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1.
PLoS One ; 15(6): e0235067, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32559226

RESUMO

The aim of this study was to investigate whether and how the biological media which are in contact with silicone oil play a role in the silicone emulsification process. Commercially available Oxane 1300 silicone oil and potential hydrophilic phases of the emulsions in the eye (porcine aqueous humor, porcine vitreous and balanced salt solution) were investigated separately and in a mixture or emulsions by means of surface tension, rheological, zeta potential measurements and microscopic investigation. The surface tension of biological media (vitreous and aqueous humor) was significantly lower than that of non-biological media, especially in the case of aqueous humor, which indicates a remarkable emulsification tendency with these phases. The biological media are able to form both oil-in-water and water-in-oil emulsions, which can be observed in the clinical practice as well. It was established that the vitreous has a more expressed emulsification ability compared with the aqueous humor because smaller and more stable droplets can form with silicon oil when the vitreous is still there. It can be concluded that the vitreous has a higher impact on emulsification than the aqueous medium, which can predict that the vitreous remaining after vitrectomy has a key role in emulsion formation in the eye with silicone oil endotamponade.


Assuntos
Humor Aquoso/química , Soluções Isotônicas/química , Óleos de Silicone/química , Corpo Vítreo/química , Animais , Emulsões , Reologia , Tensão Superficial , Suínos , Vitrectomia/métodos
2.
BMC Ophthalmol ; 18(1): 298, 2018 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-30428856

RESUMO

BACKGROUND: We aimed to analyze the differences in the prevalence of outer retinal tubulation (ORT) in neovascular age-related macular degeneration (AMD) treated with anti-vascular endothelial growth factor (anti-VEGF) agents, either aflibercept or ranibizumab. Our further aim was to examine the changes in the frequency of injections of ranibizumab before and after ORT appearance. METHODS: Two hundred thirty six eyes of 230 patients were included in the study (184 eyes treated with ranibizumab by pro re nata regimen (PRN), 52 eyes with aflibercept bimonthly) and followed for 6-24 months. Using optical coherence tomography (OCT), the first appearance of ORT was documented, and fixed time point evaluations were also made every six months to determine the existence of ORT. The number of injections, the presence or absence of subretinal hyperreflective material (SHRM) at treatment initiation and visual acuity were also noted. RESULTS: The survival analysis with Cox proportional hazard model showed no significant difference between the ranibizumab and aflibercept groups in relation to the development of ORT (p = 0.79, hazard ratio 0.92). In the PRN treated ranibizumab group the number of injections showed significant decrease after ORT development (p = 0.004). When SHRM was present at treatment initiation the chance of developing ORT was 2.75 and 11.14 times higher in the ranibizumab and aflibercept groups, respectively. CONCLUSIONS: The prevalence of ORT increased over time independently from the chosen anti-VEGF drug. Our results suggest that upon the appearance of ORT a decrease in retreatments can be expected.


Assuntos
Inibidores da Angiogênese/uso terapêutico , Degeneração Macular/tratamento farmacológico , Ranibizumab/uso terapêutico , Receptores de Fatores de Crescimento do Endotélio Vascular/uso terapêutico , Proteínas Recombinantes de Fusão/uso terapêutico , Retina/patologia , Idoso , Idoso de 80 Anos ou mais , Biomarcadores , Feminino , Humanos , Degeneração Macular/patologia , Masculino , Pessoa de Meia-Idade , Prevalência , Retratamento , Estudos Retrospectivos , Acuidade Visual
3.
Histol Histopathol ; 33(5): 455-462, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-28872183

RESUMO

INTRODUCTION: Multilamellar bodies (MLBs) are concentric cytoplasmic membranes which form through an autophagy-dependent mechanism. In the cornea, the presence of MLBs is associated with Schnyder corneal dystrophy (SCD). Ex vivo 3D modelling of the corneal stroma and SCD can help study pathogenesis and resolution of the disorder. METHODS: Corneal stroma explants were isolated from cadavers and cultivated long-term for more than 3 months to achieve spontaneous 3D outgrowth of corneal stroma-derived mesenchymal stem-like cells (CSMSCs). The 3D tissues were then examined by transmission electron microscopy (TEM) for presence of MLBs, and by immunofluorescent labelling against markers for autophagy (p62, LC3). Autophagy was induced by classical serum starvation or rapamycin (RAP) treatment (50 nM), and inhibited by the autophagy inhibitor 3-methyladenine (3-MA, 10 mM) for 24 hours. RESULTS: CSMSCs can form spontaneously 3D outgrowths over a 3-4 weeks period, depositing their own extracellular matrix containing collagen I. TEM confirmed the presence of MLBs in the long-term (>3 months) 3D cultures, which became more abundant under starvation and RAP treatment, and decreased in number under autophagy inhibition with 3-MA. The presence of autophagy and its disappearance could be confirmed by an inversely related increase and decrease in the expression of LC3 and p62, respectively. CONCLUSIONS: MLB formation in long-standing CSMSC cultures could serve as a potential ex vivo model for studying corneal stroma diseases, including SCD. Inhibition of autophagy can decrease the formation of MLBs, which may lead to a novel treatment of the disease in the future.


Assuntos
Autofagia , Distrofias Hereditárias da Córnea/patologia , Substância Própria/patologia , Adenina/análogos & derivados , Adenina/farmacologia , Adulto , Idoso , Autofagia/efeitos dos fármacos , Cadáver , Células Cultivadas , Córnea/metabolismo , Distrofias Hereditárias da Córnea/fisiopatologia , Substância Própria/fisiopatologia , Substância Própria/ultraestrutura , Matriz Extracelular/metabolismo , Matriz Extracelular/ultraestrutura , Imunofluorescência , Humanos , Imageamento Tridimensional , Corpos de Inclusão/patologia , Corpos de Inclusão/ultraestrutura , Células-Tronco Mesenquimais , Microscopia Eletrônica de Transmissão , Pessoa de Meia-Idade , Modelos Anatômicos
4.
Orv Hetil ; 158(39): 1523-1527, 2017 Oct.
Artigo em Húngaro | MEDLINE | ID: mdl-28942662

RESUMO

Measles, caused by the Morbilli virus, is a highly (about 95 %) contagious disease affecting primarily children, but without proper immunisation, adults can also be infected. The leading symptoms of the disease are high fever that presents after an incubation period of 9-10 days and the red rash that begins several days after the fever starts. Beyond specific generalized symptoms, measles may have ocular symptoms. The most commonly occurring conjunctivitis, the so-called "red eye symptom", is not characteristic only for measles infection, however, by taking the generalized symptoms it can suggest the diagnosis at the beginning of the disease. Conjunctivitis of varying severity is noticed in the half of the cases without using ophthalmological instrumentation. Using ophthalmological instrumentation, the mild forms of conjunctivitis can be diagnosed, by meticulous ophthalmological examination, further eye diseases can be discovered. The viral conjunctivitis can progress to keratitis and bacterial superinfection can occur. If the infection presents in childhood it can affect the posterior segment. The fight against measles is very effective in Hungary since the vaccination has been introduced, and the lack of vaccination is also the primary cause of the risk to the disease. In the diagnosis, symptomatic treatment of the disease and the curbing of possible mass infections, the practicing physician (general practitioner) has a key role. The correct care of the infected patient in Hungary is provided by a methodological letter, professional information and legal guides. Orv Hetil. 2017; 158(39): 1523-1527.


Assuntos
Conjuntivite/diagnóstico , Conjuntivite/etiologia , Sarampo/diagnóstico , Sarampo/terapia , Adulto , Antibacterianos/uso terapêutico , Criança , Humanos , Hungria , Sarampo/complicações , Oftalmologia
5.
Orv Hetil ; 158(18): 701-705, 2017 May.
Artigo em Húngaro | MEDLINE | ID: mdl-28468537

RESUMO

INTRODUCTION: CO2 laser- assisted sclerectomy surgery (CLASS) can be used for the surgical treatment of open-angle glaucoma. AIM: To introduce our results with CLASS. METHOD: We performed 21 CLASS operations using OT-134-IOPtiMate (IOPtima Ltd, Ramat-Gan, Israel). Patients were examined on the 1st day, and in the 1st, 3rd, 6th, 9th and 12th months postoperatively. We evaluated intraocular pressure (IOP), antiglaucomatous medication-use, visual acuity, complications. RESULTS: Mean age was 65.6 yrs. Complete success (no hypotensive medication required to target IOP) was achieved in 61.1% (18 patients) at 6 months, whereas in 50% (10 patients) at 12 months. Qualified success (hypotensive medication required to target IOP) was achieved in 72.2% and in 70%, preoperative mean IOP was 29.2 ± 9.4 Hgmm, which falled to 17.7 ± 4.9 Hgmm and 17.3 ± 4.3 Hgmm, respectively. Antiglaucomatous medication use falled significantly from 2.90 ± 0.83 to 2.05 ± 1.46. Apart from 1 macroperforation, no serious complication occurred. CONCLUSIONS: With CLASS it is possible to effectively lower intraocular pressure in open-angle glaucoma. Orv Hetil. 2017; 158(18): 701-705.


