Orientador: Cláudia Valéria Seullner Brandão / Banca: Claudia Helena Pellizzon / Banca: Karina Preising Aptekmann / Resumo: A ceratoconjuntivite seca (CCS) é uma afecção de caráter inflamatório, comum em cães. O uso do ômega 3 e 6 no tratamento de humanos e coelhos com CCS tem sido pesquisado devido ao potencial efeito imunomodulador desses ácidos graxos, porém não há relatos do seu uso em cães com essa afecção. Objetivou-se avaliar o efeito dos óleos de linho e peixe, produtos ricos em ômega 3 e 6, administrados por via oral, na produção lacrimal e inflamação ocular de cães com CCS. Foram utilizados 45 cães, sendo 15 sem CCS, constituindo o grupo controle (GC), e 30 com CCS. Estes foram divididos de forma randomizada e cega em dois grupos: grupo tratado com óleo de linho (GL) e grupo tratado com óleo de peixe (GP); cada animal recebeu, por via oral, uma cápsula a cada 24 horas durante 60 dias, de acordo com o seu grupo. Exame clínico, teste lacrimal de Schirmer 1, teste de ruptura do filme lacrimal e teste de fluoresceína foram realizados antes do início do tratamento (M0) e aos 15 (M15), 30 (M30), 45 (M45) e 60 (M60) dias de tratamento. A citologia de impressão de superfície ocular foi realizada no M0, M30 e M60, enquanto a biópsia conjuntival para histopatologia foi obtida antes do tratamento e no momento final de avaliação. Houve diminuição significativa da hiperemia conjuntival nos cães de ambos os grupos tratados aos 60 dias. Na análise histopatológica, detectou-se diminuição da espessura epitelial no GL e GP, diminuição da queratinização no GL e diminuição do infiltrado de células inflamatórias no GP ao final do tratamento. Não houve diferença significativa nas demais avaliações. Conclui-se que os óleos de linho e peixe não aumentam a produção lacrimal até 60 dias do tratamento, porém melhoram a qualidade da superfície ocular. O óleo de peixe reduz o quadro inflamatório. Desta forma, o tratamento com óleo de linho ou peixe pode ser indicado como terapia adjuvante para cães com CCS / Abstract: The keratoconjunctivitis sicca (KCS) is inflammatory and a common disease in dogs. The use of omega 6 and 3 for treatment in humans and rabbits has been researched because of the potential immunomodulatory effect of these fatty acids, but there are no reports of its use in dogs with this condition. The aim of this study was to investigate the effect of linseed oil and fish oil, products rich in omega 3 and 6, administered orally on tear production and ocular inflammation in dogs with KCS. For this study we used 45 dogs: 15 without KCS, constituting the control group (CG) and 30 with KCS. A randomized, double-blind clinical study was performed in assigned to treatment orally with oil fish (FO) or linseed oil (LO), both once daily for 60 days. More 15 dogs with no KCS were evaluated for a control group (CG). Clinical examination of the eye, Schirmer tear tests I, tear film breakup time, fluorescein test were performed prior to treatment (M0) and at days 15 (M15), 30(M30), 45(M45) and 60(M60) after initiation of treatment. The impression cytology on ocular surface was performed at M0, M30 and M60. The conjunctival histopathological analysis was only performed prior to treatment and at day 60 after treatment. That was a significant decrease in conjunctival hyperemia in dogs of both treatments at day 60. The histopathological analysis revealed a decrease in the epithelial thickness both in FO and LO, a decrease on the degree of keratinization in LO and a decrease inflammatory cell infiltration in FO. There were no significant differences in the others evaluations in both groups. The analyzed results demonstrate that although both treatments administered orally do not increase tear production within 60 days of treatment, it improve the ocular surface. Thus, fish oil reduces eye inflammation. Therefore, treatment with linseed oil or fish may be indicated as adjunctive therapy for dogs with KCS / Mestre

