V patologii očního povrchu při suchém oku mohou hrát důležitou úlohu oxyradikály a destruktivní proteázy. Aktivní urokináza se v slzách normálního oka nevyskytuje, aproto nález aktivní formy tohoto enzymu je ukazatelem iniciace patologických změn. Nové metody otiskové cytologie užívající enzymy jako márkry metabolických poruch zlepší diagnostiku suchého oka. Biochemické metody, užívající citlivé fluorescenční substráty, jsou vhodné pro vyšetření přítomnosti i koncentrace destruktivních proteáz v slznémfilmu.; The dry eye syndrome may be associated with increased amount of oxyradicals and destructive proteases at the ocular surface. Active urokinase is not present in the tear fluid of the normal human eye. The finding of active urokinase may serve as marker ofocular disorders. New methods of impression cytology using enzymes as markers of metabolic disturbances can improve diagnostic purposes in humans with dry eye. Biochemical methods using specific fluorogenic substrates enable us to detect the appearanceand concentration of protease activities in tears.

• MeSH
• aktivátor plazminogenu urokinázového typu analýza   MeSH
• antioxidancia MeSH
• biologické markery MeSH
• biologie buňky MeSH
• syndromy suchého oka diagnóza   terapie   MeSH

• Konspekt
• Ortopedie. Chirurgie. Oftalmologie
• NLK Obory
• oftalmologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

Vliv oxidativního a hypoxického stresu na stárnutí oka a způsoby prevence předčasného stárnutí pomocí nových antioxidačních agens a kontaktních čoček minimalizujících hypoxický a mechanický ster na oko. Věkové rozdíly v hojení očních lézí.Enzymy jako márkry metabolických změn, změny jejich spektra či aktivit v závislosti na stáří.Jejich detekce in situ (oční tkáně) a extra situm (slzy,tekutina přední oční komory).; The rle of oxidative,hypoxic and mechanical stress on aging of the eye.Pramature aging of the eye and new approaches of its prevention by means of antioxidant agents and newly developed contact lenses producing minimal hypoxic and mechanical stress. Age-related differences in the healing of eye lesions,enzymes as markers of metabolic eye disturbances,age-related differences in enzyme pattern.Their "in situ" (anterior eye segment) and "extra situm" (tears,aqueous humor) detection.

• MeSH
• kontaktní čočky škodlivé účinky   MeSH
• mechanický stres MeSH
• oční symptomy MeSH
• oxidační stres MeSH
• stárnutí MeSH

• Konspekt
• Ortopedie. Chirurgie. Oftalmologie
• NLK Obory
• oftalmologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

Diagnostika příčin špatného hojení ran povrchu oka a kůže pomocí enzymatických márkrů a usnadnění hojení užitím hydrofilních polymerních materiálů jako provizorních krytů a nosičů lékových substancí,zejména růstových faktorů a inhibitorů proteáz.

• MeSH
• antioxidancia MeSH
• beta-galaktosidasa antagonisté a inhibitory   MeSH
• hojení ran MeSH
• kathepsin B antagonisté a inhibitory   MeSH
• kontaktní čočky MeSH
• lékové transportní systémy MeSH
• volné radikály antagonisté a inhibitory   MeSH

• Konspekt
• Lékařské vědy. Lékařství
• NLK Obory
• oftalmologie
• dermatovenerologie
• dermatovenerologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

The aim of this study was to examine whether nanofiber scaffolds seeded with rabbit bone marrow mesenchymal stem cells (MSCs nanofibers) transferred onto the damaged corneal surface and covered with cyclosporine A (CsA)-loaded nanofiber scaffolds (CsA nanofibers) enable healing of the rabbit cornea injured with 1N NaOH. The healing of damaged corneas was examined morphologically, immunohistochemically and biochemically on day 24 after the injury. Compared to untreated injured corneas, where corneal ulceration or large corneal thinning or even perforation were developed, injured corneas treated with drug free nanofibers healed without profound disturbances in a majority of cases, although with fibrosis and scar formation. In injured corneas treated with CsA nanofibers or MSCs nanofibers, the development of scar formation was reduced. Best healing results were obtained with a combination of MSCs and CsA nanofibers (MSCs-CsA nanofibers). Corneas healed with highly restored transparency. Neovascularization highly expressed in untreated injured corneas and reduced in corneas treated with CsA nanofibers or MSCs nanofibers, was suppressed in corneas treated with MSCs-CsA nanofibers. The levels of matrix metalloproteinase 9, inducible nitric oxide synthase, interleukin 6, α-smooth muscle actin, tumor growth factor β and vascular endothelial growth factor were significantly decreased in these corneas as compared to untreated corneas, where the levels of the above mentioned markers were high. In conclusion, MSCs-CsA nanofibers were effective in the treatment of severe alkali-induced corneal injury.

