Mesenchymal stem cells (MSCs) represent a population of adult stem cells that have potent immunoregulatory, anti-inflammatory, and antiapoptotic properties. In addition, they have ability to migrate to the site of inflammation or injury, where they contribute to the regeneration and healing process. For these properties, MSCs have been used as therapeutic cells in several models, including treatment of damages or disorders of the ocular surface. If the damage of the ocular surface is extensive and involves a limbal region where limbal stem cell reside, MSC therapy has been proved as the effective treatment approach. Although the anti-inflammatory properties of MSCs have been well characterized, mechanisms of antiapoptotic action of MSCs are not well recognized. Using a chemically damaged cornea in a mouse model, we showed that the injury decreases expression of the gene for antiapoptotic molecule Bcl-2 and increases the expression of proapoptotic genes Bax and p53. These changes were attenuated by local transplantation of MSCs after corneal damage. The antiapoptotic effect of MSCs was tested in an in vitro model of co-cultivation of corneal explants with MSCs. The apoptosis of corneal cells in the explants was induced by proinflammatory cytokines and was significantly inhibited in the presence of MSCs. The antiapoptotic effect of MSCs was mediated by paracrine action, as confirmed by separation of the explants in inserts or by supernatants from MSCs. In addition, MSCs decreased the expression of genes for the molecules associated with endoplasmic reticulum stress Atf4, Bip, and p21, which are associated with apoptosis. The results show that MSCs inhibit the expression of proapoptotic genes and decrease the number of apoptotic cells in the damaged corneas, and this action might be one of the mechanisms of the therapeutic action of MSCs.

• Klíčová slova
• Bax, Bcl-2, antiapoptotic properties, cornea, mesenchymal stem cells, mouse model,
• MeSH
• apoptóza genetika   MeSH
• cyklooxygenasa 2 genetika   metabolismus   MeSH
• cytokiny genetika   metabolismus   MeSH
• hepatocytární růstový faktor genetika   metabolismus   MeSH
• keratitida genetika   metabolismus   patologie   MeSH
• kultivované buňky MeSH
• lidé MeSH
• mezenchymální kmenové buňky cytologie   metabolismus   MeSH
• modely nemocí na zvířatech * MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• poranění rohovky genetika   metabolismus   terapie   MeSH
• regulace genové exprese * MeSH
• rohovka metabolismus   MeSH
• transplantace mezenchymálních kmenových buněk metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cyklooxygenasa 2 MeSH
• cytokiny MeSH
• hepatocytární růstový faktor MeSH

The aim of this study is to examine whether molecular hydrogen (H2) is able to reduce oxidative stress after corneal damage induced by UVB irradiation. We previously found that UVB irradiation of the cornea caused the imbalance between the antioxidant and prooxidant enzymes in the corneal epithelium, followed by the imbalance between metalloproteinases and their physiological inhibitors (imbalances in favour of prooxidants and metalloproteinases) contributing to oxidative stress and development of the intracorneal inflammation. Here we investigate the effect of H2 dissolved in PBS in the concentration 0.5 ppm wt/vol, applied on rabbit corneas during UVB irradiation and healing (UVB doses 1.01 J/cm2 once daily for four days). Some irradiated corneas remained untreated or buffer treated. In these corneas the oxidative stress appeared, followed by the excessive inflammation. Malondiladehyde and peroxynitrite expressions were present. The corneas healed with scar formation and neovascularization. In contrast, in H2 treated irradiated corneas oxidative stress was suppressed and malondiladehyde and peroxynitrite expressions were absent. The corneas healed with the restoration of transparency. The study provides the first evidence of the role of H2 in prevention of oxidative and nitrosative stress in UVB irradiated corneas, which may represent a novel prophylactic approach to corneal photodamage.

• MeSH
• králíci MeSH
• kyselina peroxydusitá metabolismus   MeSH
• malondialdehyd metabolismus   MeSH
• oxidační stres účinky léků   účinky záření   MeSH
• poranění rohovky farmakoterapie   metabolismus   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• rohovka účinky léků   metabolismus   účinky záření   MeSH
• ultrafialové záření * MeSH
• vodík terapeutické užití   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• publikace stažené z tisku MeSH
• Názvy látek
• kyselina peroxydusitá MeSH
• malondialdehyd MeSH
• reaktivní formy kyslíku MeSH
• vodík MeSH

The aim of this study was to examine the effect of molecular hydrogen (H2) on the healing of alkali-injured cornea. The effects of the solution of H2 in phosphate buffered saline (PBS) or PBS alone topically applied on the alkali-injured rabbit cornea with 0.25 M NaOH were investigated using immunohistochemical and biochemical methods. Central corneal thickness taken as an index of corneal hydration was measured with an ultrasonic pachymeter. Results show that irrigation of the damaged eyes with H2 solution immediately after the injury and then within next five days renewed corneal transparency lost after the injury and reduced corneal hydration increased after the injury to physiological levels within ten days after the injury. In contrast, in injured corneas treated with PBS, the transparency of damaged corneas remained lost and corneal hydration elevated. Later results-on day 20 after the injury-showed that in alkali-injured corneas treated with H2 solution the expression of proinflammatory cytokines, peroxynitrite, detected by nitrotyrosine residues (NT), and malondialdehyde (MDA) expressions were very low or absent compared to PBS treated injured corneas, where NT and MDA expressions were present. In conclusion, H2 solution favorably influenced corneal healing after alkali injury via suppression of oxidative stress.