MUC5B, une des 5 mucines gélifiantes chez les mammifères, constitue la matrice de gels de mucus et est responsable des propriétés visco-élastiques de ces gels. Cette macromolécule est sécrétée par des cellules caliciformes dont la densité reflète l'homéostasie des épithéliums sécrétoires.L'objectif principal de ce travail de thèse était la création et la caractérisation d'une souris transgénique afin de suivre facilement la production in vivo de Muc5b. Le second objectif était de montrer, chez des souris mucoviscidosiques CftrΔF508, qu'une nutrition enrichie en acides gras polyinsaturés à longues chaînes (n-3) module la production de la mucine Muc5b, confirmant des résultats précédemment publiés sur la régulation transcriptionnelle du gène Muc5b chez des souris de type sauvage par ce régime alimentaire.Une souris transgénique Muc5b-GFP a été créée par insertion d'une séquence codant la GFP (pour « Green Fluorescent Protein ») à la place du codon Stop de Muc5b. Le profil d'expression de Muc5b a été déterminé ex vivo sur tissus et chez la souris anesthésiée. Muc5b est produit dans l'oreille moyenne, l’œil, la cavité nasale, la trachée, la vésicule biliaire, le cervix, ainsi que dans le poumon embryonnaire à E12,5. Nous apportons la preuve de concept que Muc5b-GFP est un biomarqueur de la densité des cellules à mucus dans un modèle de syndrome de l’œil sec où nous mesurons, chez la souris anesthésiée, une diminution de la densité en cellules caliciformes qui est restaurée par application topique d'interleukine 13. La souris Muc5b-GFP est donc un modèle pré-clinique pour suivre la production de la mucine et la densité de cellules caliciformes. Dans le modèle murin CftrΔF508, nous montrons qu'une supplémentation à long terme en acides gras polyinsaturés à longues chaînes (n-3) n'influence pas le taux d'expression de Muc5b suite à une inflammation pulmonaire aiguë, mais améliore l'histologie pulmonaire et module la réponse inflammatoire. / MUC5B, one of the 5 gel-forming mucins in mammals, is part of the mucus gels and responsible of the visco-elastic properties of these gels. The density of goblet cells, which secrete this macromolecule, reflects the homeostasis of secretory epitheliums.The main goal of this thesis was the creation and characterization of a transgenic mouse to easily follow in vivo the production of Muc5b. The second goal was to show that, as previously reported by the laboratory in wild-type mice, a (n-3) long chain polyunsaturated fatty acid enriched diet modulates the production of Muc5b in the cystic fibrosis mouse model CftrΔF508.A Muc5b-GFP transgenic mouse was created by replacing the Stop codon of Muc5b with a GFP (Green Fluorescent Protein) coding sequence. Muc5b expression pattern was studied ex vivo in tissues and in anaesthetized mouse. Muc5b is produced in middle ear, eye, nasal cavity, trachea, gallbladder, cervix, and in embryonic lung at E12.5. As a proof-of-concept, we show that Muc5b-GFP is a biomarker of goblet cell density in a dry eye model. In anaesthetized mice, we measured a decreased goblet cell density after dry eye induction, which was restored by interleukin 13 topical application. Muc5b-GFP mouse is a preclinical model to follow the mucin production and goblet cell density. In the cystic fibrosis mouse model CftrΔF508, we show that a long term (n-3) long chain polyunsaturated fatty acid supplementation has no effect after an acute pulmonary inflammation on the Muc5b expression level, but induces an improvement of lung histology and a modulation of the inflammatory response.

