Poster Sessions Saturday

Introduction: According to authors, treatment of pterygium is limited to its simple excision. Despite the meticulous technique of different operations, none of them excludes post-operative complications, such as imperfect adaptation of conjunctiva on the previous pterygium place, development of post-op astigmatism, pterygium recurrence. Pterygium recurrence usually happens during first year after the surgery, more often first 6 months. The recurrence appears in more than 40-70% by different authors. Purpose: To study the effectiveness of pterygium surgical treatment with high-frequency electrowelding of biologic tissues for the fixation of free limbal-conjunctival autograft (LCAG). Material and methods: Surgical treatment data of 105 patients with pterygium were analyzed in the study. 73 patients (73 eyes) were a research group, which underwent a free LCAG according to developed surgical procedure (patent of Ukraine). 32 patients (32 eyes) made a group of comparison; they were operated by Mc-Reynolds, Arlt etc. Patients from the research group were divided to the basic and control groups. 39 patients (39 eyes) from control group - was performed using with suture fixing free LCAG. 34 patients (34 eyes) from basic group underwent of high-frequency electrowelding of biologic tissues for the fixation of free LCAG using original designed bipolar forceps of the modified device ЕК-300М1. Results: Clinical manifestations of corneal syndrome in the early postoperative period were observed in 9 patients on 9 eyes (26,5%) of basic group, compared to control group, where corneal syndrome occurred in 33 patients on 33 eyes (84,6%). A complete corneal epithelization was registered in 30 patients on 30 eyes (88,2%) of the basic group, compared to control group in 20 patients (51,3%). At observation terms of 6 months a recurrent pterygium was found in 6 patients (8,2%) of the group with free LCAG. In the control group a recurrent pterygium occurred on 9 eyes (28,1%). Conclusion: Based on our study investigated the possibility of surgical treatment in pterihium using free LCAG, which is statistically significantly reduces the risk of recurrence of the disease 3.4 times (95% CI 1,1 ÷ 10,4) compared to other methods. The use of high-frequency electric tissue fixation for free LCAG reduces the incidence of clinical manifestations of corneal syndrome in 58,1% (p < 0,001), increase the frequency of complete epithelialization of the cornea to 36,9% (p = 0,0017).

Purpose: To evaluate and investigate the effect of subconjuctival injection of anti-VEGF (Bevacizumab) to prevent recurrence of pterygium. Method: Twenty patients (mean age : 33 +/- 4 year old, women were 25 % and Egyptian ancestry was 100% of the cases) with 20 eyes with primary pterygium were included in this work. Exclusion criteria included recurrent pterygium or any other corneal and/or conjuctival diseases. Patients underwent pterygium excision with local anesthesia and intraoperative subconjuctival 0.1 mL (1 mg) anti-VEGF (Bevacizumab). The main outcome investigated was the recurrence of pterygium and corneal vascularization at any time during postoperative follow-up. Results: The 20 eyes of the 20 patients were followed up for an average of 6 +/- 3 months. Recurrence of pterygium and corneal vascularization in 1 eye (5 %). Conclusion: In this work, it was clear that intraoperative subconjunctival anti-VEGF (Bevacizumab) has influenced the frequency of non-recurrence of pterygium and corneal vascularization after primary pterygium surgical excision. Further works and investigations will also be needed.

