Pathophysiology of aniridia-associated keratopathy: Developmental aspects and unanswered questions.

Aniridia, a rare congenital disease, is often characterized by a progressive, pronounced limbal insufficiency and ocular surface pathology termed aniridia-associated keratopathy (AAK). Due to the characteristics of AAK and its bilateral nature, clinical management is challenging and complicated by the multiple coexisting ocular and systemic morbidities in aniridia. Although it is primarily assumed that AAK originates from a congenital limbal stem cell deficiency, in recent years AAK and its pathogenesis has been questioned in the light of new evidence and a refined understanding of ocular development and the biology of limbal stem cells (LSCs) and their niche. Here, by consolidating and comparing the latest clinical and preclinical evidence, we discuss key unanswered questions regarding ocular developmental aspects crucial to AAK. We also highlight hypotheses on the potential role of LSCs and the ocular surface microenvironment in AAK. The insights thus gained lead to a greater appreciation for the role of developmental and cellular processes in the emergence of AAK. They also highlight areas for future research to enable a deeper understanding of aniridia, and thereby the potential to develop new treatments for this rare but blinding ocular surface disease.

[Stage-appropriate treatment of keratoconus]. / Stadiengerechte Therapie des Keratokonus.

Keratoconus (KC) is a progressive cone-shaped corneal protrusion that causes paracentral thinning at the apex of the cone and typically occurs asymmetrically on both sides. After a careful anamnesis and classification of the degree of severity a targeted treatment appropriate to the stage of the disease is available. If the visual acuity is no longer sufficient, rigid gas-permeable contact lenses (CL) are fitted by a specialist. Riboflavin UVA cross-linking (CXL) is recommended in cases of progression and visual acuity that is still useful for the patient. Intracorneal ring segments (ICRS) are indicated for CL intolerance in cases of reduced visual acuity and a clear central cornea. If the stage is more advanced, deep anterior lamellar keratoplasty (DALK) or penetrating keratoplasty (PKP) is recommended. A PKP is contraindicated in acute KC but deep stromal sutures for readaptation of the Descemet tear with gas filling of the anterior chamber can considerably shorten the course. Almost no other eye disease is nowadays as easily accessible for an early instrument-based diagnosis and stage-appropriate treatment as KC.

Abnormal neovascular and proliferative conjunctival phenotype in limbal stem cell deficiency is associated with altered microRNA and gene expression modulated by PAX6 mutational status in congenital aniridia.

PURPOSE: To evaluate conjunctival cell microRNA (miRNAs) and mRNA expression in relation to observed phenotype of progressive limbal stem cell deficiency in a cohort of subjects with congenital aniridia with known genetic status. METHODS: Using impression cytology, bulbar conjunctival cells were sampled from 20 subjects with congenital aniridia and 20 age and sex-matched healthy control subjects. RNA was extracted and miRNA and mRNA analyses were performed using microarrays. Results were related to severity of keratopathy and genetic cause of aniridia. RESULTS: Of 2549 miRNAs, 21 were differentially expressed in aniridia relative to controls (fold change ≤ -1.5 or ≥ +1.5). Among these miR-204-5p, an inhibitor of corneal neovascularization, was downregulated 26.8-fold in severely vascularized corneas. At the mRNA level, 539 transcripts were differentially expressed (fold change ≤ -2 or ≥ +2), among these FOSB and FOS were upregulated 17.5 and 9.7-fold respectively, and JUN by 2.9-fold, all being components of the AP-1 transcription factor complex. Pathway analysis revealed enrichment of PI3K-Akt, MAPK, and Ras signaling pathways in aniridia. For several miRNAs and transcripts regulating retinoic acid metabolism, expression levels correlated with keratopathy severity and genetic status. CONCLUSION: Strong dysregulation of key factors at the miRNA and mRNA level suggests that the conjunctiva in aniridia is abnormally maintained in a pro-angiogenic and proliferative state, and these changes are expressed in a PAX6 mutation-dependent manner. Additionally, retinoic acid metabolism is disrupted in severe, but not mild forms of the limbal stem cell deficiency in aniridia.

Asymmetry between Left and Right Eyes in Keratoconus Patients Increases with the Severity of the Worse Eye.