• MeSH
• aktiny metabolismus   MeSH
• cytokiny metabolismus   MeSH
• exprese genu účinky léků   MeSH
• hydroxid sodný toxicita   MeSH
• imunohistochemie MeSH
• interleukin-1beta genetika   metabolismus   MeSH
• keratin-12 metabolismus   MeSH
• keratin-3 metabolismus   MeSH
• králíci MeSH
• kyselina peroxydusitá metabolismus   MeSH
• malondialdehyd metabolismus   MeSH
• modely nemocí na zvířatech MeSH
• oxidační stres účinky léků   MeSH
• poranění rohovky etiologie   metabolismus   patologie   MeSH
• rohovka metabolismus   patologie   MeSH
• vaskulární endoteliální růstový faktor A genetika   metabolismus   MeSH
• vodík farmakologie   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• publikace stažené z tisku MeSH
• Názvy látek
• aktiny MeSH
• cytokiny MeSH
• hydroxid sodný MeSH
• interleukin-1beta MeSH
• keratin-12 MeSH
• keratin-3 MeSH
• kyselina peroxydusitá MeSH
• malondialdehyd MeSH
• vaskulární endoteliální růstový faktor A MeSH
• vodík MeSH

In this study we tried to develop a new approach to suppress inflammation and neovascularization in the alkali-injured rabbit cornea. For this reason Cyclosporine A (CsA)-loaded electrospun nanofibers were transferred onto the ocular surface injured with alkali (0.25 N NaOH). Damaged corneas were divided into the following groups: untreated, treated with CsA eye drops, treated with nanofibers drug-free and treated with CsA-loaded nanofibers. Healthy rabbit corneas served as controls. Drug-free nanofibers and CsA-loaded nanofibers were transferred onto the damaged corneal surface immediately after the injury and sutured to conjunctiva. On day five after the injury the nanofibers were removed. The animals from all groups were sacrificed on day twelve after the injury. The extent of the inflammatory reaction and corneal healing were examined macroscopically, immunohistochemically and biochemically. The central corneal thickness was measured using an ultrasonic pachymeter. When compared with untreated injured corneas, injured corneas treated with drug-free nanofibers or injured corneas treated with CsA eye drops, the number of CD3-positive cells (T lymphocytes) and the production of pro-inflammatory cytokines were strongly reduced in corneas treated with CsA-loaded nanofibers, which was associated with the significantly decreased expression of matrix metalloproteinase 9, inducible nitric oxide synthase, vascular endothelial growth factor and active caspase-3. CsA-loaded nanofibers effectively suppressed corneal inflammation and corneal neovascularization. Central corneal thickness restored to levels before injury only in corneas treated with CsA-loaded nanofibers. Corneal transparency was highly restored in these corneas. It is suggested that the beneficial effect of CsA-loaded nanofibers was associated with the continuous release of CsA from nanofibers and continuous affection of damaged cornea by CsA. The suture of nanofibers to conjunctiva and the closed eyes contributed to beneficial corneal healing. This is in contrast to CsA eye drops, which are quickly washed from the ocular surface and the contact of CsA with the damaged cornea was limited. In conclusion, the approach with CsA-loaded nanofibers could represent an effective alternative mode of therapy for corneal chemical burns.

• Klíčová slova
• Alkali injury, Central corneal thickness, CsA-loaded nanofibers, Immunohistochemistry, Inflammation, Rabbit cornea, Real time PCR,
• MeSH
• alkálie škodlivé účinky   MeSH
• chemické popálení farmakoterapie   MeSH
• cyklosporin aplikace a dávkování   terapeutické užití   MeSH
• imunohistochemie MeSH
• imunosupresiva terapeutické užití   MeSH
• kaspasa 3 metabolismus   MeSH
• králíci MeSH
• modely nemocí na zvířatech MeSH
• nanovlákna MeSH
• neovaskularizace rohovky patologie   MeSH
• popálení oka farmakoterapie   metabolismus   patologie   MeSH
• poranění rohovky chemicky indukované   metabolismus   patologie   MeSH
• T-lymfocyty patologie   MeSH
• vaskulární endoteliální růstový faktor A metabolismus   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• alkálie MeSH
• cyklosporin MeSH
• imunosupresiva MeSH
• kaspasa 3 MeSH
• vaskulární endoteliální růstový faktor A 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
• Názvy látek
• malondialdehyd MeSH

Oxidative stress is involved in many ocular diseases and injuries. The imbalance between oxidants and antioxidants in favour of oxidants (oxidative stress) leads to the damage and may be highly involved in ocular aging processes. The anterior eye segment and mainly the cornea are directly exposed to noxae of external environment, such as air pollution, radiation, cigarette smoke, vapors or gases from household cleaning products, chemical burns from splashes of industrial chemicals, and danger from potential oxidative damage evoked by them. Oxidative stress may initiate or develop ocular injury resulting in decreased visual acuity or even vision loss. The role of oxidative stress in the pathogenesis of ocular diseases with particular attention to oxidative stress in the cornea and changes in corneal optical properties are discussed. Advances in the treatment of corneal oxidative injuries or diseases are shown.

• MeSH
• alkálie toxicita   MeSH
• oční roztoky terapeutické užití   MeSH
• oxidační stres * účinky léků   účinky záření   MeSH
• poranění rohovky farmakoterapie   metabolismus   patologie   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• rohovka účinky léků   účinky záření   MeSH
• ultrafialové záření MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• alkálie MeSH
• oční roztoky MeSH
• reaktivní formy kyslíku MeSH