Purpose: Dry-eye disease (DED) is one of the most common pathological conditions of the eye, affecting millions of patients. In particular evaporation and meibomian gland dysfunction are discussed as core reasons for dry eye disease (DED. Recently the novel substance perfluorohexyloctane (F6H8), a semifluorinated alkane (SFA), has been introduced as a non-blurring, preservative-free wetting agent that reduces shearing forces between surfaces and enables to bind lipids due to its lipophilic nature without the necessity to form water-oil suspensions. After successful pre-clinical testing F6H8 has been approved as a medical device (NovaTears®) and was used in an observational study in patients with mild to moderate DED. Methods: 30 Patients with evaporative DED were included in an open, prospective, uncontrolled post-market clinical follow-up study and, after providing their informed consent, applied 4 times daily in both eyes for 6 weeks. Clinical examination included visual acuity, intraocular pressure (IOP), Schirmer I, tearfilm break-up time (TFBUT), corneal fluorescein staining and meibum secretion, etc. In addition each patient completed a questionnaire similar to the Ocular surface disease index (OSDI©). Results: In this trial, visual acuity and IOP did not change over time, Schirmer I increased significantly from 10.3 (+/- 3.9) to 16.2 (+/- 9.1) P =0.0011 (mean of both eyes). TFBUT increased significantly from 6.0 (+/- 2.5) to 8.8 (+/- 4.9) P = 0.0026 in the right eye and from 5.8 (+/- 2.6) to 9.6 (+/- 5.9) P = 0.0006 in the left eye, respectively. Ocular surface staining decreased significantly (P = 0.0013 in the right eye and P = 0.0041 in the left eye and OSDI decreased from 55 (+/- 23) to 34 (+/- 22) P < 0.0001. Symptoms such as redness, itching, etc. improved whereas meibum quality only improved in some patients. Conclusions: This first prospective treatment survey shows that patients suffering from hyperevaporative DED benefit from using NovaTears®. In particular, tearfilm stability and tear production increased, while corneal staining and OSDI score decreased significantly over the time course of the trial. NovaTears® was well tolerated with only low numbers of adverse events. Overall this trial demonstrates a safe application of this new type of unpreserved artificial tear in mild to moderate evaporative DED.

Fragestellung: Vergleichende Bewertung des Einflusses von Tränenersatzmitteln auf den kornealen Heilungsprozess und deren korneale Toxizität. Methodik: Optive® und Optive Fusion® wurden bezüglich ihrer kornealen Reizung gegen Vismed Multi® als Positivkontrolle und Benzalkoniumchlorid 0,01 % (BAK) als Negativkontrolle getestet. Die Formulierungen (0.72-1.2 ml pro Tag) wurden auf Kaninchenhornhäuten (n=5) auf künstlichen Vorderkammern (EVEIT) kultiviert und stündlich über 6 Tage appliziert. Initial wurden 4 korneale Erosiones (2.4 – 4.6 mm²) induziert. Die Erosiones wurden während der Applikation mittels Fluoreszeinfärbungen und Fotos überwacht. Um die Vitalität der Corneae zu bestimmen, wurden die Glukose und Laktatlactatkonzentrationen in der Vorderkammerflüssigkeit bestimmt. Die korneale Fluoreszeinpermeabilität wurde getestet als Indikator für die korneale Barrierefunktion. Ergebnis: Alle Produkte zeigten eine komplette korneale Heilung an Tag 2. Im Anschluss entwickelten alle 5 Optive®-behandelten Corneae eine progressive, fluoreszein-positive Erosion (24.96 µm, +/-21.45 µm, p< 0.01). Für Optive Fusion® zeigten 3 Corneae Erosiones an Tag 6 (23.11 µm, +/- 37.02 µm, p>0.5), während Vismed Multi® die korneale Heilung nicht negativ beeinflusste. Die BAK Applikation führte zu einer subtotalen Erosion ab Tag 2. Die Glukose und Laktatkonzentrationen blieben unverändert während der Applikationen. Histologisch zeigte sich eine schwere Eipthelopathie sowie Veränderungen des vorderen Hornhautstromas für Optive®. Ähnliche Effekte zeigten sich für Optive Fusion® lediglich in 3 Corneae. Schlussfolgerung: Vismed Multi® Applikationen führt zu einer raschen Heilung. Im Gegensatz zeigt sich für Optive® und weniger ausgeprägt für Optive Fusion® eine kommulative korneale Toxizität. Dies ist am Ehesten auf deren oxidativ wirksame Konservierungsmittel, Purite® zurückzuführen. Diese Daten sollte klinisch überprüft werden, insbesondere um eine Dosis zu ermitteln ab welcher Applikationsfrequenz eine Toxizität auftreten könnte.

