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1	Estabilidade da oclusão tubular de diferentes agentes obliteradores para tratamento de hipersensibilidade dentinária Moura, Guilherme Faria 15 February 2016 (has links) Conselho Nacional de Desenvolvimento Científico e Tecnológico / In order to test the stability of dentinal tubules occlusion promoted by tubular front sealing agents to corrosive / abrasive challenges, the aim of our study is to evaluate the occlusion and stability of tubule-occluded products for the treatment of dentin hypersensitivity. Fifty sound tooth were randomly assigned in five groups (n=10), CL: Self Adhesive Sealant (Clinpro XT), TM: Calcium phosphate (Teethmate Desensitizer), GL: Glutaraldehyde (5% Gluma Desensitizer), NP: nano calcium phosphate (Nano P), 5% potassium oxalate (Painless). They were prepared to expose dentinal tubules of the cervical region, just below the cement enamel junction, submitted to treatment and later the challenges with hydrochloric acid and brushing, to simulate oral cavity of patients with gastroesophageal reflux disease (GERD). The analyzes were performed by counting, area and perimeter of the tubules, surface roughness, volume variation, height and angle vertical step, using the laser confocal microscope to evaluate the power of tubule-occluding agents and durability after the acids challenges and brushing. All criteria evaluated were submitted to appropriate statistical analysis. The effectiveness of the tested agents was evidenced in relation to time periods, all groups showed statistical differences between the initial and after treatment stages however only the CL, TM, GL groups did not achieve statistical differences between treatment and post challenge stages, proving stronger. When compared between groups, the CL was the only group that promoted complete obliteration of all dentinal tubules, maintained this obliteration after the challenges, and presents statistics differences in relation to the other groups in all variables. The use of desensitizing agents is effective in dentinal tubule occlusion. The CL, TM, GL agents were more resistant against biocorrosive-erosive challenges; the CL group was group that promoted better tubule obliteration. The surface roughness was higher for the CL group after the challenges. / O tratamento da hipersensibilidade dentinária é controverso, devido à falta de estudos que comprovam a efetividade e longa duração dos protocolos, assim como a sua estabilidade frente aos eventos de biocorrosão e abrasão que ocorrem na cavidade oral, principalmente em pacientes que sofrem de refluxo gastroesofágico. Esse tratamento é embasado nos princípios de vedamento do túbulo dentinário ou da dessensibilização das terminações nervosas periodontoblásticas dentro de cada túbulo. O objetivo do nosso trabalho foi avaliar o poder de obliteração e a durabilidade de produtos obliteradores para o tratamento da hipersensibilidade dentinária. Cinquenta (50) terceiros molares hígidos, foram preparados de modo a expor túbulos dentianários da região cervical, logo abaixo da junção cemento-esmalte e divididos aleatoriamente em cinco grupos CL: Selante autoadesivo (3M ESPE CLINPRO XT) (n=10), TM: Fosfato de cálcio (TEETHMATE DESENSITIZER KURARAY NORITAKE DENTAL INC.) (n=10), GL: Glutaraldeído 5% (GLUMA DESENSITIZER HERAEUS®) (n=10), NP: Fosfato de cálcio nano estruturado (NANO P FGM®) (n=10), PL: Oxalato de Potássio 5% (PAINLESS). Foram submetidos ao tratamento e posteriormente a desafios com ácido hidroclorídrico e escovação, para simular a cavidade oral, em pacientes portadores da doença do refluxo gastroesofágico. As análises foram realizadas por meio de contagem e perímetro dos túbulos, rugosidade de superfície e variação de volume, altura e degrau vertical utilizando microscópio confocal a laser, para avaliar a capacidade de obliteração dos produtos e a estabilidade após os desafios ácidos e de escovação. Os dados foram submetidos aos testes estatísticos apropriados (α=0,05). A eficácia dos agentes testados foi evidenciada em relação a períodos de tempo. Todos os grupos apresentaram diferença estatística entre os estágios iniciais e pós o tratamento (p<0,001), no entanto apenas o CL, TM, GL não apresentaram diferença estatística entre as etapas de tratamento e pós-desafio, mostrando maior resistência. Quando comparados entre si, o grupo CL promoveu maior obliteração dos túbulos 14 dentinários e manteve essa obliteração após os desafios, apresentando diferenças estatísticas, em relação aos outros grupos em todas as variáveis. O uso de agentes de dessensibilização é eficaz na oclusão dos túbulos dentinários. Os agentes CL, TM, GL foram mais resistentes a desafios biocorrosivos/erosivos, A rugosidade da superfície foi maior no grupo CL, após os desafios. / Mestre em Odontologia Refluxo gastroesofágico Dentina Hipersensibilidade dentinária Ánalise confocal Estabilidade de protocolos Obliteração de túbulos dentinários Tooth sensitivity Gastroesophageal reflux Confocal analysis Protocol durability dentinal tubule occlusion CNPQ::CIENCIAS DA SAUDE::ODONTOLOGIA