• MeSH
• cyklosporin aplikace a dávkování   MeSH
• hojení ran účinky léků   MeSH
• hydroxid sodný toxicita   MeSH
• imunosupresiva aplikace a dávkování   MeSH
• jizva prevence a kontrola   MeSH
• kaustika toxicita   MeSH
• králíci MeSH
• lékové transportní systémy metody   MeSH
• modely nemocí na zvířatech MeSH
• nanovlákna MeSH
• neovaskularizace rohovky prevence a kontrola   MeSH
• nosiče léků MeSH
• poranění rohovky terapie   MeSH
• tkáňové podpůrné struktury MeSH
• transplantace mezenchymálních kmenových buněk metody   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

UNLABELLED: Stem cell-based therapy has become an attractive and promising approach for the treatment of severe injuries or thus-far incurable diseases. However, the use of stem cells is often limited by a shortage of available tissue-specific stem cells; therefore, other sources of stem cells are being investigated and tested. In this respect, mesenchymal stromal/stem cells (MSCs) have proven to be a promising stem cell type. In the present study, we prepared MSCs from bone marrow (BM-MSCs) or adipose tissue (Ad-MSCs) as well as limbal epithelial stem cells (LSCs), and their growth, differentiation, and secretory properties were compared. The cells were grown on nanofiber scaffolds and transferred onto the alkali-injured eye in a rabbit model, and their therapeutic potential was characterized. We found that BM-MSCs and tissue-specific LSCs had similar therapeutic effects. Clinical characterization of the healing process, as well as the evaluation of corneal thickness, re-epithelialization, neovascularization, and the suppression of a local inflammatory reaction, were comparable in the BM-MSC- and LSC-treated eyes, but results were significantly better than in injured, untreated eyes or in eyes treated with a nanofiber scaffold alone or with a nanofiber scaffold seeded with Ad-MSCs. Taken together, the results show that BM-MSCs' therapeutic effect on healing of injured corneal surface is comparable to that of tissue-specific LSCs. We suggest that BM-MSCs can be used for ocular surface regeneration in cases when autologous LSCs are absent or difficult to obtain. SIGNIFICANCE: Damage of ocular surface represents one of the most common causes of impaired vision or even blindness. Cell therapy, based on transplantation of stem cells, is an optimal treatment. However, if limbal stem cells (LSCs) are not available, other sources of stem cells are tested. Mesenchymal stem cells (MSCs) are a convenient type of cell for stem cell therapy. The therapeutic potential of LSCs and MSCs was compared in an experimental model of corneal injury, and healing was observed following chemical injury. MSCs and tissue-specific LSCs had similar therapeutic effects. The results suggest that bone marrow-derived MSCs can be used for ocular surface regeneration in cases when autologous LSCs are absent or difficult to obtain.

• MeSH
• biologické markery metabolismus   MeSH
• buněčná a tkáňová terapie metody   MeSH
• buněčná diferenciace MeSH
• buňky kostní dřeně cytologie   fyziologie   MeSH
• chemické popálení patologie   terapie   MeSH
• epitelové buňky cytologie   fyziologie   transplantace   MeSH
• exprese genu MeSH
• fyziologická neovaskularizace MeSH
• králíci MeSH
• limbus corneae krevní zásobení   zranění   MeSH
• mezenchymální kmenové buňky cytologie   fyziologie   MeSH
• primární buněčná kultura MeSH
• proliferace buněk MeSH
• reepitalizace fyziologie   MeSH
• rohovkový epitel krevní zásobení   zranění   MeSH
• tkáňové podpůrné struktury MeSH
• transplantace mezenchymálních kmenových buněk * MeSH
• tuková tkáň cytologie   fyziologie   MeSH
• tukové buňky cytologie   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH

In various corneal injuries (such as chemical burns or irradiation of corneas with UVB radiation) and ocular diseases (e.g. dry eye disease, keratokonus, bullous keratopathy, Fuchs' endothelial dystrophy), the expressions of malondialdehyde (a marker of lipid peroxidation) and nitrotyrosine (a marker of oxidative stress) appeared in cells of individual corneal layers and conjunctival cells (dry eye disease). This is in contrast to healthy corneas in which negligible levels of malondialdehyde and no expressions of nitrotyrosine are present. The injured or diseased corneas reveal decreased capacity of antioxidants (enzymatic as well as non-enzymatic), whereas the levels of pro-oxidants (e.g. oxidases that generate reactive oxygen species) remain at physiological levels or even increase, leading to the antioxidant/prooxidant imbalance and oxidative stress. Oxidative stress in the cornea stimulates generation of pro-inflammatory cytokines, proteolytic enzymes and enzymes that generate nitric oxide (nitric oxide synthases). An abundant amout of reactive oxygen species and nitric oxide lead to the formation of toxic reactive products contributing to tissue damage. This review aims to summarize immunohistochemical changes in severe corneal injuries and diseases in which oxidative stress has been proved.

