in mRNA expression levels of MMP-1, MMP-3, TIMP-1, P4Halpha(II), alpha(III) and P4Hbeta between HGF and control fibroblasts. CONCLUSIONS: These data suggest that increased collagen post-translational modification by P4H may be one mechanism by which increased collagen accumulation occurs in some forms of HGF.


Assuntos
Colágeno Tipo I/metabolismo , Fibroblastos/enzimologia , Fibromatose Gengival/enzimologia , Pró-Colágeno-Prolina Dioxigenase/metabolismo , Adolescente , Adulto , Feminino , Fibroblastos/metabolismo , Humanos , Masculino , Metaloproteinase 1 da Matriz/metabolismo , Metaloproteinase 3 da Matriz/metabolismo , RNA Mensageiro/análise , Inibidor Tecidual de Metaloproteinase-1/metabolismo
15.
Shanghai Kou Qiang Yi Xue ; 15(3): 285-9, 2006 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-16862364

RESUMO

PURPOSE: The purpose of this study was to compare the salivary immunoglobulin A antibody response to Streptococcus mutans in normal with in heat treated stress. METHODS: Clinical Streptococcus mutans strains were isolated from 20 volunteers, serotyped by biochemical test and PCR, and genotyped by AP-PCR. Unstimulated secretions from submandibular glands and sublingual glands were collected from volunteers by modified collectors. Each identified genotype was cultured in two groups: control group was grown in BHI broth at 37 degrees C. for 3 hours; stress group was incubated in BHI broth at 42 degrees C. for 3 hours. Analysis of SIgA activity to clinical genotype strains and reference strains in different group was detected by Western blot. RESULTS: There was no significant difference between stress group and control group,in spite that some bands had strong or weak intensity. Different genotypes of S.mutans could have different immunoblotting profile as for an individual. SIgA from different volunteers could have different immonoblotting profiles as to the same genotype strain. CONCLUSIONS: Although Streptococcus mutans can express heat shock proteins in stress, this study suggests these new proteins have no significant effect on the reaction of SIgA to Streptococcus mutans. Different genotype strains may have different proteins, and different immunoreactivity to host. Different hosts may have different immunoreactivities to one genotypes of S.mutans.


Assuntos
Resposta ao Choque Térmico , Imunoglobulina A Secretora/imunologia , Saliva/imunologia , Streptococcus mutans/imunologia , Western Blotting , Temperatura Alta , Humanos , Imunoglobulina A/imunologia
16.
Zhonghua Kou Qiang Yi Xue Za Zhi ; 41(1): 33-6, 2006 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-16620625

RESUMO

OBJECTIVE: To determine the physicochemical properties of the mutanase of Trichoderma harzianum isolated from China and to study the influence of mutanase on the adherence of oral Streptococci and the structure of oral biofilms. METHODS: Six fungal strains belonging to Trichoderma were tested for mutanase production in the same cultural condition, the strain producing the highest mutanase activity was studied further and the pH and temperature optimum of the enzyme was determined. The RT-PCR method was used to obtain the gene coding for mutanase and the product was cloned to pMD18-T simple vector for sequencing. Inhibition effects of mutanase on the adherence of Streptococcus sobrinus OMZ176, Streptococcus sobrinus 6715, Streptococcus mutans MT8148 were studied by adherence test. The optical sectioning of biofilms with or without mutanase supplementation were analyzed by confocal laser scanning microscopy (CLSM). RESULTS: The highest enzymatic activity was achieved by Trichoderma harzianum Th1, the maximum activity was at pH 5.5 and at 40 degrees C. The nucleotide sequence was 92% homology with that of a known gene coding a mutanase (GenBank accession No. AJ243799). The adherence of Streptococcus sobrinus OMZ176, Streptococcus sobrinus 6715, Streptococcus mutans MT8148 was significantly inhibited by mutanase. Compared with control, the biofilms with mutanase supplementation had lower height and sparser structure. CONCLUSIONS: The mutanase from Trichoderma harzianum Th1 can inhibit the adherence of oral Streptococci and had an influence on the structure of oral biofilms.


Assuntos
Aderência Bacteriana/efeitos dos fármacos , Biofilmes , Glicosídeo Hidrolases/química , Glicosídeo Hidrolases/fisiologia , Trichoderma/enzimologia , Streptococcus mutans/efeitos dos fármacos , Streptococcus sobrinus/efeitos dos fármacos , Trichoderma/patogenicidade
17.
Caries Res ; 39(6): 484-9, 2005 Nov-Dec.
Artigo em Inglês | MEDLINE | ID: mdl-16251793

RESUMO

The aim of the study was to examine the persistence of oral Streptococcus mutans in nasopharyngeal carcinoma patients after radiotherapy. Ten subjects, ranging in age from 20 to 67 years, participated. DMFT/DMFS, salivary level of mutans streptococci and oral health status were recorded. Pooled plaque samples were obtained from the cervical margins and the interproximal regions of all the teeth and the occlusal surfaces of the molars prior to, immediately after, 3 and 6 months after the completion of radiotherapy. At least 10 colonies of S. mutans were isolated from each subject and totally 645 isolates were genotyped by restriction endonuclease analysis. The results showed that the salivary level of S. mutans increased significantly with the reduction of salivary flow rate after radiotherapy. Each subject had at least 1 genotype of S. mutans isolated throughout the follow-up period. In 3 subjects who initially carried 2 or more genotypes, 1 or 2 genotypes of S. mutans could not be detected 3 months after treatment. Moreover, the genotypes that became undetectable were predominant bacteria in the first sampling. The result indicated that most S. mutans genotypes were persistent after radiotherapy but some genotypes that might not adapt to the alteration of oral environment became undetectable.


Assuntos
Neoplasias Nasofaríngeas/microbiologia , Saliva/efeitos da radiação , Streptococcus mutans/efeitos da radiação , Adulto , Idoso , Feminino , Humanos , Análise dos Mínimos Quadrados , Masculino , Pessoa de Meia-Idade , Neoplasias Nasofaríngeas/radioterapia , Saliva/microbiologia , Salivação/efeitos da radiação , Inquéritos e Questionários , Fatores de Tempo
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