PURPOSE: To evaluate whether the inter-eye asymmetry of keratoconus (KC) patients is different from a healthy control group and to investigate how asymmetry changes with increasing severity of the disease. METHODS: In this retrospective study, we included both eyes of 350 patients with KC (age 35 ± 13 years) and 68 candidates planned for refractive surgery (control group, age 37 ± 11 years). Inclusion criteria for the KC group were keratoconus in at least one eye with Pentacam Topographical Keratoconus Classification (TKC) of at least 0.5. Patients eligible for refractive surgery in both eyes were included in the control group. Corneal tomography as well as Ocular Response Analyzer measurements were compared between both groups. Subgroup analysis was performed with respect to the TKC staging. Asymmetry was provided as worse eye (defined by higher TKC) minus fellow eye. RESULTS: In the KC group, both eyes showed the same TKC staging in 30.6%, a difference of one stage in 34.0% and of two stages in 24.6% of the patients. The inter-eye asymmetry in the keratoconus group was significantly larger than that in the control group. Corneal power showed an asymmetry of 3.8 ± 4.0 D in keratoconus eyes versus 0.22 ± 0.17 D in the control group. Central corneal thickness (CCT) asymmetry was 34 ± 30 µm versus 6 ± 5 µm, respectively. The Keratoconus Match Index showed an asymmetry of 0.40 ± 0.35 versus 0.15 ± 0.14. The difference between both eyes increased with increasing TKC of the worse eye. CONCLUSIONS: Inter-eye asymmetry is larger in keratoconus than in normal eyes, and it increases with keratoconus severity in the worse eye.

[Safety of Hyaluronic Acid in Postoperative Treatment after Penetrating Keratoplasty]. / Verträglichkeit von Hyaluronsäure zur Oberflächenpflege nach perforierender Keratoplastik.

BACKGROUND: The objective of this clinical trial was to confirm the safety of phosphate-free high molecular weight hyaluronate (HYLO®-GEL) as lubricant for postoperative care after penetrating keratoplasty. PATIENTS AND METHODS: 51 eyes were followed up 6 weeks after penetrating keratoplasty under therapy with phosphate-free high molecular weight hyaluronate. Visual acuity, intraocular pressure, conjunctival hyperaemia, conjunctival chemosis, corneal fluorescein staining, corneal infiltrates, corneal endothelial-epithelial decompensation, Descemet's folds, Tyndall effect, cells in the anterior chamber and subjective assessment of lacrimation, foreign body sensation, pressure sensation, burning, mucus and itching were recorded on the first, third and fifth postoperative days and six weeks postoperatively. Tolerance was rated by the patient and the attending ophthalmologist. RESULTS: The following objective parameters improved significantly from day 1 to week 6 in patients treated with phosphate-free high molecular weight hyaluronate: visual acuity, conjunctival hyperaemia, conjunctival chemosis, corneal fluorescein-staining, corneal endothelial-epithelial decompensation, Descemet's folds, the maximum diameter of corneal erosion, Tyndall effect and cells in the anterior chamber. In addition, there was significant improvement in the course of the following subjective complaints: lacrimation, foreign body sensation, pressure sensation and itching. In total 63 % (32/47) of the study patients reported excellent local tolerance of phosphate-free high molecular weight hyaluronate on day one postoperatively. This proportion rose to 86 % (37/43) 6 weeks postoperatively. The safety of phosphate-free high molecular weight hyaluronate was rated by the ophthalmologists as "perfect" in 92 %, as "acceptable" in 6 % and as "unacceptable" in 2 % of eyes. CONCLUSION: Therapy with phosphate-free high molecular weight hyaluronate results in significant improvements in objective findings and subjective symptoms after penetrating keratoplasty. Tolerance was rated as "excellent" by the ophthalmologist in 92 % of patients in the early postoperative follow-up.

Anatomy-based DMEK Wetlab in Homburg/Saar: Novel aspects of donor preparation and host maneuvers to teach descemet membrane endothelial keratoplasty.