• MeSH
• lidé MeSH
• malondialdehyd metabolismus   MeSH
• nemoci rohovky metabolismus   patologie   MeSH
• oxidační stres * MeSH
• poranění rohovky metabolismus   patologie   MeSH
• radiační poranění patologie   MeSH
• rohovka metabolismus   patologie   MeSH
• ultrafialové záření MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

UVB radiation from sunlight induces an acute corneal inflammation, photokeratitis, accompanied by changes in corneal hydration. We employed a method of ultrasonic pachymetry for daily examination of central corneal thickness as an index of corneal hydration of the rabbit cornea repeatedly irradiated by UVB radiation (312 nm, daily dose of 0.25 J/cm(2) during three or four days) as influenced by UVB absorber (actinoquinol combined with hyaluronic acid) dropped on the ocular surface during irradiation. One day after the third irradiation procedure the animals were sacrificed and corneas examined immuno-histochemically for peroxynitrite formation, a marker of oxidative damage, the antioxidant aldehyde dehydrogenase 3A1 and endothelial nitric oxide synthase, an enzyme generated nitric oxide. Results show that UV absorber combined with hyaluronic acid protected the cornea against UVB-induced changes in corneal thickness and microscopical disturbances to the cornea (both seen after buffered saline application) until the fourth experimental day. These UVB doses are equivalent to a daily exposure of 2.5 hrs of the human cornea to solar UVB radiation for three consecutive days. It is suggested that actinoquinol/ hyaluronic acid drops might be helpful for the human eye in the defence against photooxidative and other oxidative processes.

• MeSH
• aldehyddehydrogenasa metabolismus   MeSH
• aplikace oční MeSH
• časové faktory MeSH
• chinoliny aplikace a dávkování   chemie   MeSH
• cytoprotekce MeSH
• edém rohovky metabolismus   prevence a kontrola   ultrasonografie   MeSH
• experimentální radiační poranění metabolismus   prevence a kontrola   ultrasonografie   MeSH
• imunohistochemie MeSH
• keratitida metabolismus   prevence a kontrola   ultrasonografie   MeSH
• králíci MeSH
• kyselina hyaluronová aplikace a dávkování   chemie   MeSH
• kyselina peroxydusitá metabolismus   MeSH
• radioprotektivní látky aplikace a dávkování   chemie   MeSH
• rohovka metabolismus   účinky léků   ultrasonografie   MeSH
• spektrofotometrie ultrafialová MeSH
• synthasa oxidu dusnatého, typ III metabolismus   MeSH
• tělesná voda metabolismus   MeSH
• ultrafialové záření MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: Trehalose, a nonreducing disaccharide of glucose, is synthesized as a stress response factor when cells are exposed to stressful conditions. In the cornea, oxidative stress plays the key role in the development of acute corneal inflammatory response to UVB rays, photokeratitis. We found previously that trehalose reduced UVB-induced oxidative effects on the formation of cytotoxic peroxynitrite, apoptotic corneal epithelial cell death and changes in corneal optics. The aim of the present study was to examine whether trehalose might inhibit UVB-mediated proinflammatory cytokine and matrix metalloproteinase induction and the development of an antioxidant/pro-oxidant imbalance in the corneal epithelium, changes found previously to be strongly involved in the acute corneal UVB-induced inflammation. The expression of heat shock protein 70 as a potential biomarker for corneal UVB-induced damage was also examined. METHODS: The corneas of New Zealand white rabbits were irradiated with UVB rays, 312 nm, daily dose of 0.5 J/cm(2) for 4 days. During the irradiation, trehalose drops were applied on the right eye and buffered saline on the left eye. One day after the end of irradiations, the animals were killed and the corneas examined immunohistochemically for the expression of antioxidant enzymes (catalase, superoxide dismutase, glutathione peroxidase), pro-oxidant xanthine oxidoreductase/xanthine oxidase, proinflammatory cytokines (interleukin-6, interleukin-8), matrix metalloproteinase-9 and heat shock protein 70. RESULTS: After buffered saline treatment during UVB irradiation, an antioxidant/pro-oxidant imbalance appeared in the corneal epithelium: The expression of antioxidant enzymes was highly reduced, whereas the expression of pro-oxidant xanthine oxidase was increased. The pronounced expression of pro-inflammatory cytokines, matrix metalloproteinase and heat shock protein 70 was found in the UVB-irradiated corneal epithelium. Trehalose application significantly suppressed all the above-mentioned UVB-induced corneal disturbances. CONCLUSIONS: Trehalose favorably influenced the oxidative damage of the cornea caused by UVB rays. Trehalose suppressed proinflammatory cytokine induction. It is suggested that suppression of proinflammatory cytokines contributed strongly to reduced matrix metalloproteinase and xanthine oxidase expression in the UVB-irradiated corneal epithelium and to the decreased development of an antioxidant/pro-oxidant imbalance. The overexpression of heat shock protein 70 found in UVB-irradiated cornea after buffered saline treatment was reduced after trehalose application.