Assuntos
Glaucoma de Ângulo Aberto/cirurgia , Lasers de Gás/uso terapêutico , Esclera/cirurgia , Esclerostomia/métodos , Adulto , Idoso , Feminino , Glaucoma de Ângulo Aberto/fisiopatologia , Humanos , Pressão Intraocular/fisiologia , Masculino , Pessoa de Meia-Idade , Procedimentos Cirúrgicos Oftalmológicos/métodos , Resultado do Tratamento , Acuidade Visual/fisiologia
6.
Cont Lens Anterior Eye ; 40(5): 283-292, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28550976

RESUMO

PURPOSE: Development of ex vivo model to study pathogenesis, inflammation and treatment modalities for pterygium. METHODS: Pterygium obtained from surgery was cultivated (3 months). Gravitational attachment method using viscoelastic facilitated adherence of graft and outgrowing cells. Medium contained serum as the only growth supplement with no use of scaffolds. Surface profiling of the multi-layered cells for hematopoietic- and mesenchymal stem cell markers was performed. Examination of cells by immunohistochemistry using pluripotency, oxidative stress, stemness, migration and proliferation, epithelial and secretory markers was performed. The effect of anti-proliferative agent Mitomycin C upon secretion of pro-inflammatory cytokines IL-6 and IL-8 was assessed. RESULTS: Cells showed high expression of migration- (CXCR4), secretory- (MUC1, MUC4) and oxidative damage- (8-OHdG) markers, and low expression of hypoxia- (HIF-1α) and proliferation- (Ki-67) markers. Moderate and low expression of the pluripotency markers (Vimentin and ΔNp63) was present, respectively, while the putative markers of stemness (Sox2, Oct4, ABCG-2) and epithelial cell markers- (CK19, CK8-18) were weak. The surface marker profile of the outgrowing cells revealed high expression of the hematopoietic marker CD47, mesenchymal markers CD90 and CD73, minor or less positivity for the hematopoietic marker CD34, mesenchymal marker CD105, progenitor marker CD117 and attachment protein markers while low levels of IL-6 and IL-8 secretion ex vivo, were inhibited upon Mitomycin C treatment. CONCLUSION: Ex vivo tissue engineered pterygium consists of a mixture of cells of different lineage origin, suitable for use as a disease model for studying pathogenesis ex vivo, while opening possibilities for new treatment and prevention modalities.


Assuntos
Modelos Biológicos , Pterígio/patologia , Alquilantes/farmacologia , Biomarcadores/metabolismo , Movimento Celular , Proliferação de Células , Ensaio de Imunoadsorção Enzimática , Técnica Indireta de Fluorescência para Anticorpo , Células-Tronco Hematopoéticas/metabolismo , Humanos , Imunofenotipagem , Células-Tronco Mesenquimais/metabolismo , Mitomicina/farmacologia , Técnicas de Cultura de Órgãos , Pterígio/metabolismo , Pterígio/terapia
7.
Invest Ophthalmol Vis Sci ; 57(8): 3828-35, 2016 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-27438543

RESUMO

PURPOSE: We recently reported that isolated duct segments from rabbit lacrimal gland (LG) were able to secrete fluid in response to secretagogues, which were blocked completely by bumetanide. This suggests the functional involvement of Na+-K+-2Cl- cotransporter (NKCC1) in ductal fluid secretion. Therefore, the aim of this study was to investigate the activity profile of NKCC1 in isolated rabbit LG duct segments. METHODS: Interlobular ducts were isolated from fresh rabbit LG tissue. Microfluorometry with the ammonium (NH4+)-pulse technique was used to elicit pH changes in duct cells, and the rate of bumetanide-sensitive cytosolic acidification after addition of NH4+ was used to quantify the activity of NKCC1. RESULTS: While basal activity of NKCC1 was undetectable, low cytosolic chloride (Cl-) level and hyperosmotic challenge (390 mOsm) were able to increase the activity of NKCC1. Carbachol (100 µM) had no significant effect on NKCC1 activity. Elevation of cytosolic calcium (Ca2+) level with Ca2+-ionophore (A 23187, 1 µM) did not cause any alteration in the activity of the cotransporter while direct activation of protein kinase C (phorbol myristate acetate, 100 nM) increased its activity slightly but in a significant manner. Addition of either forskolin (10 µM), cell-permeable cAMP analogue (8-bromo cAMP, 100 µM) or vasoactive intestinal peptide (200 nM) resulted in a significant increase in the activity of NKCC1. CONCLUSIONS: These results highlight the functional involvement of NKCC1 in LG duct secretion. These findings may facilitate our understanding of LG function and may contribute to the development of targeted pharmacologic interventions in case of dry eye disease.


Assuntos
Aparelho Lacrimal/metabolismo , Membro 2 da Família 12 de Carreador de Soluto/fisiologia , Análise de Variância , Animais , Carbacol/farmacologia , Carcinógenos/farmacologia , Colforsina/farmacologia , GMP Cíclico/análogos & derivados , GMP Cíclico/farmacologia , Síndromes do Olho Seco/etiologia , Síndromes do Olho Seco/metabolismo , Concentração de Íons de Hidrogênio , Técnicas In Vitro , Masculino , Soluções Oftálmicas/farmacologia , Concentração Osmolar , Coelhos , Membro 2 da Família 12 de Carreador de Soluto/metabolismo , Lágrimas/metabolismo , Acetato de Tetradecanoilforbol/farmacologia , Peptídeo Intestinal Vasoativo/farmacologia
8.
Sci Rep ; 6: 26227, 2016 05 19.
Artigo em Inglês | MEDLINE | ID: mdl-27195722