Use of Descemet Membrane Endothelial Keratoplasty (DMEK) has been limited because of problems with donor preparation, i.e. tearing of the Descemet membrane and difficulties in unfolding the Endothelium-Descemet-Membrane-Layer (EDML) in the anterior chamber (AC). The purpose of this work was to describe a novel approach to teaching anatomy-based donor and recipient preparation in a DMEK-Wetlab. We teach successful mono-manual donor preparation of human corneas in organ culture not suitable for transplantation, including peripheral markers for orientation. We also teach safe recipient preparation in a freshly-enucleated pig eye in organ culture preservation medium for atraumatic introduction of the EDML roll into the AC, reliable orientation of the EDML during surgery, and stepwise unfolding within the AC. Twenty-two candidates in the 1. Homburg Cornea Curriculum HCC 2015 who practiced both preparations using three human donor corneas and three pig eyes assessed the procedure as follows: (1) overall grade of the Wetlab 1.4 (median 1, range 1 to 2 - on a scale from 1 (excellent) to 6 (terrible); (2) most participants and tutors stated that the Wetlab is most effective for colleagues who have some previous experience with corneal microsurgery. Our novel anatomy-based approach to simulating donor preparation and graft implantation for DMEK seems to meet the expectations and requirements of colleagues with previous experience in corneal microsurgery and will help to reduce the rate of complications for incipient DMEK surgeons in the future. Clin. Anat. 31:16-27, 2018. © 2017 Wiley Periodicals, Inc.

[Amniotic Membrane Suspension and Autologous Serum - Are they Important for Wound Healing?] / Amnionmembransuspension und autologes Serum ­ spielt der Inhalt eine Rolle für die Wundheilung?

Autologous serum (AS) and amniotic membrane transplantation (AMT) are used in the treatment of several ocular surface diseases. AMT is associated with a surgical intervention. Surgery could be avoided by using eye drops prepared from an amniotic membrane homogenate (AMH) or amniotic membrane suspension (AMS). EGF, bFGF, IL-6 and IL-8 were detected in AMS. However, EGF and bFGF concentrations in AMS were about 1.7-17× lower than in AMH, and IL-6 and IL-8 could not be detected in AMH. 100 % AMS, 15 and 30 % AMH significantly decreased proliferation of human corneal epithelial cells (HCECs) compared to controls (p = <0.002 for all), but 15 and 30 % AMS did not affect proliferation. Migration increased significantly compared to controls with 15 and 30 % AMS (p < 0.001), but did not change significantly with 15 or 30 % AMH (p = 0.153 and p = 0.083). Proliferation of HCECs was significantly greater with 15 % AS than with 30 % AS (p < 0.001). HCEC migration was significantly greater with 30 % AS than with 5 % AS (p < 0.01). In summary, 15 and 30 % AMS and 15 and 30 % AS exhibit the best supportive effect on human corneal epithelial cells. Nevertheless, we always have to keep in mind that individual growth factor concentrations exhibit high inter-individual fluctuations in AS or AMS eye drops.

[Functional Results and Graft Failure after Repeat Keratoplasty]. / Funktionelle Ergebnisse und Transplantatversagen nach Re-Keratoplastik.

Background and Purpose Keratoplasty is the oldest and most successful transplantation method in man. Despite the immunological privilege of the cornea, the graft fails in up to 10 % of patients, depending on corneal disease, quality of donor tissue and patient compliance, and repeat keratoplasty may then be necessary. The aim of this study was to verify factors which affect the functional result and graft survival after repeat keratoplasty. Patients and Methods In this single centre, consecutive retrospective case series, patients were included who were treated at Department of Ophthalmology, Saarland University Medical Centre with repeat keratoplasty between January 1st 2001 and December 31st 2010. Data were recorded with an Access Database from the medical records and statistically analysed with SPSS 19.0. The surgical reports and the results from the follow-up examinations concentrated on visual acuity, intraocular pressure, endothelial cell density, corneal topography and tomography and corneal graft reactions. Results The results of the study showed that trephination with the excimer laser was significantly superior (p = 0.009) to mechanical trephination with a motor trephine with respect to corrected visual acuity. Corrected visual acuity after 2 years was 0.39 in the laser group, compared to 0.16 in the group with mechanical trephination. A graft diameter under 7.5 mm showed a significant negative impact on postkeratoplasty astigmatism (p = 0.004). After a follow-up period of 120 months, the rate of immunological graft rejection was 20/60 % (p = 0.259) in the patient group with graft diameters ≤ 7.5 mm/> 7.5 mm, respectively. Conclusion The present study shows that repeat keratoplasty using excimer laser gives better functional rehabilitation, if corneal morphology permits this (for example, the vascularisation). However, the trephination method has no significant impact on graft survival. The basic principle for repeat keratoplasties is that the diameter of the graft should be as great as possible but as small as necessary.