• MeSH
• antioxidancia MeSH
• biologické markery metabolismus   MeSH
• cytokiny metabolismus   MeSH
• experimentální radiační poranění farmakoterapie   enzymologie   MeSH
• imunoenzymatické techniky MeSH
• králíci MeSH
• matrixová metaloproteinasa 9 metabolismus   MeSH
• oxidační stres účinky léků   MeSH
• oxidancia MeSH
• oxidoreduktasy metabolismus   MeSH
• počítačové zpracování obrazu MeSH
• proteiny tepelného šoku HSP70 metabolismus   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• rohovkový epitel enzymologie   účinky záření   MeSH
• trehalosa farmakologie   MeSH
• ultrafialové záření škodlivé účinky   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

PURPOSE: Normal corneal hydration is necessary for the maintenance of corneal transparency. Damage of the corneal epithelium or endothelium by various external influences disturbs the mechanism by which the cornea maintains normal hydration and transparency. The cornea swells, and the corneal thickness increases, resulting in increased scatter and the development of corneal opacity. The transmission of light across the cornea is changed. The purpose of this study is to investigate spectrophotometrically the corneal light transmission under the influence of the various factors affecting the cornea. METHODS: We developed a spectrophotometric method to measure the light transmission across the cornea under the influence of various factors affecting the cornea, such as treatment with 0.9% NaCl, saline, or phosphate buffered saline (PBS), solutions employed as placebo eye drops (negative controls) in experimental studies, agents toxic to the cornea, such as diluted acids or alkalis. The method distinguishes between changes in corneal light transmission caused by altered corneal thickness (the level of hydration) and changes resulting from other corneal disturbances which in turn affect corneal light transmission. RESULTS: The results obtained show that the corneal light transmission is decreased following the application of toxic substances on the corneal surface. This decrease is highly dependent on the severity of the corneal injury evoked by individual noxes, and the resulting changes in corneal hydration and transparency. CONCLUSIONS: The influence of various influences applied to the cornea, manifested as changes in corneal light transmission, can be measured using our spectrophotometric method with a high degree of sensitivity.

• MeSH
• edém rohovky chemicky indukované   MeSH
• hydroxid sodný toxicita   MeSH
• králíci MeSH
• kyselina chlorovodíková toxicita   MeSH
• rohovka účinky léků   účinky záření   MeSH
• spektrofotometrie MeSH
• světlo MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The aim of this study was to compare the presence and activity of matrix metalloproteinases (MMPs) 1, 2, 3, 7, 8, 9 and 13 in human melted and cadaverous corneas. Twelve melted corneal specimens from three patients with rheumatoid arthritis, one patient with ocular cicatricial pemphigoid and one patient with melting attributed to spastic entropion and ten control corneal buttons were used. The presence of MMPs was detected using indirect enzyme immunohistochemistry. The active forms of MMP-2 and -9 and MMP-3 and -7 were examined by gelatin and casein zymography, respectively. The concentrations of active MMP-1 and -3 were measured using activity assays. Increased immunostaining intensity for MMP-1 and -9 was seen in the corneal epithelium and the anterior stroma of all, and for MMP-2, -3, -7 and -8 of almost all, melted corneas compared to the negative or slightly positive staining of the controls. The posterior stroma showed the presence of MMP-1, -2, -3 and -9 in almost all and of MMP-7 and -8 in half of all melted specimens. A markedly higher level of active MMP-2 was detected in six and active MMP-9 in all of eleven pathologic specimens compared to control specimens, using gelatin zymography. The proenzymes of MMP-3 and -7 and the MMP-7 intermediate cleavage product were detected only in melted corneas using casein zymography. Significantly increased MMP-1 and -3 activity was also found in the melted corneas using activity assays. The markedly increased immunostaining for MMP-1, -2, -3, -7, -8 and -9 as well as the elevated levels of the active forms of MMP-1, -2, -3 and -9 in melted corneal specimens from patients with various diagnoses suggest that although different stimuli may trigger the pathways that lead to the destruction of the extracellular matrix, these enzymes could play a subsequent role in this process.

• MeSH
• benigní pemfigoid sliznice komplikace   MeSH
• dárci tkání MeSH
• dospělí MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• imunoenzymatické techniky MeSH
• keratoplastika perforující MeSH
• lidé středního věku MeSH
• lidé MeSH
• matrixové metaloproteinasy fyziologie   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• nemoci rohovky enzymologie   chirurgie   MeSH
• recidiva MeSH
• revmatoidní artritida komplikace   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH