Christina Jacobi

Corneal Limbal Microenvironment Can Induce Transdifferentiation of Hair Follicle Stem Cells into Corneal Epithelial‐like Cells

The aim of this study was to investigate the transdifferentiation potential of murine vibrissa hair follicle (HF) stem cells into corneal epithelial‐like cells through modulation by corneal‐ or limbus‐specific microenvironmental factors. Adult epithelial stem cells were isolated from the HF bulge region by mechanical dissection or fluorescence‐activated cell sorting using antibodies to α6 integrin, enriched by clonal expansion, and subcultivated on various extracellular matrices (type IV collagen, laminin‐1, laminin‐5, fibronectin) and in different conditioned media derived from central and peripheral corneal fibroblasts, limbal stromal fibroblasts, and 3T3 fibroblasts. Cellular phenotype and differentiation were evaluated by light and electron microscopy, real‐time reverse transcription‐polymerase chain reaction, immunocytochemistry, and Western blotting, using antibodies against putative stem cell markers (K15, α6 integrin) and differentiation markers characteristic for corneal epithelium (K12, Pax6) or epidermis (K10). Using laminin‐5, a major component of the corneo‐limbal basement membrane zone, and conditioned medium from limbal stromal fibroblasts, clonally enriched HF stem and progenitor cells adhered rapidly and formed regularly arranged stratified cell sheets. Conditioned medium derived from limbal fibroblasts markedly upregulated expression of cornea‐specific K12 and Pax6 on the mRNA and protein level, whereas expression of the epidermal keratinocyte marker K10 was strongly downregulated. These findings suggest that adult HF epithelial stem cells are capable of differentiating into corneal epithelial‐like cells in vitro when exposed to a limbus‐specific microenvironment. Therefore, the HF may be an easily accessible alternative therapeutic source of autologous adult stem cells for replacement of the corneal epithelium and restoration of visual function in patients with ocular surface disorders. STEM CELLS 2009;27:642–652

Tear film osmolarity measurements in dry eye disease using electrical impedance technology.

Purpose: Tear film hyperosmolarity is recognized as an important pathogenetic factor in dry eye syndrome, but difficulties in its measurement have limited its utility in the recent past. This prospective, nonrandomized, clinical single-center study investigates the osmolarity in tear samples of patients with keratoconjunctivitis sicca compared with healthy controls. Methods: One hundred thirty-three patients [aged 58 years (51–64 years), 86 women and 47 men] with moderate to severe keratoconjunctivitis sicca and 95 controls [aged 52 years (48–61 years), 55 women and 40 men] were enrolled in the trial. Tear samples were collected directly from the inferior lateral tear meniscus. Inclusion criteria were a tear breakup time of less than 5 seconds, a Schirmer test with anesthesia less than 5 mm, and positive symptoms (Ocular Surface Disease Index score > 83). Tear film osmolarity was analyzed by the TearLab osmometer. Results: In our study, patients with moderate to severe keratoconjunctivitis sicca showed a tear film osmolarity of 320 mOsmol/L (301–324 mOsmol/L). The results of the control group were 301 mOsmol/L (298–304 mOsmol/L). Our results revealed a significantly higher tear film osmolarity in patients with moderate to severe keratoconjunctivitis sicca compared with the control group. The sensitivity was 87%, and the specificity was 81%. Conclusions: Our results approved the referent value in moderate to severe dry eye of approximately 316 mOsmol/L, as described in the literature. The results showed a significantly higher tear film osmolarity in patients with severe keratoconjunctivitis sicca compared with the healthy controls. Testing tear film osmolarity can be a very effective objective diagnostic tool in the diagnosis of dry eye disease.