[PKP for Keratoconus - From Hand/Motor Trephine to Excimer Laser and Back to Femtosecond Laser]. / Perforierende Keratoplastik bei fortgeschrittenem Keratokonus - vom Hand-/Motortrepan hinauf zum Excimerlaser und zurück zum Femtosekundenlaser.

For patients with keratoconus, rigid gas-permeable contact lenses are the first line correction method and allow good visual acuity for quite some time. In severe stages of the disease with major cone-shaped protrusion of the cornea, even specially designed keratoconus contact lenses are no longer tolerated. If there are contraindications for intrastromal ring segments, corneal transplantation typically has a very good prognosis. In patients with advanced keratoconus - especially after corneal hydrops due to rupture of Descemet's membrane - penetrating keratoplasty (PKP) is still the first line surgical method. Non-contact excimer laser trephination seems to be especially beneficial for eyes with iatrogenic keratectasia after LASIK and for patients with repeat grafts due to "keratoconus recurrences" due to small grafts with thin host cornea. For donor trephination from the epithelial side, an artificial chamber is used. Wound closure is achieved with a double running cross-stitch suture according to Hoffmann. Graft size is adapted individually, depending on corneal size ("as large as possible - as small as necessary"). Limbal centration is preferred intraoperatively, due to optical displacement of the pupil. During the last 10 years, femtosecond laser trephination has been introduced from the USA as a potentially advantageous approach. Prospective clinical studies have shown that the technique of non-contact excimer laser PKP improves donor and recipient centration, reduces "vertical tilt" and "horizontal torsion" of the graft in the recipient bed, and thus results in significantly less "all-sutures-out" keratometric astigmatism (2.8 vs. 5.7 D), more regular topography (surface regularity index [SRI] 0.80 vs. 1.0) and better visual acuity (0.80 vs. 0.60), in comparison to the motor trephine. The stage of the disease does not influence functional outcome after excimer laser PKP. However, the refractive outcome of femtosecond laser keratoplasty resembles that with motor trephine. In contrast to the undisputed clinical advantages of excimer laser keratoplasty with orientation teeth/notches in keratoconus, the major disadvantage of femtosecond laser application is still the necessity of suction and applanation of the cone during trephination, with intraoperative disadvantages and high postoperative astigmatism.

Crosslinking and corneal cryotherapy in acanthamoeba keratitis -- a histological study.

PURPOSE: Acanthamoeba keratitis is rare, but difficult to treat. Penetrating keratoplasty is performed in therapy-resistant cases. Nevertheless, subsequent recurrences occur in 40 % of the cases. In addition to triple-topical therapy (polyhexamid, propamidinisoethionat, neomycin), treatment alternatives are corneal cryotherapy and/or crosslinking (CXL). The aim of our present histological study was to analyze the persistence of acanthamoebatrophozoites and cysts, the persistence of bacteria, and activation of keratocytes in corneas of acanthamoeba keratitis patients following corneal cryotherapy and/or CXL. PATIENTS AND METHODS: We analyzed histologically corneal buttons (from penetrating keratoplasties) of nine patients with acanthamoeba keratitis, following corneal cryotherapy (two patients) or a combination of crosslinking and corneal cryotherapy (seven patients), using haematoxilin­eosin, periodic acid Schiff (PAS), Gram and alpha-smooth muscle actin (alpha-SMA) stainings. RESULTS: Acanthamoeba trophozoites persisted in three corneas after cryotherapy and CXL. Cysts persisted in one of two corneas following corneal cryotherapy and in six of seven corneas after a combination of CXL and cryotherapy. One cornea showed positive Gram staining, but there were no alpha-SMA positive keratocytes in any of the corneas. CONCLUSIONS: Crosslinking and corneal cryotherapy have only limited impact on killing of acanthamoeba trophozoites, cysts, or bacteria. Corneal cryotherapy and CXL did not stimulate myofibroblastic transformation of keratocytes.

[Reproducibility of Scheimpflug Tomography Measurements Regarding Corneal Front and Back Surface Power]. / Reproduzierbarkeit der Messungen der Scheimpflug-Tomografie für die Brechwerte der vorderen und hinteren Hornhautgrenzfläche.

