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Posterior lamellar keratoplasty
Studený, Pavel ; Dotřelová, Dagmar (advisor) ; Pašta, Jiří (referee) ; Filipec, Martin (referee)
110 9. SUMMARY Introduction The endothelial dysfunction and related corneal oedema with decreased transparency id still the main indication for keratoplasty. Method of posterior lamellar keratoplasty, which is popular in oftalmology in last few years, is suitable method because we exchange just the affected part of the cornea. Nowadays there are more surgical ways and modalities of the technique, but the best way has not been established yet. Purpose Purpose of our study was: to evaluate and analyze the efficacy of new technique of lamella preparation, to evaluate new method of lamela implantation and to analyse anatomical outcomes in experiment and clinical outcomes in practice. Materials and methods Group 1 involves 71 eyes of 55 patients, who underwent posterior lamellar keratoplasty with implantation of a hybrid lamella DMEK-S using a hydroimplantation. In group 2 were enrolled 27 eyes of 22 patients, who underwent posterior lamellar keratoplasty with implantation of a hybrid lamella DMEK-S using glide. In the postoperative period, we evaluated corrected and uncorrected visual acuity and corneal endothelial density. We analyzed and compared clinical outcomes of both groups and we focused especially on the method of implantation. Student t test and Bonferroni test were used for all statistical analysis....
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název v anglickém jazyce není uveden
Vítová, Andrea ; Filipec, Martin (advisor) ; Rozsíval, Pavel (referee) ; Stříž, Ilja (referee)
ZÁVĚR 1) Naše výsledky rozšířily pohled na obranné mechanismy zajišťované buňkami rohovkové tkáně o schopnost stromálních buněk rohovky selektivně potlačovat produkci některých (IL-4 a IL-10) protizánětlivých cytokinů. 2) Porovnáním účinnosti léčby imunosupresivními léky nebo mAb u dvou rizikových modelů myších příjemců jsme zjistili, že pro prevenci rejekce rohovkového alotransplantátů je efektivnější léčba mAb proti CD4+ buňkám než mAb proti CD8+ buňkám či imunosupresivními léky MMF, CsA nebo jejich kombinací. Léčba pomocí mAb byla efektivní pouze u příjemců s prevaskularizací rohovky. 3) Podávání séra s obsahem cytotoxických protilátek specifických proti antigenům dárce urychlilo nástup rejekce a významně zkrátilo přežívání alotransplantátů i xenotransplantátů rohovky. Z výsledků vyplývá, že na procesu rejekce aloi xenotransplantátů rohovky se může přímo podílet protilátkami zprostředkovaná imunitní odpověď. 4) Analýzou syngenních a xenogenních transplantátů rohovky ve zvolených obdobích po transplantaci jsme zjistili, že IFN-y a NO hraje podstatnou roli při rejekci rohovkových xenotransplantátů, zatímco exprese genů pro IL-2, IL-4 a IL-10 není pro rejekci vyžadována. Tato dizertační práce se zabývala molekulárními a buněčnými mechanismy probíhajícími ve zdravé a rejekované rohovkové tkáni. Pochopení...
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Expression of endogenic lectins and their glycoligands in the tear fluid, human corneal and conjunctival epithelium under physiological and disease conditions
Hrdličková, Enkela ; Filipec, Martin (advisor) ; Heissigerová, Jarmila (referee) ; Čejková, Jitka (referee)
Purpose: Lectins play an important role in many biological processes. The aim of this work was to analyse mainly the expression of endogenic lectins, such as galectins and plant lectin, e.g. Dolichos biflorus agglutinin (DBA), and their glycoligands in the tear fluid, human corneal and conjunctival epithelium in physiological and disease conditions. Further, we studied the human natural antibody against Galα1,3Gal-R, which is mainly responsible for hyperacute rejection of xenografts transplants. We tried to investigate its localization in human corneal epithelium, lacrimal gland and tears. Material and Methods: Human tissue (lacrimal gland, tear fluid, conjunctiva, cornea, epidermis, keratinocyte and cultured corneal epithelium), as well as porcine tissue (cornea, liver and epidermis) were examined. Endogenous galectins (galectins-1, -3 and -7) were detected using immunohistochemistry methods. Binding sites for galectins, as well as binding sites for plant lectin Dolichos biflorus agglutinin, were localized by lectin histochemistry. Reverse lectin histochemistry was used for the study of binding reactivity of endogenous lectins using labelled (neo)glycoligands. Employing biotinylated natural human IgG anti -galactosides, as well as anti -galactosides, we detected reactive epitopes in human...