RESUMO

Corneal tissue regeneration is of crucial importance for maintaining normal vision. We aimed to isolate and cultivate human corneal stroma-derived mesenchymal stem-like cells (CSMSCs) from the central part of cadaver corneas and study their phenotype, multipotency, role in immunity and wound healing. The isolated cells grew as monolayers in vitro, expressed mesenchymal- and stemness-related surface markers (CD73, CD90, CD105, CD140b), and were negative for hematopoietic markers as determined by flow cytometry. CSMSCs were able to differentiate in vitro into fat, bone and cartilage. Their gene expression profile was closer to bone marrow-derived MSCs (BMMSCs) than to limbal epithelial stem cells (LESC) as determined by high-throughput screening. The immunosuppressive properties of CSMSCs were confirmed by a mixed lymphocyte reaction (MLR), while they could inhibit proliferation of activated immune cells. Treatment of CSMSCs by pro-inflammatory cytokines and toll-like receptor ligands significantly increased the secreted interleukin-6 (IL-6), interleukin-8 (IL-8) and C-X-C motif chemokine 10 (CXCL-10) levels, as well as the cell surface adhesion molecules. CSMSCs were capable of closing a wound in vitro under different stimuli. These cells thus contribute to corneal tissue homeostasis and play an immunomodulatory and regenerative role with possible implications in future cell therapies for treating sight-threatening corneal diseases.


Assuntos
Córnea/imunologia , Córnea/fisiologia , Substância Própria/fisiologia , Células-Tronco Mesenquimais/fisiologia , Cicatrização , Idoso , Idoso de 80 Anos ou mais , Antígenos de Superfície/análise , Diferenciação Celular , Células Cultivadas , Citocinas/metabolismo , Feminino , Citometria de Fluxo , Perfilação da Expressão Gênica , Ensaios de Triagem em Larga Escala , Humanos , Masculino , Células-Tronco Mesenquimais/química , Pessoa de Meia-Idade , Modelos Biológicos , Regeneração
9.
J Diabetes Res ; 2016: 4529824, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-28078306

RESUMO

Introduction. Diabetic retinopathy (DR) is a sight-threatening complication of diabetes. Telemedicine tools can prevent blindness. We aimed to investigate the patients' satisfaction when using such tools (fundus camera examination) and the effect of demographic and socioeconomic factors on participation in screening. Methods. Pilot study involving fundus camera screening and self-administered questionnaire on participants' experience during fundus examination (comfort, reliability, and future interest in participation), as well as demographic and socioeconomic factors was performed on 89 patients with known diabetes in Csongrád County, a southeastern region of Hungary. Results. Thirty percent of the patients had never participated in any ophthalmological screening, while 25.7% had DR of some grade based upon a standard fundus camera examination and UK-based DR grading protocol (Spectra™ software). Large majority of the patients were satisfied with the screening and found it reliable and acceptable to undertake examination under pupil dilation; 67.3% were willing to undergo nonmydriatic fundus camera examination again. There was a statistically significant relationship between economic activity, education and marital status, and future interest in participation. Discussion. Participants found digital retinal screening to be reliable and satisfactory. Telemedicine can be a strong tool, supporting eye care professionals and allowing for faster and more comfortable DR screening.


Assuntos
Retinopatia Diabética/diagnóstico , Oftalmologia/instrumentação , Telemedicina/instrumentação , Telemedicina/métodos , Idoso , Feminino , Fundo de Olho , Humanos , Hungria , Masculino , Programas de Rastreamento/métodos , Pessoa de Meia-Idade , Oftalmologia/métodos , Satisfação do Paciente , Projetos Piloto , Reprodutibilidade dos Testes , Classe Social , Software , Inquéritos e Questionários
10.
PLoS One ; 10(11): e0143053, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26580800

RESUMO

Long-term cultures of cornea limbal epithelial stem cells (LESCs) were developed and characterized for future tissue engineering and clinical applications. The limbal tissue explants were cultivated and expanded for more than 3 months in medium containing serum as the only growth supplement and without use of scaffolds. Viable 3D cell outgrowth from the explants was observed within 4 weeks of cultivation. The outgrowing cells were examined by immunofluorescent staining for putative markers of stemness (ABCG2, CK15, CK19 and Vimentin), proliferation (p63α, Ki-67), limbal basal epithelial cells (CK8/18) and differentiated cornea epithelial cells (CK3 and CK12). Morphological and immunostaining analyses revealed that long-term culturing can form stratified 3D tissue layers with a clear extracellular matrix deposition and organization (collagen I, IV and V). The LESCs showed robust expression of p63α, ABCG2, and their surface marker fingerprint (CD117/c-kit, CXCR4, CD146/MCAM, CD166/ALCAM) changed over time compared to short-term LESC cultures. Overall, we provide a model for generating stem cell-rich, long-standing 3D cultures from LESCs which can be used for further research purposes and clinical transplantation.


Assuntos
Células Epiteliais/citologia , Epitélio Corneano/citologia , Limbo da Córnea/citologia , Cultura Primária de Células/métodos , Células-Tronco/citologia , Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP , Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Antígenos CD/genética , Antígenos CD/metabolismo , Autopsia , Biomarcadores/metabolismo , Diferenciação Celular , Proliferação de Células , Colágeno/genética , Colágeno/metabolismo , Meios de Cultura , Células Epiteliais/metabolismo , Epitélio Corneano/metabolismo , Expressão Gênica , Humanos , Queratinas/genética , Queratinas/metabolismo , Antígeno Ki-67/genética , Antígeno Ki-67/metabolismo , Limbo da Córnea/metabolismo , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Células-Tronco/metabolismo , Engenharia Tecidual , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Proteínas Supressoras de Tumor/genética , Proteínas Supressoras de Tumor/metabolismo , Vimentina/genética , Vimentina/metabolismo
11.
Int J Pharm ; 494(1): 321-8, 2015 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-26319587

RESUMO

Hyaluronic acid (HA) and its derivatives play important roles in many fields of therapy, such as arthritis treatment, plastic surgery, dermatology, otology, ophthalmology, etc. With a view to increase the beneficial properties of HA in ocular drug delivery, many types of chemical structural modifications have been performed. In the course of our research work, we characterized nanosized cross-linked - (CLNaHA), linear sodium hyaluronate (NaHA) and zinc-hyaluronate (ZnHA), as potential ocular drug delivery systems. The aim was to determine the influence of the structure on biocompatibility, mucoadhesion and drug release. The structure was characterized by means of rheology. The cytotoxicity of the samples was determined on rabbit corneal epithelial cells (RCE) by the MTT test. Mucoadhesion measurements were made by a rheological method in vitro and by tensile tests in vitro and ex vivo. The release of sodium diclofenac, a frequently used non-steroidal anti-inflammatory drug with low bioavailability, from the gels was determined with a vertical Franz diffusion cell. The results demonstrated that all three derivatives have adequate mucoadhesive properties and their rapid drug release profiles are beneficial in ocular therapy. Thanks to these properties, the bioavailability of the ophthalmic preparations can be increased, especially with the application of CLNaHA.


Assuntos
Adesivos/química , Sistemas de Liberação de Medicamentos/métodos , Géis/química , Ácido Hialurônico/química , Nanopartículas/química , Compostos Organometálicos/química , Adesivos/efeitos adversos , Animais , Materiais Biocompatíveis/efeitos adversos , Materiais Biocompatíveis/química , Disponibilidade Biológica , Sobrevivência Celular/efeitos dos fármacos , Diclofenaco/química , Diclofenaco/farmacocinética , Sistemas de Liberação de Medicamentos/efeitos adversos , Liberação Controlada de Fármacos , Epitélio Corneano/efeitos dos fármacos , Géis/efeitos adversos , Ácido Hialurônico/efeitos adversos , Nanopartículas/efeitos adversos , Compostos Organometálicos/efeitos adversos , Coelhos , Reologia
12.
Acta Ophthalmol ; 93(6): e499-506, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25631167

RESUMO

PURPOSE: To determine the structural characteristics of lens epithelial cells (LECs) found on the anterior portion of the lens capsule and their pluripotency, proliferating and migrating potential when grown ex vivo with relevance to posterior capsular opacification (PCO) after cataract surgery. METHODS: The explants of anterior portion of the lens capsule consisting of monolayer of LECs were obtained from uneventful cataract surgery and were cultivated under adherent conditions. The size and shape of the outgrowing cells were recorded by scanning electron microscopy (SEM), while their migration and proliferation potential were followed using light microscopy. Positivity for proliferation (Ki-67)- and pluripotency (Sox2)-specific markers were tested by immunofluorescent staining. RESULTS: The proliferation and migration of anterior portion of the lens capsule's LECs filling up the denuded and reverse side regions of the lens capsule as well as their growth on glass culture surfaces could be followed by light microscopy and SEM, while the distribution of LECs and their morphology could be analysed in detail by SEM. The expression of Ki-67 and Sox2 in LECs growing adherently on human anterior portion of the lens capsule could also be detected. CONCLUSIONS: Classic light microscopy and SEM can be used to show that human anterior portion of the lens capsule harbours LECs that can proliferate and migrate, suggesting their pluripotency or putative stem cell nature. Similarly, morphological techniques can be used to study PCO and the effect different drugs or physical treatments have against PCO development.


Assuntos
Cápsula Anterior do Cristalino/ultraestrutura , Opacificação da Cápsula/prevenção & controle , Movimento Celular/fisiologia , Proliferação de Células/fisiologia , Células Epiteliais/ultraestrutura , Cristalino/ultraestrutura , Idoso , Biomarcadores/metabolismo , Extração de Catarata , Células Cultivadas , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Antígeno Ki-67/metabolismo , Microscopia , Microscopia Eletrônica de Varredura , Pessoa de Meia-Idade , Células-Tronco Pluripotentes/citologia , Fatores de Transcrição SOXB1/metabolismo
14.
Biochim Biophys Acta ; 1850(2): 435-46, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25450174

RESUMO

BACKGROUND: The apopto-phagocytic gene expression patterns during clearance of dying cells in the retina and the effect of triamcinolone (TC) upon these processes have relevance to development of age-related macular degeneration (AMD). METHODS: ARPE-19 cells and primary human retinal pigment epithelium (hRPE) were induced to undergo cell death by anoikis and the clearance of these cells by living hRPE/ARPE-19 or human monocyte-derived macrophages (HMDMs) in the presence or absence of TC was quantified by flow cytometry. TaqMan low-density gene expression array determining known markers of phagocytosis and loss-of-function studies on selected apopto-phagocytic genes was carried out in HMDM engulfing anoikic cells. RESULTS: The glucocorticoid TC had a profound phagocytosis-enhancing effect on HMDM engulfing anoikic ARPE-19 or hRPE cells, causing a selective upregulation of the Mer tyrosine kinase (MERTK) receptor, while decreasing the expression of the AXL receptor tyrosine kinase and thrombospondin-1 (THSB-1). The key role of the MERTK could be demonstrated in HMDM engulfing dying cells using gene silencing as well as blocking antibodies. Similar pathways were found upregulated in living ARPE-19 engulfing anoikic ARPE-19 cells. Gas6 treatment enhanced phagocytosis in TC-treated HMDMs. CONCLUSIONS: Specific agonists of the Mertk receptor may have a potential role as phagocytosis enhancers in the retina and serve as future targets for AMD therapy. GENERAL SIGNIFICANCE: The use of Gas6 as enhancer of retinal phagocytosis via the MerTK receptor, alone or in combination with other specific ligands of the tyrosine kinase receptors' family may have a potential role in AMD therapy.


Assuntos
Anoikis/efeitos dos fármacos , Anti-Inflamatórios/farmacologia , Células Epiteliais/enzimologia , Proteínas do Olho/biossíntese , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Macrófagos/metabolismo , Fagocitose/efeitos dos fármacos , Proteínas Proto-Oncogênicas/biossíntese , Receptores Proteína Tirosina Quinases/biossíntese , Epitélio Pigmentado da Retina/enzimologia , Triancinolona/farmacologia , Anoikis/genética , Anticorpos Neutralizantes/farmacologia , Linhagem Celular , Células Epiteliais/citologia , Proteínas do Olho/genética , Feminino , Regulação Enzimológica da Expressão Gênica/genética , Inativação Gênica/efeitos dos fármacos , Humanos , Macrófagos/citologia , Degeneração Macular/tratamento farmacológico , Degeneração Macular/enzimologia , Degeneração Macular/genética , Masculino , Fagocitose/genética , Proteínas Proto-Oncogênicas/genética , Receptores Proteína Tirosina Quinases/genética , Epitélio Pigmentado da Retina/citologia , c-Mer Tirosina Quinase
15.
J Immunol Res ; 2014: 232659, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25276840

RESUMO

A novel, simple, and reproducible method for cultivating pathological tissues obtained from human eyes during surgery was developed using viscoelastic material as a tissue adherent to facilitate cell attachment and expansion and calcium imaging of cultured cells challenged by mechanical and acetylcholine (ACh) stimulation as well as inflammatory studies. Anterior lens capsule-lens epithelial cells (aLC-LECs) from cataract surgery and proliferative diabetic retinopathy (PDR) fibrovascular epiretinal membranes (fvERMs) from human eyes were used in the study. We hereby show calcium signaling in aLC-LECs by mechanical and acetylcholine (ACh) stimulation and indicate presence of ACh receptors in these cells. Furthermore, an ex vivo study model was established for measuring the inflammatory response in fvERMs and aLC-LECs upon TNFα treatment.


Assuntos
Cálcio/metabolismo , Citocinas/metabolismo , Células Epiteliais/metabolismo , Mediadores da Inflamação/metabolismo , Técnicas de Cultura de Tecidos/métodos , Acetilcolina/farmacologia , Cápsula Anterior do Cristalino/patologia , Sinalização do Cálcio/efeitos dos fármacos , Catarata/patologia , Extração de Catarata , Adesão Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Retinopatia Diabética/patologia , Membrana Epirretiniana/patologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/patologia , Humanos , Cristalino/patologia , Microscopia de Fluorescência , Técnicas de Cultura de Tecidos/instrumentação , Substâncias Viscoelásticas/metabolismo , Substâncias Viscoelásticas/farmacologia , Vitreorretinopatia Proliferativa/patologia
16.
Biomed Res Int ; 2013: 531579, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24286080

RESUMO

Recent reports of retinal stem cells being present in several locations of the adult eye have sparked great hopes that they may be used to treat the millions of people worldwide who suffer from blindness as a result of retinal disease or injury. A population of proliferative cells derived from the ciliary body epithelium (CE) has been considered one of the prime stem cell candidates, and as such they have received much attention in recent years. However, the true nature of these cells in the adult human eye has still not been fully elucidated, and the stem cell claim has become increasingly controversial in light of new and conflicting reports. In this paper, we will try to answer the question of whether the available evidence is strong enough for the research community to conclude that the adult human CE indeed harbors stem cells.


Assuntos
Corpo Ciliar/citologia , Olho/citologia , Doenças Retinianas/terapia , Células-Tronco/citologia , Epitélio , Humanos , Retina/patologia , Retina/transplante , Doenças Retinianas/patologia , Transplante de Células-Tronco
17.
Biomed Res Int ; 2013: 492376, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24195074

RESUMO

Characterization of the cell surface marker phenotype of ex vivo cultured cells growing out of human fibrovascular epiretinal membranes (fvERMs) from proliferative diabetic retinopathy (PDR) can give insight into their function in immunity, angiogenesis, and retinal detachment. FvERMs from uneventful vitrectomies due to PDR were cultured adherently ex vivo. Surface marker analysis, release of immunity- and angiogenesis-pathway-related factors upon TNF α activation and measurement of the intracellular calcium dynamics upon mechano-stimulation using fluorescent dye Fura-2 were all performed. FvERMs formed proliferating cell monolayers when cultured ex vivo, which were negative for endothelial cell markers (CD31, VEGFR2), partially positive for hematopoietic- (CD34, CD47) and mesenchymal stem cell markers (CD73, CD90/Thy-1, and PDGFR ß ), and negative for CD105. CD146/MCAM and CD166/ALCAM, previously unreported in cells from fvERMs, were also expressed. Secretion of 11 angiogenesis-related factors (DPPIV/CD26, EG-VEGF/PK1, ET-1, IGFBP-2 and 3, IL-8/CXCL8, MCP-1/CCL2, MMP-9, PTX3/TSG-14, Serpin E1/PAI-1, Serpin F1/PEDF, TIMP-1, and TSP-1) were detected upon TNF α activation of fvERM cells. Mechano-stimulation of these cells induced intracellular calcium propagation representing functional viability and role of these cells in tractional retinal detachment, thus serving as a model for studying tractional forces present in fvERMs in PDR ex vivo.


Assuntos
Biomarcadores/análise , Retinopatia Diabética/genética , Membrana Epirretiniana/metabolismo , Proteínas de Membrana/genética , Biomarcadores/metabolismo , Técnicas de Cultura de Células , Sobrevivência Celular , Retinopatia Diabética/metabolismo , Retinopatia Diabética/patologia , Humanos , Proteínas de Membrana/classificação , Proteínas de Membrana/isolamento & purificação , Retina/metabolismo , Retina/patologia
18.
Wien Klin Wochenschr ; 124(15-16): 516-9, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22797781

RESUMO

AIM: To demonstrate a numerical data set for intraoperative pain during phacoemulsification and compare the pain scores for first and second procedures. METHODS: From 200 consecutive cases requiring bilateral cataract removals 187 were enrolled into this prospective, observational, single-surgeon, single-centre study. To evaluate the pain a 10-point visual analogue scale was used. The pain scores for both eyes of each patient were collected perioperatively (T) as well as 2-4 weeks (mean: 2.43 weeks) later, at the follow-up visit (C). Data were pooled and the four groups were compared by ANOVA All Pairweise Multiple Comparison Procedures. RESULTS: Median C-score was 1 for both eyes, T-score was 1 and 0 for the first and second eye, respectively. There wasn't any difference between the first and second eyes either in T- (1.50 ± 1.43 vs 1.51 ± 1.36) or in C-scores (0.71 vs 1.10). C-values were lower than T-values for either eye (0.71 vs 1.50 and 1.10 vs 1.51), indicating that patients recalled less pain 2-3 weeks after the surgery than that they indicated on the day of the procedure CONCLUSIONS: Consecutive phacoemulsifications do not differ in the perceived pain nevertheless, patients may believe the second eye surgery more painful because they practically compare it with the lower remembered pain for the first eye procedure. In order to avoid any disappointment we suggest warning patients before their second eye operations that they are likely to experience more pain or discomfort.


Assuntos
Dor Ocular/diagnóstico , Dor Ocular/epidemiologia , Medição da Dor/estatística & dados numéricos , Dor Pós-Operatória/diagnóstico , Dor Pós-Operatória/epidemiologia , Facoemulsificação/estatística & dados numéricos , Idoso , Feminino , Humanos , Hungria/epidemiologia , Masculino , Pessoa de Meia-Idade , Reoperação/estatística & dados numéricos , Medição de Risco , Fatores de Risco , Resultado do Tratamento
19.
Cardiovasc Ther ; 30(3): 136-44, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21106034

RESUMO

AIMS: The goal of this study was to investigate the importance of the vascular angiotensin convertase enzyme (ACE) in coronary artery bypass graft surgery (CABG) patients. METHODS: Vascular tissue (distal saphenous vein [n= 163] and/or radial artery [n= 120] segments) and blood samples were collected from CABG patients (n= 81). We studied (i) the potency of angiotensin I (AngI) and angiotensin II (AngII) to evoke vascular contractions; (ii) vascular and plasma ACE concentrations; and (iii) ACE genotype of the patients enrolled. RESULTS: The ratio of the potencies (EC(50) ) of AngII and AngI was significantly lower in radial artery compared to the saphenous vein (0.17 ± 0.03 nM and 0.51 ± 0.14 nM, respectively, P= 0.003), suggesting a 3-fold more effective AngI conversion in saphenous vein samples. Angiotensin constrictions were inhibited with telmisartan and captopril in both saphenous veins and radial arteries. Vascular ACE expression was significantly higher in saphenous vein compared to radial artery (9.7 ± 1.0 ng/mg and 5.3 ± 0.7 ng/mg, respectively, P= 0.01). Serum but no tissue ACE concentration was determined by ACE insertion/deletion polymorphism. Accordingly, no relation was found between serum and tissue ACE expression. CONCLUSION: ACE-inhibitor therapy targeting tissue located ACE may be beneficial to patients with saphenous vein grafts after CABG surgery.


Assuntos
Ponte de Artéria Coronária/efeitos adversos , Peptidil Dipeptidase A/metabolismo , Complicações Pós-Operatórias/etiologia , Veia Safena/transplante , Idoso , Bloqueadores do Receptor Tipo 1 de Angiotensina II/farmacologia , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Relação Dose-Resposta a Droga , Feminino , Humanos , Hungria , Masculino , Pessoa de Meia-Idade , Peptidil Dipeptidase A/sangue , Peptidil Dipeptidase A/genética , Complicações Pós-Operatórias/sangue , Complicações Pós-Operatórias/enzimologia , Estudos Prospectivos , Artéria Radial/efeitos dos fármacos , Artéria Radial/enzimologia , Medição de Risco , Fatores de Risco , Veia Safena/efeitos dos fármacos , Veia Safena/enzimologia , Fatores de Tempo , Resultado do Tratamento , Vasoconstritores/farmacologia
20.
Clin Exp Ophthalmol ; 39(6): 520-5, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21819505

RESUMO

BACKGROUND: To identify changing trends in indications for corneal transplantation in Debrecen, Hungary over the past 64 years. DESIGN: Retrospective study, at the Department of Ophthalmology, University of Debrecen, Hungary. PARTICIPANTS: Four thousand seven hundred and seventy-eight patients who underwent keratoplasty. METHODS: The analysis was based on medical charts, surgical descriptions and eye bank records. MAIN OUTCOME MEASURES: Keratoplasty indications between January 1946 and December 2009. For an easier overview, the 64-year interval was divided into seven time periods (1946-1955, I; 1956-1965, II; 1966-1975, III; 1976-1985, IV; 1986-1995, V; 1996-2005, VI; 2006-2009, VII). RESULTS: Over the 64 years, clinical indications for keratoplasty were corneal scarring (24.9%), regraft (18.6%), keratoconus (18.6%), pseudophakic/aphakic corneal oedema (12%), stromal corneal dystrophies (6%), non-infectious keratitis (4.7%), chemical injuries (3.3%), corneal degenerations (3%), mechanical trauma (1.7%), infectious keratitis (1.4%), endothelial corneal dystrophies (1.3%) and others (4.5%). During periods I-IV, corneal scarring was the most common indication for corneal transplantation. In period V, corneal ectasia became the leading clinical indication. Regraft was the most frequent indication in period VI. In the most recent years, an emerging tendency in pseudophakic/aphakic corneal oedema as the indication for keratoplasty was observed. CONCLUSIONS: In Hungary, the number of grafts has increased greatly in the past 64 years, the transplantation rates are similar to those of industrial countries. Indications for corneal transplantation have changed considerably over the last half decade from corneal scarring to corneal ectasia, regraft and pseudophakic/aphakic corneal oedema.


Assuntos
Doenças da Córnea/epidemiologia , Ceratoplastia Penetrante/tendências , Doenças da Córnea/cirurgia , Bancos de Olhos/estatística & dados numéricos , Hospitais Especializados/estatística & dados numéricos , Humanos , Hungria/epidemiologia , Ceratoplastia Penetrante/estatística & dados numéricos , Oftalmologia/estatística & dados numéricos , Sistema de Registros , Reoperação , Estudos Retrospectivos
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