Das trockene Auge

ZusammenfassungDie Pathogenese des trockenen Auges ist multifaktoriell und komplex. Neue Forschungsergebnisse zeigen, dass sich ein Entzündungsprozess der Augenoberfläche als „gemeinsame Endstrecke“ aller chronischen Formen des trockenen Auges herauskristallisiert. Daneben sind lokale Hormonimbalanzen, vor allem ein Androgenmangel, entscheidend für dessen Entstehung. Aktuelle Konzepte zur Pathogenese des trockenen Auges und die diagnostischen Basis- und Zusatzverfahren werden erläutert. Die In-vivo-Konfokalmikroskopie erlaubt erstmals die Quantifizierung von Entzündungszellen der Augenoberfläche zur Indikationsstellung und Verlaufskontrolle antientzündlicher Therapieoptionen des trockenen Auges.AbstractThe etiology of dysfunctional tear syndrome (“dry eye”) is multifactorial and complex. Recent evidence suggests an important role of androgens in regulating tear film secretion onto the ocular surface. In addition, inflammatory processes of the ocular surface seem to be the common final pathway of all chronic forms of dry eye. Novel concepts of pathogenesis and state-of-the-art diagnostic tools are discussed. In vivo confocal microscopy allows quantification of ocular surface inflammatory cells. This is of increasing importance for evaluation of anti-inflammatory treatments in dry eye patients.

[The dry eye. Current concepts on classification, diagnostics, and pathogenesis].

ZusammenfassungDie Pathogenese des trockenen Auges ist multifaktoriell und komplex. Neue Forschungsergebnisse zeigen, dass sich ein Entzündungsprozess der Augenoberfläche als „gemeinsame Endstrecke“ aller chronischen Formen des trockenen Auges herauskristallisiert. Daneben sind lokale Hormonimbalanzen, vor allem ein Androgenmangel, entscheidend für dessen Entstehung. Aktuelle Konzepte zur Pathogenese des trockenen Auges und die diagnostischen Basis- und Zusatzverfahren werden erläutert. Die In-vivo-Konfokalmikroskopie erlaubt erstmals die Quantifizierung von Entzündungszellen der Augenoberfläche zur Indikationsstellung und Verlaufskontrolle antientzündlicher Therapieoptionen des trockenen Auges.AbstractThe etiology of dysfunctional tear syndrome (“dry eye”) is multifactorial and complex. Recent evidence suggests an important role of androgens in regulating tear film secretion onto the ocular surface. In addition, inflammatory processes of the ocular surface seem to be the common final pathway of all chronic forms of dry eye. Novel concepts of pathogenesis and state-of-the-art diagnostic tools are discussed. In vivo confocal microscopy allows quantification of ocular surface inflammatory cells. This is of increasing importance for evaluation of anti-inflammatory treatments in dry eye patients.

Prospective, Randomized, Controlled Comparison of SYSTANE UD Eye Drops Versus VISINE INTENSIV 1% EDO Eye Drops for the Treatment of Moderate Dry Eye

PURPOSE The aim of this prospective, randomized, clinical, single-center study was to compare the safety and efficacy of 2 ocular surface lubricant eye drops: preservative-free hydroxypropyl (HP)-Guar (SYSTANE UD(®)) eye drops versus preservative-free Tamarindus indica seed polysaccharide (TSP) 1% (VISINE INTENSIV 1% EDO(®)) eye drops. METHODS Fifty-six eyes of 28 patients with moderate keratoconjunctivitis sicca (DEWS severity level 2) were enrolled in the trial. Patients were randomized for 2 treatment groups (SYSTANE UD eye drops vs. VISINE INTENSIV 1% EDO eye drops). The eye drops in both groups were applied 5 times per day for 3 months. Statistical analyses were performed using Statistica™ software (Mann-Whitney U-test and Wilcoxon test). P-Values<0.05 were considered significant. RESULTS After 3 months of treatment the patients of both groups had subjective benefit in the relief of symptoms of dry eye disease evaluated by the Ocular Surface Disease Index (OSDI) questionnaire score. Patients treated with HP-Guar and TSP showed improvements in tear film stability measured by tear break-up time (TBUT), which are statistically significant in the HP-Guar group (P=0.02). CONCLUSIONS The results of this clinical trial show improvements of symptoms and signs in patients with moderate dry eye after the consistent use of preservative-free HP-Guar and TSP lubricant eye drops. Both artificial tear formulations produce amelioration in tear film stability improving eye conditions and patient quality of life. HP-Guar seems to be slightly more effective in improving ocular surface protection by decreasing tear film evaporation.