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Limbal stem cell transplantation and their utilization for ocular surface reconstruction.
Lenčová, Anna ; Filipec, Martin (advisor) ; Heissigerová, Jarmila (referee) ; Ardan, Taras (referee)
Aims: Limbal stem cell (LSC) deficiency is one of the most challenging ocular surface diseases. The aim of this thesis was to study damaged ocular surface reconstruction. Therefore, a mouse model of limbal transplantation was estab- lished. Furthermore, LSC isolation, transfer of LSCs and bone marrow-derived mesenchymal stem cells (MSCs) on nanofiber scaffolds were studied. Material and methods: Syngeneic, allogeneic and xenogeneic (rat) limbal grafts were transplanted orthotopically into BALB/c mice. Graft survival, immune re- sponse and the effect of monoclonal antibodies (mAb) (anti-CD4 and anti-CD8 cells) were analyzed. Mouse LSCs were separated by Percoll gradient; subse- quently, they were analyzed for the presence of LSC and differentiation corneal epithelial cell markers and characteristics using real-time PCR and flow cytom- etry. Nanofiber scaffolds seeded with LSCs and MSCs were transferred onto the damaged ocular surface in mouse and rabbit models. Cell growth on scaffolds, post-operative inflammatory response and survival of transferred cell were ana- lyzed. Results: Limbal allografts were rejected promptly by the Th1-type of immune response (IL-2, IFN-γ) involving CD4+ cells and nitric oxide produced by macro- phages, contrary to the prevailing Th1 and Th2 immune responses (IL-4, IL-10) in...
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Limbal stem cell transplantation and their utilization for ocular surface reconstruction.
Lenčová, Anna ; Filipec, Martin (advisor) ; Heissigerová, Jarmila (referee) ; Ardan, Taras (referee)
Aims: Limbal stem cell (LSC) deficiency is one of the most challenging ocular surface diseases. The aim of this thesis was to study damaged ocular surface reconstruction. Therefore, a mouse model of limbal transplantation was estab- lished. Furthermore, LSC isolation, transfer of LSCs and bone marrow-derived mesenchymal stem cells (MSCs) on nanofiber scaffolds were studied. Material and methods: Syngeneic, allogeneic and xenogeneic (rat) limbal grafts were transplanted orthotopically into BALB/c mice. Graft survival, immune re- sponse and the effect of monoclonal antibodies (mAb) (anti-CD4 and anti-CD8 cells) were analyzed. Mouse LSCs were separated by Percoll gradient; subse- quently, they were analyzed for the presence of LSC and differentiation corneal epithelial cell markers and characteristics using real-time PCR and flow cytom- etry. Nanofiber scaffolds seeded with LSCs and MSCs were transferred onto the damaged ocular surface in mouse and rabbit models. Cell growth on scaffolds, post-operative inflammatory response and survival of transferred cell were ana- lyzed. Results: Limbal allografts were rejected promptly by the Th1-type of immune response (IL-2, IFN-γ) involving CD4+ cells and nitric oxide produced by macro- phages, contrary to the prevailing Th1 and Th2 immune responses (IL-4, IL-10) in...
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Expression of endogenic lectins and their glycoligands in the tear fluid, human corneal and conjunctival epithelium under physiological and disease conditions
Hrdličková, Enkela ; Filipec, Martin (advisor) ; Heissigerová, Jarmila (referee) ; Čejková, Jitka (referee)
Purpose: Lectins play an important role in many biological processes. The aim of this work was to analyse mainly the expression of endogenic lectins, such as galectins and plant lectin, e.g. Dolichos biflorus agglutinin (DBA), and their glycoligands in the tear fluid, human corneal and conjunctival epithelium in physiological and disease conditions. Further, we studied the human natural antibody against Galα1,3Gal-R, which is mainly responsible for hyperacute rejection of xenografts transplants. We tried to investigate its localization in human corneal epithelium, lacrimal gland and tears. Material and Methods: Human tissue (lacrimal gland, tear fluid, conjunctiva, cornea, epidermis, keratinocyte and cultured corneal epithelium), as well as porcine tissue (cornea, liver and epidermis) were examined. Endogenous galectins (galectins-1, -3 and -7) were detected using immunohistochemistry methods. Binding sites for galectins, as well as binding sites for plant lectin Dolichos biflorus agglutinin, were localized by lectin histochemistry. Reverse lectin histochemistry was used for the study of binding reactivity of endogenous lectins using labelled (neo)glycoligands. Employing biotinylated natural human IgG anti -galactosides, as well as anti -galactosides, we detected reactive epitopes in human...