BACKGROUND AND PURPOSE: The Oculus Pentacam® is one of the most commonly used devices in ophthalmology for assessment of the anterior eye segment. The purpose of this study was to determine the reproducibility of the corneal power as measured by the Pentacam® in a normal population. METHODS: We enrolled 25 eyes of 25 subjects aged between 13 and 68 (46.7 ± 21.7) years, within a spherical equivalent from - 4 D to + 4 D and refractive cylinder up to 5 D and without notable pathologies or history of surgery. A sequence of 5 measurements was performed using the Pentacam®, the Zeiss IOLMaster® as well as an autorefractometer after (re-)positioning the patient's head and the measurement device. From the Pentacam® we collected power data for the corneal front and back surfaces, apical corneal thickness at apex and internal anterior chamber depth. From the IOLMaster®, we extracted corneal power data, and from the autorefractometer we obtained refractive power data of both cardinal meridians. For statistical analysis, we used Cronbach's α as a measure of reproducibility and Spearman's rank correlation test for correlation of data. RESULTS: Pentacam® yields highly reproducible power data for both corneal surfaces, with Cronbach's α of greater than 0.97 for primary parameters (e.g. radii of curvature) and 0.9 for secondary parameters (e.g. vector components of astigmatism). Central corneal thickness and eccentricities of both surfaces as well as anterior chamber depth yielded highly reproducible values, with α of ≥ 0.97. Corneal power derived from IOLMaster® and objective refraction extracted from the autorefractometer yielded α values of around 0.9. Mean corneal power of Pentacam® and IOLMaster® correlated well, but there was no correlation to spherical equivalent. Astigmatism values from Pentacam® and IOLMaster® correlated well with each other and with refractive cylinder. CONCLUSION: Our results indicate that measurements of anterior segment using the Pentacam® yield highly reproducible results in a normal population without major pathologies of the anterior eye segment.

[Effect of Photodynamic Inactivation (PDI) using Riboflavin-Conjugated Antibody against Staphylococcus aureus]. / Effekt photodynamischer Inaktivierung (PDI) mit Riboflavin oder riboflavinkonjugierten Antikörpern gegen Staphylococcus aureus.

PURPOSE: Crosslinking/riboflavin-UVA photodynamic therapy is a potential treatment alternative in antibiotic resistant infectious keratitis. For photodynamic therapy a specific (against bacteria) conjugated antibody may be used in order to increase the effect of the treatment. In our present study we analysed the impact of photodynamic inactivation using riboflavin-conjugated antibody or riboflavin alone on Staphylococcus aureus, in vitro. METHODS: Staphylococcus aureus (S. aureus) was incubated in 1 : 100 diluted riboflavin-conjugated antibody (R-AB) for 30 minutes in darkness. Following UVA-light illumination (375 nm) with an energy dose of 2, 3, 4 and 8 J/cm(2), bacteria were brought to blood agar Plates for 24 hours before colony-forming unit (CFU) counting. In an additional group, we incubated bacteria to 0, 0.05 or 0.1 % riboflavin 5-phosphate as described above followed by illumination using UVA light (375 nm) with an energy dose of 2 J/cm(2), before CFU counting. RESULTS: The number of CFU decreased significantly (inactivation of 36 %, p = 0.022) using 1 : 100 diluted riboflavin-conjugated antibody and 2 J/cm(2) UVA-light illumination, compared to untreated controls. The use of 3, 4 und 8 J/cm(2) energy dose and R-AB in 1 : 100 dilution did not further change the decrease of CFU (inactivation of 39, 39 and 40 %; p = 0.016; p = 0.016; p = 0.015). The use of 0.05 % or 0.1 % riboflavin 5-phosphate alone and UVA-light illumination reduced the CFU count significantly (inactivation of 73 and 55 %; p = 0.002; p = 0.005), compared to untreated controls. CONCLUSIONS: The use of riboflavin-conjugated antibody or 0.05 % or 0.1 % riboflavin 5-phosphate and UVA-light illumination reduces the number of CFU of S. aureus. However, none of these photodynamic therapies reached the necessary 99 % killing rate of these bacteria. Further work is needed to increase the efficacy of riboflavin-conjugated antibodies against antibiotic resistant bacteria.