Purpose: To analyse the effect of AMS and AMH on HCEC viability, migration and proliferation in vitro. Methods: For AMS preparation, amniotic membrane pieces with a standard size were inserted in a 6-well plate and 5 ml DMEM/F12 (with 5% FBS) per gram tissue was added for 96 hours. After removal of the amniotic membrane, the remaining supernatant was collected for experiments. For AMH preparation, amniotic membranes were first homogenized in liquid nitrogen, then 5 ml DMEM/F12 (with 5% FBS) per gram tissue was added. Following centrifugation, the supernatant was collected for experiments. HCECs were firstly cultured in DMEM/F12 with 5% FBS, 0.5% DMSO, 10 ng/mL human epidermal growth factor, 1% insulin-transferrin-selenium, then were incubated in DMEM/F12 with 5% FBS supplemented by 15%, 30% or 100% AMS or 15% or 30% AMH. Thereafter, HCEC viability was analysed using Cell Proliferation Kit XTT, migration using wound healing assay, proliferation by the cell proliferation ELISA BrdU kit. Results: HCEC viability remained unchanged using 15% or 30% AMS (P=1.0 for both), however, it decreased significantly using 100% AMS (P< 0.001) or 15% (P=0.041) or 30% AMH (P< 0.001), compared to controls. Using 15% or 30% AMS, HCEC migration increased significantly (P< 0.001 for both), compared to controls. Using 15% or 30% AMH (P=0.153; P=0.083), HCEC migration remained unchanged and 100% AMS inhibited HCEC migration (P< 0.001). Fifteen and 30% AMS had no effect on HCEC proliferation (P=0.454 and P=0.119), but 100% AMS (P< 0.001) and 15% (P=0.002) and 30% AMH (P=0.001) inhibited HCEC proliferation significantly, compared to controls. Conclusion: With unchanged HCEC viability and proliferation and increased HCEC migration, 15 and 30% AMS application is the most appropriate method to support epithelial healing. 100% AMS decreases HCEC viability, migration and proliferation. 15 and 30% AMH decreases HCEC viability and proliferation and has no impact on HCEC migration.

Scientific objective: The purpose of the paper lies in improving the effectiveness of treating patients with damaged cornea by means of soft silicon-hydrogel contact lens (CL). Method: There were 11 patients under the supervision, having non-penetrating deep and valve cornea wounds with an apparent corneal syndrome, penetrating cornea wounds: small, wound length up to 3 mm, not sutured air-tightly, also cornea wounds with uneven edges with unsuccessfully completed seal by means of stitches. Against instillations of needed medicinal drugs, medical soft CL of Lotrafilcon A were used in the long-term wearing mode (overnight wearing) within 30 days. Results: by using medical CL, corneal syndrome vanished in observed patients, satisfactory development of cornea wound process was achieved, visual acuity improved and adhesion with a softer scarring was observed. Full recovery was accelerated in average for 5±0.1 days. Conclusions: Medical soft CL of silicon-hydrogel with overnight wearing are one of effective therapeutic methods in treating corneal damages, as well as means of improving patients’ life quality.

Introduction: The therapeutical contact lenses (TCL) are special contact lenses used in the ocular surface diseases. The aim of the study wants to show a possibility to treat penetrating corneal wounds using TCL. Material and method: We took in the study a number of 32 penetrating corneal wounds (with or without suture) in which we used TCL. Results and discussions: In the small penetrating corneal wounds without endoocular membrane issue, we used only TCL. In the large penetrating corneal wounds we used TCL after the wound suture. The results were very good. The benefits of TCL are: minimizing the suture, improving corneal epitelization, tectonic effect and diminishing foreign body sensation. We had no complications. Conclusion: TCL are offering great benefits in treatment of corneal penetrating wounds. TCL are reducing pain, replacing ocular patch, restoring binocularity and improving the quality of life for the patients.

