Biomedical subjects

G Thumann

Publications and source records attributed to G Thumann.

29 records · Page 2Linked to original sources

PubMed1998

The mRNA expression of cytokines and their receptors in cultured iris pigment epithelial cells: a comparison with retinal pigment epithelial cells.

It has been suggested that human iris pigment epithelial (IPE) cells isolated from iridectomized tissue could be used as autologous cells for transplantation into the subretinal space in diseases with dysfunctional retinal pigment epithelium (RPE). RPE cells synthesize a number of cytokines and their receptors which are important for its proper function. Nearly nothing is known about the capacity of IPE to synthesize cytokines or responding to them. To compare the mRNA expression of 36 cytokines or their receptors in cultured adult IPE cells and RPE cells we used semi-quantitative reverse transcription polymerase chain reactions (RT-PCR). Included in our assay were cytokines with known expression in RPE to get a broad basis for comparing IPE cells: basic fibroblast growth factor (bFGF or FGF-2), and one of its receptor (FGFR-1), epidermal growth factor (EGF), and its receptor EGF-R, transforming growth factor beta(TGFbeta), and its type III receptor TGFbeta-R3, the platelet-derived growth factors and receptors (PDGF A, PDGF B, PDGF-Ralpha, PDGF-Rbeta), tumor necrosis factor alpha(TNFalpha), and two receptors TNF-R1 and TNF-R2, insulin (INS) with receptor INS-R, insulin-like growth factors (IGF1, IGF2), and receptors (IGF1-R, IGF2-R), vascular endothelial growth factor (VEGF), and two receptors (VEGF-R1 or FLT-1 and VEGF-R2 or FLK-1), the receptor for VEGF-C: VEGF-R3 or FLK-4, interleukin 6 (IL6), and its receptor (IL6-R), nerve growth factor (NGF), interleukin 1alpha(IL1alpha), and a receptor (IL1-R). In addition, cytokines or their receptors not known to be expressed in RPE were included to widen our picture of cytokine gene expression in the eye: stem cell factor (SCF), its receptor (SCF-R), low-affinity nerve growth factor receptor p75 (p75(NGF-R), ciliary neutrothropic factor (CNTF), and its receptor (CNTF-R), glycoprotein 130 interleukin 6 transducer (gp130 (IL6-SD), leukemia inhibitory factor (LIF), and its receptor (LIF-R). Semi-quantitative expression data were obtained using series of fivefold dilutions of each cDNA and a fixed number of PCR cycles. The expression of RPE 65, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and beta2-microglobulin (B2MG) was used as a control for cellular origin, RNA quality and PCR conditions. With the exception of insulin and tumor necrosis factor alphaall other cytokines analysed and their receptors were expressed in both IPE and RPE cells, even though the levels varied. No qualitative or quantitative difference were observed in the mRNA expression level of 34 (94%) of the cytokines or receptors between IPE and RPE. In contrast, the mRNA expression level of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor 2 [VEGF-RS (FLK-1)] was lower in IPE than in RPE cells. As an increased expression of VEGF in the RPE in maculae with age-related macular disease could be involved in its pathogenesis, a decreased expression of angiogenic growth factors in IPE cells could possibly be beneficial for the therapy of age-related maculopathy if indeed other tasks of non-functional RPE cells could be performed by IPE cells. The similarity of the mRNA expression pattern in 94% of the cytokines analyzed supports the assumption that IPE cells potentially can perform functions of RPE cells in the appropriate environment.

Adult↗

PubMed1998

Phagocytosis of rod outer segments by human iris pigment epithelial cells in vitro.

BACKGROUND: We set out to evaluate the growth potential of human iris pigment epithelial (hIPE) cells in vitro, to establish whether these cells acquire the ability to phagocytose rod outer segments (ROS) and to compare the phagocytic activity of hIPE to that of human retinal pigment epithelial (hRPE) cells. METHODS: hIPE and hRPE cells were isolated and cultured from human donor eyes and surgical specimens and growth characteristics were analyzed. HIPE and hRPE of an eye of a 46-year-old donor were used for the phagocytosis assay. Phagocytosis was evaluated by adding ROS isolated from porcine retina to cultures of hIPE and hRPE, which had been labeled with the pH-sensitive fluorescent dye, carboxy-SNAFL. After 4 h the number of ingested ROS was counted with a light microscope. For each cell type phagosomes in 500 cells were counted. The epithelial characteristics of the cells used in this study were evidenced by their morphology. RESULTS: Morphologically cultured hIPE are indistinguishable from the hRPE cultured from the same donor eye and show a similar pattern of cytokeratin distribution. Cultured hIPE acquire the ability to phagocytose ROS at a level slightly lower than hRPE; hIPE contained 0.76 phagosomes per cell, hRPE 0.99 phagosomes per cell. CONCLUSION: The morphology of hIPE in culture and the acquisition of the phagocytic phenotype indicate that these cells have the ability to differentiate into cells that have characteristics in common with hRPE. The acquisition of phagocytic activity suggests that it is feasible to culture hIPE from surgical iridectomies and that these cultured cells can be transplanted into the subretinal space in individuals with retinal degenerations.