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Posterior lamellar keratoplasty
Studený, Pavel ; Dotřelová, Dagmar (advisor) ; Pašta, Jiří (referee) ; Filipec, Martin (referee)
110 9. SUMMARY Introduction The endothelial dysfunction and related corneal oedema with decreased transparency id still the main indication for keratoplasty. Method of posterior lamellar keratoplasty, which is popular in oftalmology in last few years, is suitable method because we exchange just the affected part of the cornea. Nowadays there are more surgical ways and modalities of the technique, but the best way has not been established yet. Purpose Purpose of our study was: to evaluate and analyze the efficacy of new technique of lamella preparation, to evaluate new method of lamela implantation and to analyse anatomical outcomes in experiment and clinical outcomes in practice. Materials and methods Group 1 involves 71 eyes of 55 patients, who underwent posterior lamellar keratoplasty with implantation of a hybrid lamella DMEK-S using a hydroimplantation. In group 2 were enrolled 27 eyes of 22 patients, who underwent posterior lamellar keratoplasty with implantation of a hybrid lamella DMEK-S using glide. In the postoperative period, we evaluated corrected and uncorrected visual acuity and corneal endothelial density. We analyzed and compared clinical outcomes of both groups and we focused especially on the method of implantation. Student t test and Bonferroni test were used for all statistical analysis....
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Genetic aspects of posterior polymorphous corneal dystrophy
Lišková, Petra ; Filipec, Martin (advisor) ; Čejková, Jitka (referee) ; Slavčev, Antonij (referee)
Posterior polymorphous corneal dystrophy (PPCD) is regarded as a rare autosomal dominant disorder, affecting both the corneal endothelium and Descemet's membrane. However, in the Czech Republic, PPCD is one of the most prevalent corneal dystrophies. The first phase of the project involved the phenotyping of Czech patients with PPCD and the collection of samples for further genetic analysis. The second phase involved determination of the PPCD chromosomal locus in two large families by using linkage analysis followed by positional candidate gene screening. In total 20 PPCD families with two or more affected members were ascertained. PPCD was diagnosed in 104 individuals, of these 82 provided peripheral blood sample for DNA isolation. Linkage analysis was performed on 52 members in two families that lead to the delineation of the PPCD locus to a 2.7 cM interval on chromosome 20p11.2, between flanking markers D20S48 and D20S139. This resulted in the exclusion of VSX1, which had previously been associated with PPCD, as the disease-causing gene in both families. Five positional candidate genes within the 2.7 cM genetic interval were screened for mutations in two probands from these families by the direct sequencing of the coding regions and no pathogenic mutations were identified. In summary, refinement of the...
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The study of the role of cell populations in rejection and tolerance of orthotopic corneal transplants
Sedláková, Klára ; Filipec, Martin (advisor) ; Jirásková, Naďa (referee) ; Čejková, Jitka (referee)
Small animal models of orthotopic corneal transplantation offer many advantages for the study of immune mechanisms after grafting - not only because of the similar mechanisms of murine and human corneal transplant rejection but also due to the feasibility of the direct observation of the animal without the need to sacrifice it. The purpose of the thesis was to study this model in mouse and rat. We established allotransplantation (BALB/c to C57BL/6 mouse and Wistar Furth to Lewis rat) and concordant xenotransplantation model (rat to mouse; Lewis to BALB/c and Sprague Dawley to BALB/c) and set up grading schemes for the evaluation of the clinical course after grafting. Initially, we focused on the effect of the suturing technique on the survival of xenografts and on the efficacy of selected immunosuppressants: cyclosporine A, monoclonal antibody against T cells (anti-Thy-1.2) and AMT (a specific inhibitor of inducible nitric oxide synthase, 2- amino-5.6-dihydro-6-methyl-4-H-1.3-thiazine)). The results demonstrate that the suturing technique significantly affects the outcome of transplantation and, importantly, influences the effectiveness of immunosuppressive regimens and therefore must be taken into account when evaluating the efficacy of immunosuppressive drugs. FTY720 is a novel immunosuppressant with a...
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