2	Development of Sensitive In Vitro Assays to Assess the Ocular Toxicity Potential of Chemicals and Ophthalmic Products McCanna, David January 2009 (has links) The utilization of in vitro tests with a tiered testing strategy for detection of mild ocular irritants can reduce the use of animals for testing, provide mechanistic data on toxic effects, and reduce the uncertainty associated with dose selection for clinical trials. The first section of this thesis describes how in vitro methods can be used to improve the prediction of the toxicity of chemicals and ophthalmic products. The proper utilization of in vitro methods can accurately predict toxic threshold levels and reduce animal use in product development. Sections two, three and four describe the development of new sensitive in vitro methods for predicting ocular toxicity. Maintaining the barrier function of the cornea is critical for the prevention of the penetration of infections microorganisms and irritating chemicals into the eye. Chapter 2 describes the development of a method for assessing the effects of chemicals on tight junctions using a human corneal epithelial and canine kidney epithelial cell line. In Chapter 3 a method that uses a primary organ culture for assessing single instillation and multiple instillation toxic effects is described. The ScanTox system was shown to be an ideal system to monitor the toxic effects over time as multiple readings can be taken of treated bovine lenses using the nondestructive method of assessing for the lens optical quality. Confirmations of toxic effects were made with the utilization of the viability dye alamarBlue. Chapter 4 describes the development of sensitive in vitro assays for detecting ocular toxicity by measuring the effects of chemicals on the mitochondrial integrity of bovine cornea, bovine lens epithelium and corneal epithelial cells, using fluorescent dyes. The goal of this research was to develop an in vitro test battery that can be used to accurately predict the ocular toxicity of new chemicals and ophthalmic formulations. By comparing the toxicity seen in vivo animals and humans with the toxicity response in these new in vitro methods, it was demonstrated that these in vitro methods can be utilized in a tiered testing strategy in the development of new chemicals and ophthalmic formulations. in vitro Alternative Methods alamarBlue rhodamine tight junctions bovine lens viability toxicity mitochondria mitochondria integrity benzalkonium chloride sodium dodecyl sulphate sodium lauryl sulfate scanning electron microscopy Draize Draize maximal average scores zonula occludens cosmetic directive PETA humane society animal league ICCVAM ECVAM JaCVAM BCOP bovine cornea ocular irritants ocular irritation ocular toxicity human corneal epithelial cells vitro confocal confocal microscopy metabolic activity optical quality reactive oxygen species contact lens contact lens care solutions barrier function microbial keratitis mulitipurpose solutions tight junction cell damage claudin ZO-1 occludin MDCK Madin Madin-Darby canine kidney cells cornea human cornea human corneal epithelium epithelial cell line human corneal epithelial cell line sodium fluorescein sodium fluorescein permeability fluorescein permeability ocular surface cell monolayer Araki-Sasaki cell physiology risk assessment safety assessment ScanTox scanning laser lens epithelium corneal cells in vitro model ophthalmic formulations multiple instillation back vertex animal testing rabbit testing alternatives to animal testing cultured bovine lens toxicity of chemicals irritation irritants laser scanner organ culture delayed toxicity toxins hydrogen peroxide cornea toxicity cornea toxicity models prediction of human toxicity no observable adverse effect level lowest observable adverse effect level NOAEL LOAEL toxicity thresholds safety factors cornea uncertainty factors preservatives disinfectants ophthalmic products preclinical preclinical testing epithelial barrier drug penetration clinical confocal microscopy animal rights rabbit cornea human cornea human clinical effects toxic animal rights activist sensitive measures toxic effect toxicity threshold agar overlay agar diffusion agar overlay method agar diffusion method cytochrome cytochrome c apoptosis necrotic apoptotic necrosis caspase rhodamine 123 resazuran resorufin cell death cell viability metabolic dye microsomal microsomal enzymes cytotoxicity cytotoxic cytotoxic effect MTT XTT WST-1 plasma membrane mitochondrial mitochondrial morphology ocular toxicity potential ocular toxicity human corneal epithelial cell line confocal analysis corneal epithelium cell fluorescence alamarBlue assay rhodamine dye animal welfare toxic injury degraded mitochondria epithelial monolayer disinfectants membrane integrity eye toxicity eye viability dye toxicity in humans human toxicity effects on the mitochondria mitochondrial toxicity in vitro battery in vitro test battery ophthalmic eye drop direct contact product safety cytotoxicity potential molecular molecular biology refine reduce replace sensitivity and relevance sensitivity relevance rabbit ocular irritation test product development cytotoxicity models cytotoxicity alternative methods replacements for animal testing three r's beagle tiered testing tiered testing strategy replacements animal testing mechanistic toxicity cornea mitochondria dose response threshold Vision Science and Biology
3	Development of Sensitive In Vitro Assays to Assess the Ocular Toxicity Potential of Chemicals and Ophthalmic Products McCanna, David January 2009 (has links) The utilization of in vitro tests with a tiered testing strategy for detection of mild ocular irritants can reduce the use of animals for testing, provide mechanistic data on toxic effects, and reduce the uncertainty associated with dose selection for clinical trials. The first section of this thesis describes how in vitro methods can be used to improve the prediction of the toxicity of chemicals and ophthalmic products. The proper utilization of in vitro methods can accurately predict toxic threshold levels and reduce animal use in product development. Sections two, three and four describe the development of new sensitive in vitro methods for predicting ocular toxicity. Maintaining the barrier function of the cornea is critical for the prevention of the penetration of infections microorganisms and irritating chemicals into the eye. Chapter 2 describes the development of a method for assessing the effects of chemicals on tight junctions using a human corneal epithelial and canine kidney epithelial cell line. In Chapter 3 a method that uses a primary organ culture for assessing single instillation and multiple instillation toxic effects is described. The ScanTox system was shown to be an ideal system to monitor the toxic effects over time as multiple readings can be taken of treated bovine lenses using the nondestructive method of assessing for the lens optical quality. Confirmations of toxic effects were made with the utilization of the viability dye alamarBlue. Chapter 4 describes the development of sensitive in vitro assays for detecting ocular toxicity by measuring the effects of chemicals on the mitochondrial integrity of bovine cornea, bovine lens epithelium and corneal epithelial cells, using fluorescent dyes. The goal of this research was to develop an in vitro test battery that can be used to accurately predict the ocular toxicity of new chemicals and ophthalmic formulations. By comparing the toxicity seen in vivo animals and humans with the toxicity response in these new in vitro methods, it was demonstrated that these in vitro methods can be utilized in a tiered testing strategy in the development of new chemicals and ophthalmic formulations. in vitro Alternative Methods alamarBlue rhodamine tight junctions bovine lens viability toxicity mitochondria mitochondria integrity benzalkonium chloride sodium dodecyl sulphate sodium lauryl sulfate scanning electron microscopy Draize Draize maximal average scores zonula occludens cosmetic directive PETA humane society animal league ICCVAM ECVAM JaCVAM BCOP bovine cornea ocular irritants ocular irritation ocular toxicity human corneal epithelial cells vitro confocal confocal microscopy metabolic activity optical quality reactive oxygen species contact lens contact lens care solutions barrier function microbial keratitis mulitipurpose solutions tight junction cell damage claudin ZO-1 occludin MDCK Madin Madin-Darby canine kidney cells cornea human cornea human corneal epithelium epithelial cell line human corneal epithelial cell line sodium fluorescein sodium fluorescein permeability fluorescein permeability ocular surface cell monolayer Araki-Sasaki cell physiology risk assessment safety assessment ScanTox scanning laser lens epithelium corneal cells in vitro model ophthalmic formulations multiple instillation back vertex animal testing rabbit testing alternatives to animal testing cultured bovine lens toxicity of chemicals irritation irritants laser scanner organ culture delayed toxicity toxins hydrogen peroxide cornea toxicity cornea toxicity models prediction of human toxicity no observable adverse effect level lowest observable adverse effect level NOAEL LOAEL toxicity thresholds safety factors cornea uncertainty factors preservatives disinfectants ophthalmic products preclinical preclinical testing epithelial barrier drug penetration clinical confocal microscopy animal rights rabbit cornea human cornea human clinical effects toxic animal rights activist sensitive measures toxic effect toxicity threshold agar overlay agar diffusion agar overlay method agar diffusion method cytochrome cytochrome c apoptosis necrotic apoptotic necrosis caspase rhodamine 123 resazuran resorufin cell death cell viability metabolic dye microsomal microsomal enzymes cytotoxicity cytotoxic cytotoxic effect MTT XTT WST-1 plasma membrane mitochondrial mitochondrial morphology ocular toxicity potential ocular toxicity human corneal epithelial cell line confocal analysis corneal epithelium cell fluorescence alamarBlue assay rhodamine dye animal welfare toxic injury degraded mitochondria epithelial monolayer disinfectants membrane integrity eye toxicity eye viability dye toxicity in humans human toxicity effects on the mitochondria mitochondrial toxicity in vitro battery in vitro test battery ophthalmic eye drop direct contact product safety cytotoxicity potential molecular molecular biology refine reduce replace sensitivity and relevance sensitivity relevance rabbit ocular irritation test product development cytotoxicity models cytotoxicity alternative methods replacements for animal testing three r's beagle tiered testing tiered testing strategy replacements animal testing mechanistic toxicity cornea mitochondria dose response threshold Vision Science and Biology

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