Cells, Cultured↗

PubMed1997

Apoptosis in proliferative vitreoretinal disorders: possible involvement of TGF-beta-induced RPE cell apoptosis.

The targeted induction of apoptosis is a novel therapeutic approach to control the unlimited growth of proliferating cells. Since massive proliferation of cells at the vitreoretinal interface is a key feature of proliferative vitreoretinal disorders, we sought to identify apoptosis in epiretinal membranes from patients with proliferative vitreoretinopathy (PVR), proliferative diabetic retinopathy (PDR) and macular pucker. Further, we evaluated the possible induction of apoptosis of retinal pigment epithelial (RPE) cells by transforming growth factor-beta(TGF-beta). Apoptotic cells were identified by in situ DNA end labeling and acridine orange staining on paraffin-embedded tissue sections from epiretinal membranes of patients with all vitreoretinal disorders examined. Labeled nuclei or condensed chromatin were scattered throughout the membranes or occurred in clusters. Most apoptotic cells were RPE-derived, as assessed by cytokeratin immunochemistry. No apoptotic glial cells were detected. In PVR, proliferative activity, as confirmed by Ki-67 immunochemistry, was associated with short history and rapid disease progression. Apoptotic nuclei were observed more frequently in long-standing PVR or slow progression towards traction retinal detachment. TGF-beta was detected in all control vitreous samples by bioassay at concentrations below 20 ng ml-1. TGF-beta levels increased up to 20-fold in pathological vitreous. Marked heterogeneity was observed in all patient groups. The degree of TGF-beta activation was significantly higher in PVR than in PDR. Proapoptotic effects of TGF-beta were demonstrated in cultured human RPE cells by electron microscopy, in situ DNA end labeling, comet assay and a photometric enzyme immunoassay for histone-associated DNA fragments. Apoptosis appears to be a key regulatory mechanism of growth control of specific cell populations in proliferative vitreoretinal disorders. Administration of proapoptotic growth factors such as TGF-beta may provide a novel approach to inhibit cellular proliferation at the vitreoretinal interface.

Apoptosis↗

PubMed1997

Branch retinal artery occlusion by diathermy of a persistent hyaloid artery.

PURPOSE: To report branch retinal artery occlusion caused by diathermy of a persistent hyaloid artery. METHODS: Case report. A 21-year-old woman with unilateral tractional retinal detachment caused by contraction of the persistent primary vitreous underwent vitrectomy with endocoagulation of the primary hyaloid artery. RESULTS: Postoperatively, the patient had opacification of the neurosensory retina in the area of the inferotemporal branch retinal artery accompanied by a corresponding visual field defect. CONCLUSIONS: Diathermy of a persistent hyaloid artery may be complicated by the occlusion of a branch retinal artery. In similar cases, atypical branches of retinal arteries must be considered. Dissection of the anomaly from the posterior lens capsule should be performed without coagulation or perhaps endoscopically if visualization is obscured by a Mittendorf dot.

Adult↗

PubMed1997

[Vitrectomy in treatment of eyes with complicated uveitis].

BACKGROUND: Vitrectomy serves as an important treatment modality in uveitis, removing inflamed vitreous body and eliminating secondary complications. The aim of our retrospective study was to evaluate the value of vitrectomy in the treatment of complications in uveitis. PATIENTS AND METHOD: We included 50 eyes of 50 patients suffering from chronic uveitis, which underwent vitrectomy surgery during the period of 1990 to 1993. The study only included patients who had vitrectomy for uveitis for the first time. Indications leading to surgery were density of optic media, retinal detachment, cyclitic or preretinal membranes. The mean follow-up was 455 (range 48-1420) days. RESULTS: Vitrectomy was performed at a mean delay of 452 days after initial presentation. The mean visual acuity at the time of surgery was 0.05. The best visual acuity postoperatively was measured at a mean time of 118 days after vitrectomy. Overall, at the last control, eyes with preoperative retinal detachment presented a much worse final functional result (visual acuity of 0.02) than eyes without detachment (visual acuity of 0.5). Ten eyes resulted in hypotonia after surgery (7 eyes with persisting hypotonia). Most frequent complications in the postoperative course were macular pucker (11 eyes), secondary glaucoma (10 eyes), macular edema (6 eyes) and retinal detachment (6 eyes). Fifty percent of all complications occurred within the first 3 months following surgery. CONCLUSION: Vitrectomy leads to a permanent increase in visual acuity through the treatment of complications, on the condition that a stable retinal situation was present preoperatively.