Torra ögon beror på en obalans i ögats tårfilm och påverkar patientens livskvalité, eftersom tillståndet ger symtom i form av ögonsmärta, rinnande ögon och synnedsättning. Tillståndet är kroniskt, progressivt och kräver regelbunden behandling.  Syftet med denna studie var att undersöka patienters uppfattningar av att leva med torra ögon.  Metoden var en kvalitativ design med en fenomenografisk forskningsansats. Patienter från en ögonmottagning i Sydsverige blev enskilt intervjuade.  Resultatet visar på tre beskrivningskategorier: En förändrad tillvaro, Oro kring vad omgivningen tänker samt Att leva på hoppet. Informanterna uppfattar fysiska besvär i form av ögonsmärta och rinnande ögon, och upplever att besvären är obehagliga. Tillståndet uppfattas inverka negativt på livet och upplevs utgöra hinder för ett välfungerande vardagsliv. Informanterna uppfattar en oro kring vad omgivningen tänker om dem när besvären är synliga. De lever på hoppet om att hitta en lösning på besvären och vidtar olika åtgärder för att hantera tillståndets utmaningar.   Slutsatsen är att en ökad kunskap om patienters uppfattningar av att leva med torra ögon, kan leda till bättre stöd och omvårdnad i form av god information och personcentrerad omvårdnad. Det rekommenderas att torra ögon uppmärksammas mer inom omvårdnadsutbildning, och ytterligare forskning inom området rekommenderas. / Dry eyes are due to an imbalance in the tear film of the eye and affect the patient's quality of life, as the condition causes symptoms in form of eye pain, watery eyes and visual impairment. The condition is chronic, progressive and requires regular treatment.   The purpose of this phenomenographic study was to examine patient´s conceptions of living with dry eyes.   The method was a qualitative design with a phenomenographic research approach. Patients from an eye clinic in southern Sweden were individually interviewed.   The result shows three descriptive categories: A changed existence, Concern about what the environment thinks and Living on the hope. The informants perceive physical inconveniences such as eye pain and watery eyes, and experience that the inconveniences are unpleasant. The condition is perceived to have a negative impact on life and is perceived as a hindrance to a well-functioning everyday life. The informants perceive concerns about what the environment thinks of them when the problems are visible. They live on the hope of finding a solution to the problems and take various measures to deal with the challenges of the condition.  The conclusion is that an increased knowledge of patient´s conceptions of living with dry eyes, can lead to better support and nursing in form of good information and person-centered care. It is recommended that dry eyes be given more attention in nursing education, and further research in this area is recommended.

Dry eye syndrome has been associated with the lack of phospholipids in the tear film, leading to disruption of the tear film and subsequent irritation. Characterization of the transport and release of phospholipids from a silicone hydrogel contact lens is required to assess the possible use of these lenses for phospholipid delivery to increase patient comfort. This thesis examines the use of silicone hydrogel contact lenses as phospholipid delivery devices. Contact lenses of silicone hydrogel composition were loaded with varying amounts of radiolabeled 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) from a solution of n-propanol. These lenses were eluted at 35°C into artificial tear fluid (ATF) or ATFcontaining varying amounts of DMPC. The amount of DMPC loaded into a lens is a linear function of the time of exposure to the DMPC/propanol solution. The initial rate of elution into ATF appears to be diffusion controlled for at least 10 hrs and is proportional to the amount of DMPC loaded. The ease of loading and the controllable release of DMPC from silicone hydrogels present the possibility of using such lenses to counter eye discomfort caused by inherently low levels of phospholipid in tears. To reduce manufacturing steps and concern for residual n-propanol in the lens, it is beneficial to incorporate the DMPC into the monomer formulation and then photopolymerize the lens. Results showed that using this process, DMPC can be placed in the lens and then eluted at faster rates than when it was loaded from n-propanol.