Fragestellung: Im Vergleich zu feucht-warmen Klimazonen gilt die Keratomykose in Europa als eine seltene Erkrankung. Die Fusarium-Arten sind seltene Erreger einer Keratitis und auch in Ländern mit kontinentalem Klima anzutreffen. Im Folgenden sollen die klinischen Verläufe von Patienten mit nachgewiesener Fusariumkeratitis im Zeitraum von Oktober 2012 bis Februar 2015 dargestellt werden. Methode: Retrospektive Auswertung der Behandlungsunterlagen von sechs Patienten mit einer Fusarienkeratitis mit unterschiedlichem Verlauf, behandelt in der Universitätsklinik Regensburg. Ergebnisse: Im angegebenen Zeitraum wurden sechs Fälle einer Fusarienkeratitis bei drei weiblichen und drei männlichen Patienten festgestellt. Alle Patienten waren immunkompetente Erwachsene im Alter zwischen 17 und 92 Jahren (Median 30 Jahre). Bei zwei der Betroffenen war die Reiseanamnese bezüglich des Besuchs von Ländern mit tropischem Klima positiv. Vier Patienten waren Träger von weichen Monatskontaktlinsen. Der Zeitraum bis zur endgültigen Diagnosestellung war im Median 20 Tage. Aufgrund des oberflächlichen Befalls wurden zwei Patienten nur mittels lokaler antimykotischer Therapie behandelt. Bei diesen Patienten zeigte sich ein gutes Ansprechen auf die Therapie und eine gute Visusentwicklung (Durchschnitt 0,2 Log MAR). Bei den restlichen vier Fällen war eine zusätzliche systemische antimykotische Therapie mit Voriconazol 400mg notwendig. Bei allen Patienten wurde eine lokale antimykotische Therapie mit Natamycin Augentropfen (AT) 5% und Voriconazol 1,8% Augentropfen durchgeführt. Bei zwei Patienten waren zusätzlich Vorderkammerspülungen mit Amphotericin B Lösung notwendig. Bei den vier Patienten mit Befall des Hornhautstromas war eine Keratoplastik à chaud erforderlich. Die Therapiedauer betrug im Durchschnitt 4,4 Monate (Median 5 Monate). Bei zwei der operierten Patienten zeigte sich ein stabiler postoperativer Befund und ein guter Endvisus (Durchschnitt 0,3 Log MAR). Eine Patientin wurde extern weiterbehandelt. Bei einem Patienten zeigte sich ein fulminanter Verlauf der Infektion, so dass eine Enukleation erforderlich war. Zusammenfassung: Im Rahmen einer therapieresistenten Keratitis sollte eine Fusarieninfektion in Betracht gezogen werden. In dieser Fallserie wurde gezeigt wie unterschiedlich der Verlauf einer Fusarieninfektion sein kann. Diese ist nicht immer kontaklinsenassoziiert und kommt auch in Ländern mit kontinentalem Klima vor.

Ziel: Die Anwendung von autologen Serumaugentropfen (AS) kann eine alternative Behandlungsmethode für therapieresistente korneale Epitheldefekte darstellen. Bei persistierenden Epitheldefekten, könnten durch die im Hornhautstroma anwesenden Keratozyten produzierten Zytokinein der Epithelwundheilung eine entscheidende Rolle spielen. Ziel dieser Studie ist es, den Effekt von AS auf Sekretion von Fibroblast growth factor basic (FGFb), Hepatocyte growth factor (HGF), Keratinocyte growth factor (KGF) und Transforming growth factor β1 (TGF-β1) von Keratozyten in vitro zu untersuchen. Methode: Autologe Serumaugentropfen von 5 Patienten wurden nach der Standardmethode der LIONS Hornhaut Bank Saar-Lor-Lux präpariert und bei -80°C tiefgefroren. Primäre humane Keratozyten wurden durch enzymatische Behandlung mit Collagenase A (1mg/ml) aus humanen Korneoskleralscheiben isoliert (n=5) und in DMEM/Ham´s Kulturmedium, versetzt mit 10% fetalem Kälberserum kultiviert. Für den Testansatz wurden die Keratozyten mit 15 oder 30% AS inkubiert und nach 24h die Sekretion von FGFb, HGF, KGF und TGFβ1 mit einem Enzyme linked immunoabsorbent assay (ELISA) aus dem Kulturüberstand photometrisch analysiert. Der Einfluss der AS Konzentrationen von unterschiedlichen Patienten auf die Sekretion von FGFb, HGF, KGF und TGFβ1 von Keratozyten verschiedener Spender wurde mit einem verallgemeinerten linearen Modell (Generalized Estimating Model, GEM) analysiert. Ergebnisse: Die Konzentrationen von FGFb, HGF, KGF und TGFβ1 im Kulturüberstand der Keratozyten waren heterogen (11,83±38,76/ 8779,12±7537,56/ 36,06±81,65/ 1533,12±987,06 pg/µg Protein). Ein signifikanter Einfluss von AS Konzentrationen und Herkunft auf die FGFb, HGF, KGF und TGFβ Sekretion von Keratozyten konnte nicht nachgewiesen werden (p>0,11). Schlussfolgerung: 15 und 30% autologes Serum hat keinen Einfluss auf die Sekretion von Wachstumsfaktoren von Keratozyten in vitro. Weitere Studien mit einer höheren Anzahl von Patienten sollten zur Unterstützung unserer Ergebnisse durchgeführt werden.