Adult↗

PubMed1997

Descemet's membrane as membranous support in RPE/IPE transplantation.

PURPOSE: The correct orientation of retinal pigment epithelium (RPE) cells is necessary for the integrity and proper function of the retina. For transplantation of RPE/iris pigment epithelium (IPE) grafts to the subretinal space in age-related macular degeneration, this cellular orientation is most effectively provided by a membranous support. The goal of this study was to establish an autologous or homologous membrane as a substratum for the growth of RPE/IPE. METHODS: Porcine and bovine RPE and IPE were placed in primary culture on a dissected sheet (5 x 5 mm) of autologous porcine and bovine Descemet's membrane in slide chambers and grown to confluence. RESULTS: RPE and IPE cells cultured on Descemet's membrane form an intact monolayer. Light and electron microscopy showed the formation of both an intact monolayer and microvilli in both cell types. CONCLUSION: Since the slow host-graft rejection appears to play an important role in the failure of RPE transplantation in the subretinal space, it is critical to be able to transplant autologous materials. The techniques presented here establish a novel means to culture RPE or IPE cells on autologous Descemet's membrane where they form a "cell monolayer patch," consisting of a fragment of Descemet's membrane with cultured RPE or IPE, which can be easily manipulated and transplanted, using an established glass pipette method.

Animals↗

PubMed1997

The artificial iris diaphragm for vitreoretinal silicone oil surgery.

BACKGROUND: Silicone keratopathy frequently develops as a complication of silicone oil tamponade in the management of severe trauma in eyes with partial or complete aniridia. We therefore designed an "open" artificial iris diaphragm to prevent silicone oil-endothelial contact. In hypotony, where insufficient circulation of aqueous allows silicone in the anterior chamber despite an open diaphragm, a new solution became necessary. The "closed" artificial diaphragm was developed. METHODS: In this retrospective study, two consecutive series of artificial iris diaphragms were compared. Forty-four patients received either an open type (20 eyes) or a closed type (24 eyes) and were observed for 409 +/- 421 days (range, 32-1912). All eyes were aphakic, normotonous, and had a traumatic, compromised iris diaphragm or were aphakic and hypotonic as a result of injury, proliferative vitreoretinopathy, proliferative diabetic retinopathy, or uveitis, with an intact natural iris diaphragm. RESULTS: Silicone oil was retained behind the open diaphragm throughout the observation period in 40% of the eyes. Major long-term complications were hypotony and fibrous overgrowth. Silicone was retained behind the closed diaphragm in 50% of the eyes. CONCLUSION: Because proliferative vitreoretinopathy is active for months and multiple surgical interventions become necessary to avoid phthisis in eyes with highly pathologic changes, longstanding or permanent silicone tamponade is used. The artificial diaphragm prevents silicone-corneal contact in approximately 50% of aphakic eyes for at least 1 year.

Adolescent↗

PubMed1996

[Effect of the contour line on cup surface using the Heidelberg Retina Tomograph].

BACKGROUND: Significance of topometric follow-up examinations of the optic nerve head in glaucomatous eyes depends on the reproducibility of the calculated parameters. Since the definition of the standard reference plane in software version 1.11 of the Heidelberg Retina Tomograph has been changed, intrapapillary parameters depend directly on the position of the contourline in the sector between -10 degrees to -4 degrees, and therefore on the observer variability to determine the disc border. We evaluated intra- and interobserver variability and present a simple approach to increase reproducibility. METHOD: The disc border of 4 glaucomatous eyes, 3 ocular hypertensive eyes and 3 eyes of healthy subjects were traced by two observers, 5 times using the free draw mode and 5 times by the addition of contourline circles. RESULTS: We found a median variability of the mean disc radius in sector -10 degrees to -4 degrees of 51 microns, which defines the position of the standard reference plane, resulting in a median variability of the position of the standard reference plane of 33 microns which caused a variability of 81 microns2 of the cup area. Addition of contourline circles smoothing the final contourline along the border of the optic disc resulted in a decrease of the coefficient of variation of the standard reference plane of 3.76% (6.76% vs. 3.0%), of the cup area of 2.34% (3.87% vs. 1.53%) and of the rim volume of 3.41% (9.75% vs. 6.34%). CONCLUSION: The calculation of the cup area using software version 1.11 of the Heidelberg Retina Tomograph depends on observer variability. The addition of contourline circles to define the final contourline along the disc border increases reproducibility. However, in follow-up of topometric examinations of the optic nerve head the software supported transfer mode should be used. Comparing topometric data of an individual optic disc in follow-up suppose the same definition of the contourline. Therefore, topometric data evaluated using software version 1.10 or earlier needs to be recalculated.

Adolescent↗

PubMed1996

[Risks and therapy options in dural arteriovenous fistulas between branches of the external carotid artery and the cavernous sinus. A case report and review of the literature].

INTRODUCTION: Dural arteriovenous fistula of the Arteria carotis externa and the sinus cavernosus are rare vascular malformations. In the literature the prognosis and necessity of treatment is described very variable. PATIENTS AND METHODS: A 90 year old female presents in our outpatient clinic with acute exophthalmus, dilated episcleral veins, cerebral bruits and beginning orbital apex syndrome. After normal computerized axial tomography (CAT) without and with contrast medium, we were able to diagnose a dural AV-fistula by angiography. Shortly after dismission the patient had a spontaneous subarachnoidal haemorrhage with severe neurological symptoms. CONCLUSIONS: The suspect of an intracranial AV-malformation implies a detailed diagnostic search with precise localisation of the process. In any patient with intracerebral and intracranial fistula there has to be an individual check of possible and necessary treatment. It is impossible to classify dural AV-fistula as a benign disease with harmless clinical course.

Aged↗

PubMedDate not supplied

The normalised rim/disc area ratio line.

The assessment of the cup of the optic disc depends, among other criteria, on the disc area. A small cup in a small optic disc can indicate an advanced glaucomatous lesion, on the other hand a large cup in a large optic disc can be normal. Therefore, an individual normalised rim/disc area ratio line together with the curves of 50th percentile and the 95th percentile of normal could help to better distinguish between glaucomatous and normal optic cups. The aim of our study was to calculate and to evaluate such a normalised rim/disc area ratio line. Heidelberg Retina Tomograph examinations of the optic nerve head of 100 randomly selected eyes of 100 normal subjects were evaluated. We calculated the disc area adjusted rim/disc area ratio in sectors of 10 degrees. The 95th percentile and the 50th percentile of each of the 36 sectors were calculated. Based on these normal percentile lines it was possible to display an individual normalised rim/disc area ratio line in the topographic images of an individual optic disc examination. Here we demonstrated examples of a normal optic disc, optic nerve heads with moderate and advanced lesions and a small optic disc with glaucomatous damage. We present a new display mode of the results of Heidelberg Retina Tomograph optic nerve head examination, which may be helpful for an easy and reliable assessment of glaucomatous optic nerve head damage only looking at topographic images.

Adolescent↗

PubMedDate not supplied

Development and cellular functions of the iris pigment epithelium.

A number of studies have shown that transplantation of retinal pigment epithelial (RPE) cells to the subretinal space offers a promising treatment modality for retinal degenerative diseases. However, it is necessary to transplant autologous cells to avoid rejection; unfortunately, obtaining autologous RPE cells necessitates such traumatic surgical intervention as to make this approach irrelevant. It has been hypothesized that iris pigment epithelial (IPE) cells may be a possible substitute for RPE cells for transplantation into the subretinal space. The iris pigment epithelium, which has the same embryonic origin as retinal pigment epithelium, has not received much attention from visual scientists. Even though it forms a highly specialized tissue, it is not clear whether the iris pigment epithelium contributes critical functions to the health of the visual system. In vivo the IPE does not appear to have any of the functions characteristic of RPE; however, in vitro cultured IPE cells do acquire functions, such as specific phagocytosis of rod outer segments, that are characteristic of RPE cells, and have been shown to have the potential to carry out many functions characteristic of RPE cells, e.g., retinol metabolism. This review outlines the development and cellular functions of the IPE with special emphasis on the modulation of those functions that can allow the IPE cells to be transplanted to the subretinal space where they appear to acquire differentiated properties of retinal pigment epithelium (RPE).

Cell Transplantation↗