In-situ chain extension of polymers used in the formulation of artificial tears and mild gelation are techniques to increase the residence time of eye drops on cornea. In-situ chain extension also helps to control the stability of ophthalmic emulsions both in the bottle and in the tear film. In this work, the interaction of hydrophobically modified guar and tear proteins as a method of polymer chain extension and mild gelation has been evaluated. Guar and its derivatives have been found to be very effective for ophthalmic applications. The ideal guar gelation agent is the one that turns on the gelation upon introduction onto the eye and that gelation chemistry is biocompatible and biodegradable. Controllable gelation is desirable to have relatively low viscosity eye drops for easy application and the drops form weak gels in the eye. One recent strategy to cure dry eye disease is to include emulsions in lubricant eye drops. The idea is to supplement the natural lipid layer on the exterior surface of the tear film. Formulating artificial tear emulsions is relatively complicated and must satisfy conflicting criteria. Emulsion droplets should be stable over the period of their shelf life without creaming or aggregate formation. On the other hand, in the tear film the emulsion droplets must cream fast enough and deposit onto the water/lipid film interface on the exterior surface of the tear film. Thus, the emulsion must be stable but not too stable. Initially, science-based design rules were proposed for the development of future generations of lubricant eye drops. The effect of guar molecular weight and concentration on emulsion stability was evaluated. According to the concentration-molecular weight plot, polymer solutions can be divided into stable and unstable regions. They are defined based on the critical flocculation concentration (CFC) and critical viscosity concentration (C*). Inverted QCM-D has been proposed as a simple and fast method to define the stability of oil in water emulsion systems. This technique is a promising alternative for time consuming conventional creaming experiments. Low molecular weight guar can be optimized to out-perform high molecular weight guars without the complications of formulating eye drops with high molecular weight polymers. Hydroxypropyl guar samples were oxidized and modified with linear alkyl amines to give a series of hydrophobically modified guars (MGuars). Lysozyme and human serum albumin (HSA), natural tear proteins, are able to extend the effective chain length of MGuar through polymer/protein complex formation. Hydrophobic modifications on guar enable efficient interaction with proteins, through their mutual hydrophobic characteristics. The interaction of proteins with various alkyl chain lengths, degrees of substitution and a range of molecular weights were examined. Binding and rheological measurements were employed to evaluate the interactions efficiency. Our results suggest that higher degrees of substitution and longer alkyl chain length give higher viscosity values. Lowering molecular weight allows for higher concentration, while keeping the initial viscosity constant. Higher viscosity was achieved as the chain extension occurred. The influence of hydrophobic modification and molecular weight variation on lubrication behavior of MGuars has also been determined. Hydrophobic modification enhanced the lubrication between hydrophobic surfaces. However, saturation of hydrophobes with protein abolished the lubricity. / Thesis / Doctor of Philosophy (PhD)

No description available.

Distúrbios na superfície da córnea e nas glândulas lacrimais acometem com frequência os indivíduos com diabetes mellitus. Atualmente não existe tratamento seguro e eficaz para feridas na córnea e o tratamento da síndrome do olho seco (SOS) é predominantemente sintomático. A administração tópica da insulina (INS) é uma estratégia promissora para tratar esses distúrbios, devido à presença de seus receptores na superfície ocular e na glândula lacrimal, e aos seus efeitos metabólicos e mitogênicos. No entanto, os fármacos aplicados topicamente na forma de solução são rapidamente drenados do olho, resultando em uma baixa biodisponibilidade local. O objetivo deste trabalho foi desenvolver formulações contendo INS e avaliar a sua influência no distúrbio das glândulas lacrimais e na regeneração da córnea em ratos diabéticos. Foram desenvolvidas quatro formulações contendo 1 UI/mL de INS: dispersão contendo insulina (DISP INS), dispersão contendo micropartículas quitosana/INS (DISP MP INS), gel termorreversível in situ com INS (Gel INS) e Gel contendo as micropartículas quitosana/INS (Gel MP INS). Também foram produzidas formulações \"brancas\", sem a veiculação do fármaco: dispersão contendo micropartículas sem insulina (DISP MP s/INS); gel termorreversível in situ sem insulina (Gel s/INS). As MP incorporadas nas formulações foram preparadas por spray drying e apresentaram tamanho de 4,0±0,1 ?m, morfologia adequada e grande quantidade de INS (77±6%). Todas as formulações apresentaram pH e osmolalidade compatíveis com o uso ocular. Durante o estudo in vivo, foi realizado o tratamento diário (15 dias) dos animais diabéticos, com as formulações (50 ?L) em ambos os olhos, exceto nos controles positivo (sem diabetes) e controle negativo (diabético não tratado). Todos os animais tratados com INS aumentaram a produção de fluido lacrimal, sendo que, ao término do tratamento, não foi observada diferença estatística entre o controle positivo e aqueles tratados com DISP INS e com Gel MP INS. A INS foi detectada na glândula lacrimal e no globo ocular dos animais tratados com DISP MP INS, Gel INS e Gel MP INS, sendo que a maior concentração de INS foi encontrada nos ratos tratados com Gel MP INS. Ao término do tratamento houve redução da glicemia dos animais tratados com a INS, o que pode sugerir um tratamento coadjuvante em pacientes diabéticos. Estudos de citologia de impressão mostraram que o Gel INS e Gel MP INS foram capazes de aumentar o número de células do epitélio da córnea e melhorar a relação núcleo/citoplasma em relação ao controle negativo. A histologia do globo ocular mostrou que essas duas formulações também melhoraram a espessura do epitélio da córnea, que foi semelhante a dos controles positivos. Conclui-se que a incorporação da INS nos géis e nas micropartículas desenvolvidas desempenhou um efeito positivo no tratamento da síndrome do olho seco e de lesões na córnea. / Disorders in the corneal surface and the lachrymal glands often affect individuals with diabetes mellitus. Nowadays there is no safe and effective treatment for wounds in the cornea and the treatment of dry eye syndrome (DES) is primarily symptomatic. Topical administration of insulin (INS) is a promising strategy for treating these disorders due to the presence of receptors on the ocular surface and lacrimal gland, besides its metabolic and mitogenic effects. However, the drugs applied topically in the form of a solution are quickly drained from the eye, resulting in a low local bioavailability. The aim of this study was to develop formulations containing INS and evaluate their influence on disorders of lacrimal glands and cornea healing in diabetic rats. Four formulations containing 1 IU/mL of INS were developed: dispersion containing insulin (DISP INS), dispersion containing chitosan microparticles/INS (DISP MP INS), in situ forming gel containing INS (Gel INS) and in situ forming gel containing microparticles chitosan/INS (Gel MP INS). \"White\" formulations were also produced without the drug: dispersion containing microparticles without insulin and in situ forming gel without insulin. MP incorporated in the formulations were prepared by spray drying and showed size of 4.0±0.1 ?m, proper morphology and high INS load (77±6%). All formulations exhibited pH and osmolarity compatible with the ocular use. In vivo studies were performed using diabetic rats that were treated daily (15 days) with instillation of the formulations (50 ?L) in both eyes, except in the positive controls (health rats) and negative control (untreated diabetic rats). All animals treated with INS showed an increase in the production of tear fluid. After treatment, no statistical difference was observed between the positive control and those treated with DISP INS and Gel MP INS. The INS was detected in the lacrimal gland of the animals treated with DISP MP INS, Gel INS and Gel MP INS, with the highest concentration of INS found in mice treated with Gel MP INS. After treatment there was a reduction in blood glucose of the animals treated with formulations containing INS, which suggests that developed formulations may be used as an adjuvant treatment in diabetic patients. Impression cytology studies showed that the Gel INS and Gel MP INS were able to increase the number of the corneal epithelium cells and improve nucleus/cytoplasm ratio compared to the negative control. The histology of the eyeball showed that these two formulations have improved the thickness of the corneal epithelium, which was similar to the positive controls. In conclusion, incorporation of INS in the gels and MP developed induced a positive effect in the treatment of dry eye syndrome and corneal wound.

A imunodeficiência comum variável (ICV) é uma imunodeficiência predominantemente humoral, com prejuízo na produção de imunoglobulinas. A maioria dos pacientes apresenta infecções recorrentes. No entanto, os pacientes com ICV podem apresentar comorbidades não infecciosas, dentre elas, alterações oftalmológicas. Manifestações oftalmológicas são muito pouco conhecidas em pacientes com ICV, porém alguns relatos de casos já foram descritos na literatura. Este trabalho teve como objetivos realizar avaliação oftalmológica completa dos pacientes com ICV dos ambulatórios de Imunologia do Hospital das Clínicas, procurando correlacionar as manifestações oftalmológicas encontradas com processos infecciosos e inflamatórios. Foram avaliados 16 pacientes, com idades entre 10 e 50 anos, dos quais seis apresentaram alterações oftalmológicas (três pacientes com síndrome do olho seco, um paciente com estrabismo, um paciente com pterígio e um paciente com retinopatia hipertensiva e espessamento da parede arteriolar). Entretanto, pterígio e estrabismo foram considerados alterações casuais, não sendo associados à ICV. Dois pacientes com olho seco fizeram avaliação da lágrima, com a finalidade de diagnóstico precoce de possível síndrome de Sjögren, mas a avaliação foi normal. Desta forma, são necessários mais estudos e com maior número de pacientes com finalidade de estabelecer marcadores laboratoriais preditivos para lesões oftalmológicas / The common variable immunodeficiency (CVID) is a predominantly humoral immunodeficiency, with impairment in the production of immunoglobulins. Most patients have recurrent infections. However, patients with CVID may have noninfectious comorbidities, including ophthalmological changes. Ophthalmologic manifestations are very little known in patients with CVID, but some case reports have already been described in the literature. The aim of this study was to perform a complete ophthalmologic evaluation of patients with CVID who are seeing in the Immunology Ambulatory of the Hospital das Clínicas, seeking to correlate the ophthalmological manifestations found with infectious and inflammatory processes. Sixteen patients were evaluated, between the ages of 10 and 50 years, six of which presented alterations (three patients with dry eye syndrome, one patient with strabismus, one patient with pterygium and one patient with hypertensive retinopathy and arteriolar wall thickening). However, pterygium and strabismus were considered casual alterations and were not associated with CVID. Two patients with dry eye made a tear evaluation, in view of the early diagnosis of Sjögren\'s Syndrome, but it was normal. Thus, additional studies with a larger patient sample are necessary to establish predictive laboratory markers for ophthalmological injuries

A imunodeficiência comum variável (ICV) é uma imunodeficiência predominantemente humoral, com prejuízo na produção de imunoglobulinas. A maioria dos pacientes apresenta infecções recorrentes. No entanto, os pacientes com ICV podem apresentar comorbidades não infecciosas, dentre elas, alterações oftalmológicas. Manifestações oftalmológicas são muito pouco conhecidas em pacientes com ICV, porém alguns relatos de casos já foram descritos na literatura. Este trabalho teve como objetivos realizar avaliação oftalmológica completa dos pacientes com ICV dos ambulatórios de Imunologia do Hospital das Clínicas, procurando correlacionar as manifestações oftalmológicas encontradas com processos infecciosos e inflamatórios. Foram avaliados 16 pacientes, com idades entre 10 e 50 anos, dos quais seis apresentaram alterações oftalmológicas (três pacientes com síndrome do olho seco, um paciente com estrabismo, um paciente com pterígio e um paciente com retinopatia hipertensiva e espessamento da parede arteriolar). Entretanto, pterígio e estrabismo foram considerados alterações casuais, não sendo associados à ICV. Dois pacientes com olho seco fizeram avaliação da lágrima, com a finalidade de diagnóstico precoce de possível síndrome de Sjögren, mas a avaliação foi normal. Desta forma, são necessários mais estudos e com maior número de pacientes com finalidade de estabelecer marcadores laboratoriais preditivos para lesões oftalmológicas / The common variable immunodeficiency (CVID) is a predominantly humoral immunodeficiency, with impairment in the production of immunoglobulins. Most patients have recurrent infections. However, patients with CVID may have noninfectious comorbidities, including ophthalmological changes. Ophthalmologic manifestations are very little known in patients with CVID, but some case reports have already been described in the literature. The aim of this study was to perform a complete ophthalmologic evaluation of patients with CVID who are seeing in the Immunology Ambulatory of the Hospital das Clínicas, seeking to correlate the ophthalmological manifestations found with infectious and inflammatory processes. Sixteen patients were evaluated, between the ages of 10 and 50 years, six of which presented alterations (three patients with dry eye syndrome, one patient with strabismus, one patient with pterygium and one patient with hypertensive retinopathy and arteriolar wall thickening). However, pterygium and strabismus were considered casual alterations and were not associated with CVID. Two patients with dry eye made a tear evaluation, in view of the early diagnosis of Sjögren\'s Syndrome, but it was normal. Thus, additional studies with a larger patient sample are necessary to establish predictive laboratory markers for ophthalmological injuries

Introdução: As queimaduras alcalinas da córnea fazem parte dos ferimentos mais devastadores para o olho. Objetivo: Os objetivos foram investigar o efeito aditivo do olho seco na atividade de proteases na superfície ocular e complicações corneanas apos lesão ocular alcalina e também investigar a eficácia da terapia anti-inflamatória controlar este processo. Métodos: Um modelo combinado (CM) de olho seco e queimadura alcalina unilateral foi usada. Resumidamente, camundongos C57BL /6 foram submetidos à queimadura alcalina unilateral (AB) com ou sem olho seco concomitante por 2 ou 5 dias. Um grupo separado de animais foram submetidos a ambos modelos (AB e olho seco) foram tratados topicamente com a Dexametasona, ou Doxiciclina ou colírio controle de solução salina balanceada (BSS). Os camundongos foram observados diariamente para verificar o aparecimento de perfuração da córnea. Córneas inteiras foram colhidas e homogeneizadas para extração de RNA. PCR quantitativo em tempo real foi realizada para medir a expressão de citocinas inflamatórias, metaloproteinases de matriz (MMP). Ativdade da MMP-9, atividade da gelatinase e da atividade da mieloperoxidase (MPO) foram avaliados em córneas homogeneizadas. A presença da infiltração de neutrófilos foi avaliada por imunohistoquímica e citometria de fluxo. Resultados: Os olhos submetidos ao modelo combinado de AB e olho seco (CM) tiveram 20% de taxa de perfuração estéril da córnea 1 dia após a lesão inicial; que aumentou para 35% em 5 dias. Houve um atraso no fechamento da ferida e aumento de opacidade residual da córnea. Aumento dos níveis de IL-1, IL-6, e as MMPs 1, -3, -8, -9, 13 e CXCL1, foram encontrados após 2 dias no CM comparando com córneas AB. Um aumento da imunorreatividade da MMP-1, -3, -9 e -13 e atividade gelatinolítica da MMP-9 foram observadas em comparação com córneas do grupo CM comparado com AB. O aumento da infiltração de neutrófilos e a atividade da mieloperoxidase foi observado no grupo CM comparando-se com córneas do grupo AB após dois dias da lesão inicial. Não foram observadas perfurações nas córneas tratadas com Dexametasona. Nos olhos tratados com Doxiciclina, 100% do fechamento da ferida pós-lesão no dia 2 e pontuação significantemente menor na escala de opacidade da córnea em relação ao BSS também foram observadas nos dias 4 e 5. Córneas tratadas com Dexametasona