Herstellung von Eigenserumaugentropfen zur ambulanten Therapie

BACKGROUND Autologous serum eye drops are an important therapy option in severe ocular surface disorders and the therapeutic effectiveness has been demonstrated in many clinical studies. The production and use of autologous serum eye drops is strictly controlled by legal regulations in Germany: Both the German Medicines Act (AMG) and the Blood Transfusion Act regulate production, distribution and application, unless it is carried out by one person under controlled conditions in a hospital setting. MATERIAL AND METHODS In cooperation with the ophthalmic clinic and the department of transfusion medicine, a standard operating procedure (SOP) was developed and a license for production and delivery of autologous serum eye drops was obtained from the appropriate local authorities. The experiences of the first two years of practice were analyzed. RESULTS By an interfaculty cooperation, the possibility of legal and feasible out-patient treatment with autologous eye drops has been established at the University Hospital Erlangen. From 07/2005 to 07/2007, there ware 240 prescriptions for autologous serum eye drops. Unexpectedly, a relatively high rate (3.3%) of patients with primarily unknown viral or bacterial infectious diseases were found, which were diagnosed during the screening. These patients had to be excluded from autologous serum eye drop therapy. CONCLUSION The treatment with autologous serum eye drops in an out-patient setting is possible, when the infrastructure for manufacture and delivery is provided in accordance with existing regulations.

[Manufacture of autologous serum eye drops for out-patient therapy : cooperation between ophthalmic clinic and transfusion medicine department].

BACKGROUND Autologous serum eye drops are an important therapy option in severe ocular surface disorders and the therapeutic effectiveness has been demonstrated in many clinical studies. The production and use of autologous serum eye drops is strictly controlled by legal regulations in Germany: Both the German Medicines Act (AMG) and the Blood Transfusion Act regulate production, distribution and application, unless it is carried out by one person under controlled conditions in a hospital setting. MATERIAL AND METHODS In cooperation with the ophthalmic clinic and the department of transfusion medicine, a standard operating procedure (SOP) was developed and a license for production and delivery of autologous serum eye drops was obtained from the appropriate local authorities. The experiences of the first two years of practice were analyzed. RESULTS By an interfaculty cooperation, the possibility of legal and feasible out-patient treatment with autologous eye drops has been established at the University Hospital Erlangen. From 07/2005 to 07/2007, there ware 240 prescriptions for autologous serum eye drops. Unexpectedly, a relatively high rate (3.3%) of patients with primarily unknown viral or bacterial infectious diseases were found, which were diagnosed during the screening. These patients had to be excluded from autologous serum eye drop therapy. CONCLUSION The treatment with autologous serum eye drops in an out-patient setting is possible, when the infrastructure for manufacture and delivery is provided in accordance with existing regulations.

Palate Lung Nasal Clone (PLUNC), a Novel Protein of the Tear Film: Three-Dimensional Structure, Immune Activation, and Involvement in Dry Eye Disease (DED).

PURPOSE Palate Lung Nasal Clone (PLUNC) is a hydrophobic protein belonging to the family of surfactant proteins that is involved in fluid balance regulation of the lung. Moreover, it is known to directly act against gram-negative bacteria. The purpose of this study was to investigate the possible expression and antimicrobial role of PLUNC at the healthy ocular surface and in tears of patients suffering from dry eye disease (DED). METHODS Bioinformatics and biochemical and immunologic methods were combined to elucidate the structure and function of PLUNC at the ocular surface. Tissue-specific localization was performed by using immunohistochemistry. The PLUNC levels in tear samples from non-Sjögrens DED patients with moderate dry eye suffering either from hyperevaporation or tear deficiency were analyzed by ELISA and compared with tears from healthy volunteers. RESULTS Palate Lung Nasal Clone is expressed under healthy conditions at the ocular surface and secreted into the tear film. Protein modeling studies and molecular dynamics simulations performed indicated surface activity of PLUNC. In vitro experiments revealed that proinflammatory cytokines and bacterial supernatants have only a slight effect on the expression of PLUNC in HCE and HCjE cell lines. In tears from DED patients, the PLUNC concentration is significantly increased (7-fold in evaporative dry eye tears and 17-fold in tears from patients with tear deficiency) compared with healthy subjects. CONCLUSIONS The results show that PLUNC is a protein of the tear film and suggest that it plays a role in fluid balance and surface tension regulation at the ocular surface.

Ophthalmologische Komplikationen beim Sjögren-Syndrom

ZusammenfassungHintergrundOkuläre Komplikationen sind eine typische Folge des primären und sekundären Sjögren-Syndroms. Da diese bei unzureichenden Therapiemaßnahmen bis zur Erblindung führen können, sollen diagnostische Verfahren und Therapiemöglichkeiten dargestellt werden.MethodenLiteraturübersicht aus PubMed und eigene klinische und experimentelle Daten.ErgebnisseBeim primären und sekundären Sjögren-Syndrom kommt es zu zahlreichen Komplikationen am Auge. Im Vordergrund stehen Störungen des präkornealen Tränenfilms.SchlussfolgerungenEine frühzeitige und enge Kooperation zwischen Rheumatologen und Augenarzt kann das Entstehen von visusbedrohenden Langzeitkomplikationen beim Sjögren-Syndrom vermeiden.AbstractBackgroundOcular complications are typical sequels of primary and secondary Sjögren’s syndrome. Since these can lead to blindness in the case of insufficient therapeutic steps, diagnostic tools and therapeutic options in Sjögren-syndrome associated ocular diseases are outlined.MethodsLiterature review from PubMed and own clinical and experimental results.ResultsNumerous ocular complications can occur both in primary and secondary Sjögren’s syndrome. The main problems involve tear film disturbances leading to severe forms of dry eye.ConclusionEarly and close cooperation between rheumatologist and ophthalmologist can avoid long-term complications in patients with Sjögren’s disease and ocular complications.

Diagnosis of X-Linked Hypohidrotic Ectodermal Dysplasia by Meibography and Infrared Thermography of the Eye.

Abstract Purpose: X-linked hypohidrotic ectodermal dysplasia (XLHED) is the most common form of ectodermal dysplasia. Clinical characteristics include meibomian gland disorder and the resulting hyperevaporative dry eye. In this study, we evaluated meibography and ocular infrared thermography as novel methods to diagnose XLHED. Methods: Eight infants, 12 boys and 14 male adults with XLHED and 12 healthy control subjects were subjected to a panel of tests including the ocular surface disease index (OSDI), meibography and infrared thermography, non-invasive measurement of tear film break-up time (NIBUT) and osmolarity, Schirmer’s test, lissamine green staining and fluorescein staining. Sensitivity and specificity were determined for single tests and selected test combinations. Results: Meibography had 100% sensitivity and specificity for identifying XLHED. Infrared thermography, a completely non-invasive procedure, revealed a typical pattern for male subjects with XLHED. It was, however, less sensitive (86% for adults and 67% for children) than meibography or a combination of established routine tests. In adults, OSDI and NIBUT were the best single routine tests (sensitivity of 86% and 71%, respectively), whereas increased tear osmolarity appeared as a rather unspecific ophthalmic symptom. In children, NIBUT was the most convincing routine test (sensitivity of 91%). Conclusions: Meibography is the most reliable ophthalmic examination to establish a clinical diagnosis in individuals with suspected hypohidrotic ectodermal dysplasia, even before genetic test results are available. Tear film tests and ocular surface staining are less sensitive in children, but very helpful for estimating the severity of ocular surface disease in individuals with known XLHED.