[Surgeon's Impact on Graft Endothelial Cell Density and Corneal Thickness after Penetrating Keratoplasty for Keratoconus and Fuchs' Dystrophy]. / Einfluss der individuellen mikrochirurgischen Technik auf Endothelzelldichte und Pachymetrie nach perforierender Keratoplastik bei Patienten mit Fuchs-Dystrophie oder Keratokonus.

PURPOSE: The aim of this study was to examine the short- and long-term variability of graft endothelial cell density (ECD) and central corneal thickness (CCT) after penetrating keratoplasty (PKP) between different surgeons. PATIENTS AND METHODS: 370 eyes with keratoconus (KC) or Fuchs' dystrophy (FUCHS) after standardised primary excimer laser PKP (four experienced surgeons) were analysed. ECD and CCT were determined at 12 months after PKP before suture removal, and at two years after final suture removal. RESULTS: 12 months after PKP, ECD was significantly higher for surgeon 1 in FUCHS as compared to surgeons 3 (p = 0.05) and 4 (p = 0.04), in KC it was significantly higher for surgeon 1 as compared to surgeon 2 (p = 0.001) and for surgeon 1 as compared to surgeon 4 (p = 0.006). However, in FUCHS this difference was no longer significant two years after suture removal (p > 0.5), and in KK only in comparison of surgeons 1 and 2 (p = 0.04). RESULTS concerning CCT were inhomogeneous. CONCLUSIONS: The surgeon's individual handwriting seems to have an impact on the ECD in the short-term after PKP. However, differences between surgeons tend to become insignificant in the long run after suture removal.

[The German Acanthamoeba keratitis register: Initial results of a multicenter study]. / Das Deutsche Akanthamöbenkeratitis-Register : Erste Ergebnisse einer multizentrischen Erhebung.

BACKGROUND AND PURPOSE: In September 2011 the cornea section of the German Ophthalmological Society (DOG) established the first German Acanthamoeba keratitis registry. The data of this multicenter survey are being collected, compiled and evaluated at the Department of Ophthalmology at the Saarland University. The aim of this article is to present an intermediate report. PATIENTS AND METHODS: Data from 172 eyes with Acanthamoeba keratitis were collected during the last 10 years. For this interim report we actually evaluated 121 eyes (60.2 % female patients, average age 41.3 years) and collected the following data: date of onset of symptoms, date and method of diagnosis, initial diagnosis, anamnestic data, clinical symptoms and signs at diagnosis and during follow-up, conservative and surgical therapy. Criteria for inclusion in the Acanthamoeba registry was the established diagnosis of an Acanthamoeba keratitis with at least one of the methods described in this article. RESULTS: Acanthamoeba keratitis could be histologically proven in 55.3 % of the cases, via PCR in 25.6 %, with confocal microscopy in 20.4 % and using in vitro cultivation in 15.5 %. Clinical symptoms and signs in Acanthamoeba keratitis were pain in 67.0 %, ring infiltrates in 53.4 %, pseudodendritiform epitheliopathy in 11.7 % and keratoneuritis in 5.8 %. In 47.6 % of the cases the initial diagnosis was herpes simplex virus keratitis followed by bacterial keratitis in 25.2 % and fungal keratitis in 3.9 %. Acanthamoeba keratitis was the correct initial diagnosis in only 23.2 % of cases. The average time period between first symptoms and diagnosis was 2.8 ± 4.0 months (range 0-23 months). A triple therapy with Brolene® Lavasept® and antibiotic eye drops at least 5 ×/day was used in 54.5 % of eyes (n = 66). Penetrating keratoplasty was performed in 40.4 %, in 18 cases in combination with cryotherapy of the cornea. The mean graft diameter was 7.9 ± 1.1 mm (range 3.5-11.0 mm). The final visual acuity (Snellen visual acuity chart at 5 m) was comparable in the two groups of eyes with (5/40 ± 5/25) and without (5/32 ± 5/25) keratoplasty. CONCLUSION: Acanthamoeba keratitis is a rare and often very late diagnosed disease and two thirds of the cases were initially misdiagnosed. The early recognition of the typical symptoms is crucial for the prognosis of the disease. All ophthalmological departments in Germany are invited to submit further data of all confirmed cases (berthold.seitz@uks.eu), whether retrospectively or prospectively in order to generate an adequate standardized diagnostic and therapeutic approach for this potentially devastating disease.