apresentaram a menor pontuação de opacidade da córnea. Tratamento com Dexametasona diminuiu significativamente os níveis de mRNA da IL-1, IL-6, e MMP-1, -9, -13, e o TIMP-1 depois de 2 dias, e aumentou os níveis de MMP-8, enquanto que o tratamento com Doxiciclina diminuiu significativamente IL-1, IL-6, MMP-8, -9 e, em comparação com córneas tratadas com BSS. A diminuição da imunorreatividade da MMP-1, -9 e -13 e atividade gelatinolítica foram vistos em córneas tratadas com Doxiciclina e Dexametasona em comparação com o veículo BSS. O aumento da infiltração de neutrófilos e a atividade da mieloperoxidase foi observado no grupo BSS comparação com o grupo Dexametasona 2 dias pós-lesão. Conclusões: O olho seco ambiental piora o resultado da queimadura ocular alcalina, criando uma tempestade de citocinas e proteases, aumentando o risco de perfuração corneana. Entretanto, o tratamento inicial com terapia anti-inflamatória é muito eficaz na preservação da transparência corneana e facilita a cicatrização de feridas, enquanto controla a produção de MMP e a migração de neutrófilos. / Introduction: Alkali burns to the cornea are among the most devastating injuries to the eye. Purpose: To evaluate the effects of dry eye on ocular surface protease activity and sight threatening corneal complications following ocular surface chemical injury and also to investigate the efficacy of anti-inflammatory therapy controlling this. Methods: A combined model (CM) of unilateral alkali burn and dry eye was used. Briefly, C57BL/6 mice were subjected to unilateral alkali burn (AB) with or without concomitant dry eye for 2 or 5 days. A separate group of mice subjected to both AB and dry eye were topically treated with Dexamethasone (Dex), Doxycycline (Doxy) or saline control (BSS). Mice were observed daily for appearance of corneal perforation. Whole corneas were harvested and lysed for RNA extraction. Quantitative real time PCR was performed to measure expression of inflammation cytokines, matrix metalloproteinases (MMP). MMP-9 activity, gelatinase activity and myeloperoxidase (MPO) activity were evaluated in corneal lysates. Presence of infiltrating neutrophils was evaluated by immunohistochemistry and flow cytometry. Results: Eyes subjected to the combined model of AB and dry eye (CM) had 20% sterile corneal perforation rate as soon as 1 day after the initial injury, which increased to 35% by 5 days, delayed wound closure and increased corneal opacity. Increased levels of IL- 1, IL-6, and MMPs 1,-3,-8,-9, 13, and CXCL1 transcripts were found after 2 days in CM compared to AB corneas. Increased MMP-1, -3,-9 and -13 immunoreactivity and gelatinolytic activity were seen in CM corneas compared to AB. Increased neutrophil infiltration and MPO activity was noted in the CM group compared to AB 2 days post injury. No perforations were observed in the Dex treated corneas. Doxy treated eyes had 100% of wound closure 2D post-injury, and significant lower corneal opacity scores at days 4 and 5 compared to BSS. Dex-treated corneas showed the lowest corneal opacity score. Dex treatment significantly decreased mRNA levels of IL-1, IL-6, and MMPs -1,-9, -13, and TIMP-1 after 2 days with increased levels of MMP-8, while Doxy treatment significantly decreased IL-1, IL-6, MMP-8, and -9, compared to BSS-treated corneas. Decreased MMPs -1,-9 and -13 immunoreactivity and gelatinolytic activity were seen in corneas treated with Doxy and Dex compared to BSS vehicle. Increased neutrophil infiltration and MPO activity was noted in the BSS group compared to Dex 2 D post-injury. Conclusions: Desiccating stress worsens outcome of ocular alkali burn, creating a cytokine and protease storm with greater neutrophil infiltration, increasing the risk of corneal perforation. However, early treatment with anti-inflammatory therapy is very efficacious in preserving corneal clarity and facilitating wound healing, while controlling MMP production and migration of neutrophils.