Aim: To analyse the results of the therapeutic keratoplasty (KP), performed in the department of corneal pathology in patients with severe inflammatory diseases of the cornea in 2013-2014. Methods: Retrospective analyzed results of 189 therapeutic KP in 187 patients over period 2 years between January 2013 to December 2014. The mean age of patients was 48.5±16.7 years, male - 126 (66,7%) female - 63 (33,3 %). Indications for keratoplasty were: severe keratitis – 18,6%, corneal ulcers – 74,3%, corneal abscess - 7,1%. Etiology of inflammatory process was : bacterial - 27, 5%, herpetic - 26,0%, fungal - 6,3%, mixed infection - 9,5%, neurotrophic - 14,8%, autoimmune - 15,9%. Depending on the severity of the initial state we have performed: biological covering by Putchkovskaja (n=16), superficial or deep lamellar KP (n=96), penetrating classical KP (n=22) and step-penetrating KP (n=41), mushroom KP (n = 5), KP with two grafts (n=7), exchange KP (n=2). In five cases additionally was performed partial blefarorrafy. The criteria of treatment results were: eye saving, stopping of the inflammatory process, presence of complications, the level of intraocular pressure (IOP), visual acuity, the perspectives for the surgery with optical purpose in the future. Results: The therapeutic effect was achieved in 186 (98,3%) patients, in 3 cases was made evisceration. Clear corneal graft after lamellar KP was in 70 eyes (72,9%), after penetrating KP in 39 (55,7%). Postoperative complication were: graft rejection 67 (35,4%), among them 26 finally failed; cataracta progression - 54 (28,5%); persistent epithelial defects of the graft - 36 (19,1%); reccurens of main diseases 27 (14,2%); secondary glaucoma in 72 cases (38,1%). In the mean follow-up period 16,7±5,8 months were performed antiglaucomatous operations at 45 patients (23,8%), cataracta extraction with IOL implantation – 32 (16,9%), regrafts – 27 (14,3%), triple procedure – 9 (4,6%). The mean visual acuity after treatment was 0,25± SD0,12. Conclusion: Therapeutic KP is an effective method of rehabilitation of patients with severe inflammatory diseases of the cornea, it allows to remove the affected layers of the cornea, excise destructive corneal tissue, to stop the inflammatory process and save the perspective of surgical treatment with an optical aim.

Purpose: To date, studies focusing on the glycosylation pattern of the human tear film proteome are still rare and limited on single or function-related proteins. However, the structure and stability of proteins are highly influenced by these modifications leading to changes of their properties like binding or transport functions. For this reason, the primary objective of this study was to provide the basic fundamental information about the glycoproteins in the human tear film. Methods: Tear samples were collected from 8 healthy volunteers (mean age: 28 ± 3) by capillary technique and Schirmer’s strips. The samples were pooled and two-dimensional gel electrophoresis (2-DE) was performed. The gels were stained with two different dyes: Colloidal Blue Staining kit to visualize the total protein amount and Glycoprotein staining kit to detect glycoproteins. The protein spots were analyzed by proteomic based liquid chromatography – mass spectrometry (LC-MS) approaches. State-of-the-art bioinformatic tools based on de novo sequencing strategy were used to detect glycoproteins and to characterize their glycosylation sites. Results: By 2-DE analysis complex glycosylation profiles of the human tear film proteome could be identified. By means of bottom-up LC-MS analysis, up to 30 proteins (false discovery rate < 1%) were detected with distinct glycosylation characteristics. Furthermore, it was possible to identify these glycoproteins in tear samples collected independently by two different approaches. State-of-the-art bioinformatics tools were instrumental in identifying several new glycosylation sites in both high abundant proteins (e.g. Lactotransferrin or Immunoglobulin alpha-1 chain) as well as in low abundant proteins (e.g. Cadherin-6 or Zymogen granule protein 16 homolog B). Conclusion: In this study it could be clearly demonstrated that human tear film proteins show a versatile and complex glycosylation profile. For many diseases, amongst others autoimmune disorders, it is already known that aberrant glycosylations of proteins are involved in various pathophysiological processes, promoting the concept that glycosylation patterns could be an attractive target for biomarker research. Further studies are of great importance to investigate the complex modification pattern of the human tear film proteome for a better understanding how glycosylations contribute to the development of eye diseases.