[Growth Factors and Interleukins in Amniotic Membrane Tissue Homogenate]. / Wachstumsfaktoren und Interleukine in Amniongewebehomogenat.

PURPOSE: Application of amniotic membrane homogenate eye drops may be a potential treatment alternative for therapy resistant corneal epithelial defects. The purpose of this study was to determine the concentrations of epidermal growth factor (EGF), fibroblast growth factor basic (bFGF), hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), interleukin-6 (IL-6) and interleukin-8 (IL-8) in amniotic membrane homogenates. METHODS: Amniotic membranes of 8 placentas were prepared and thereafter stored at - 80 °C using the standard methods of the LIONS Cornea Bank Saar-Lor-Lux, Trier/Westpfalz. Following defreezing, amniotic membranes were cut in two pieces and homogenized in liquid nitrogen. One part of the homogenate was prepared in cell-lysis buffer, the other part was prepared in PBS. The tissue homogenates were stored at - 20 °C until enzyme-linked immunosorbent assay (ELISA) analysis for EGF, bFGF, HGF, KGF, IL-6 and IL-8 concentrations. RESULTS: Concentrations of KGF, IL-6 and IL-8 were below the detection limit using both preparation techniques. The EGF concentration in tissue homogenates treated with cell-lysis buffer (2412 pg/g tissue) was not significantly different compared to that of tissue homogenates treated with PBS (1586 pg/g tissue, p = 0.72). bFGF release was also not significantly different using cell-lysis buffer (3606 pg/g tissue) or PBS treated tissue homogenates (4649 pg/g tissue, p = 0.35). HGF release was significantly lower using cell-lysis buffer (23,555 pg/g tissue), compared to PBS treated tissue (47,766 pg/g tissue, p = 0.007). CONCLUSION: Containing EGF, bFGF and HGF, and lacking IL-6 and IL-8, the application of amniotic membrane homogenate eye drops may be a potential treatment alternative for therapy-resistant corneal epithelial defects.

[Interobserver and intraobserver reliability of corneal surface temperature measurements with the TG-1000 thermograph in normal eyes]. / Inter- und Intraobserverreliabilität der kornealen Oberflächentemperaturmessung mit dem TG-1000 bei Normalaugen.

PURPOSE: The aim of this study was to analyze the reliability of temperature measurements with the ocular TG-1000 thermograph in a setup of sequential measurements performed by one observer (intraobserver) and a sequence of measurements performed by different observers (interobserver) in normal subjects without pathologies of the anterior segment of the eye. PATIENTS AND METHODS: A total of 50 right eyes from 50 individuals (mean age 29.1 ± 7.9 years) without ocular pathologies or history of ocular surgery were enrolled in this prospective monocentric clinical case series. Eyes with signs of dry eye syndrome (based on a positive McMonnies questionnaire) were excluded from the study. Corneal surface temperature measurements were performed by three examiners to assess interobserver reliability. In addition, in a subgroup of 22 individuals, a sequence of 3 measurements were performed by 1 of the examiners to examine intraobserver reliability. Corneal surface temperature was measured within an interval of 10 s (11 frames) on a region of interest of 16 ± 12 mm (320 ± 240 pixels). Central and mid-peripheral local temperatures at 3 mm (3, 6, 9 and 12 o'clock) were extracted and analyzed from the raw data. RESULTS: The ocular TG-1000 thermograph yielded consistent results for the interobserver as well as intraobserver conditions in measuring corneal surface temperature in the center as well as mid-periphery of the cornea. Cronbach's alpha was 0.9 or higher at all corneal locations, which proves a high consistency of results for the interobserver and intraobserver measurements. The average corneal surface temperature ranged between 34.0 °C and 34.7 °C with a slight decrease from the upper temporal (9 and 12 o'clock) to the lower nasal (3 and 6 o'clock) quadrants. CONCLUSION: The TG-1000 thermograph yielded consistent results of corneal surface temperature in individuals without anterior segment pathologies or history of ocular surgery. With the option of raw data export (11 frames within 10 s with a lateral resolution of 320 × 240 pixels) the thermograph offers a wide range of new diagnostic options for a spatiotemporal analysis of corneal surface temperature.