M. Kralinger

Ocular Delivery of Acetylsalicylic Acid by Repetitive Coulomb-Controlled Iontophoresis

To investigate the potential of transscleral coulomb-controlled iontophoresis (CCI) for repetitive delivery of acetylsalicylic acid (ASA) into the eye, a total of 50 rabbits was included in this study. Fourteen animals received serial CCI treatment. Fourteen animals underwent CCI with either ASA or balanced salt solution (BSS) for at least 6 days at 24- and 48-hour intervals. Eighteen animals received a single CCI application, while 18 animals were injected with 15 mg ASA/kg body weight intravenously. HPLC analysis was performed to determine the levels of salicylic acid (SA) in ocular tissues. Apart from clinical follow-up, 2 rabbits in the ASA and BSS groups were examined by electroretinography, and 2 animals were examined histologically. Though high concentrations of SA were measured, no alterations were observed clinically, histologically and electrophysiologically. Repetitive CCI demonstrated its potential as a topical drug delivery system for ASA into the eye. This transscleral delivery of ASA resulted in significant and sustained intraocular concentrations of SA without side effects. Iontophoresis may be advantageous in clinical administration maintaining therapeutic levels of ASA while avoiding adverse effects associated with the systemic administration of nonsteroidal anti-inflammatory drugs.

Retinal Thickness Analysis in Subjects with Different Refractive Conditions

Purpose: The aim of the present study is to evaluate a measure of retinal thinning at the posterior pole in eyes with increasing axial myopia. Methods: 129 eyes of 79 healthy persons with different refractive conditions were examined using a commercially available prototype of retinal thickness analyzer. Results: Increasing myopia has been found to be accompanied by bulbus elongation, and calculation of the values obtained with the instrument revealed a significant decrease in retinal thickness at the posterior pole in eyes with increasing axial myopia, i.e. 4.8 μm/spherical equivalent in the foveolar region and 6.6 μm/spherical equivalent in the other areas of the posterior pole. Conclusion: Our data show retinal thinning at the posterior pole in myopic eyes based for the very first time on in vivo measurements. Furthermore, our findings might influence measurement data for other purposes, if the degree of myopia is not taken into account.

The effect of streptozotocin-induced diabetes mellitus on substance P and calcitonin gene-related peptide expression in the rat trigeminal ganglion

Substance P (SP) and calcitonin gene-related peptide (CGRP) constitute the main sensory peptides in the trigeminal ganglion (TG). The objective of this study was to characterize peptidergic changes in the streptozotocin-induced diabetes mellitus rat model both quantitatively and qualitatively. Diabetes mellitus was induced by a single intraperitoneal injection of streptozotocin (65 mg/kg) and the levels of SP and CGRP were measured by means of radioimmunoassay (RIA) in a time-dependent manner. Peptide immunoreactivities were characterized by high pressure liquid chromatography (HPLC). The expression of both neuropeptides was examined 5 weeks after streptozotocin injection using in situ hybridization with 35S-labelled oligonucleotides. Saline-injected rats served as controls. SP was significantly decreased in the diabetic rat TG, i.e. , a 44.6% (+/-10.9) decrease after 1 week, 40.2% (+/-11.8) after 3 weeks and 72.3% (+/-14.6) after 5 weeks. CGRP was decreased only after 5 weeks (19.6% decrease +/-3.9), whereas at later stages, both peptide levels returned to normal values. HPLC revealed one major peak coeluting with the synthetic peptides. By using in situ hybridization, a significantly increased signal of both peptide-encoding mRNAs was found (43.8%), which seems to act to restore a diabetes-associated depletion of neuropeptides in the diabetic rat TG. The decreased SP- and CGRP levels in the diabetic rat TG reflect a diabetes-associated deficit which may be clinically relevant. Diabetes mellitus is associated with a variety of ocular complications, even corneal complications, including decreased corneal sensitivity, which in many ways resemble those after interruption of the normal trophic innervation of the eye. Our results point to reduced availability of neuropeptides for corneal innervation and may thus support the idea of a partial loss of trophic influences from the trigeminal nerve in diabetics.

Slow release of acetylsalicylic acid by intravitreal silicone oil

Purpose To assess in vitro the potential of silicone oil as a delivery system for acetylsalicylic acid (ASA) and to evaluate in vivo the pharmacokinetic distribution of salicylic acid (SA) in the eye. Methods In an experimental model ASA/silicone oil suspension mixed to a concentration of 1.67 mg/mL was investigated for release rate of ASA and SA. In vivo vitrectomy and intravitreal injection of two different ASA/silicone oil suspensions, both mixed to a concentration of 1.67 mg/mL, was performed on two groups, A and B, of New Zealand white rabbits. Salicylic acid concentrations in ocular tissues, aqueous, vitreous, and blood plasma were evaluated at 6 hours, 24 hours, and 5 days using high performance liquid chromatography. Results Salicylic acid was detected in all tissues. The highest levels were obtained in the vitreous: 745.4 &mgr;g/mL (A) and 640.0 &mgr;g/mL (B) at 6 hours. The retina followed with 332.9 ng/mg (A) and 281.3 ng/mg (B) at 6 hours and 31.6 ng/mg (A) and 48.1 ng/mg (B) at day 5. The maximum blood plasma levels were 5.2 &mgr;g/mL. Conclusion Silicone oil is an efficacious delivery system of ASA in vitro and in vivo. Higher concentrations of SA were found in all ocular tissues and fluids when compared to intravenous administration of maximum doses.

Elevated levels of calcitonin gene-related peptide in aqueous humor of patients with proliferative vitreoretinopathy.

Abstractu2002Purpose: To detect the levels of the sensory peptide calcitonin gene-related peptide in aqueous humor of patients with proliferative vitreoretinopathy and to compare them with those of uninflamed eyes (cataract and uncomplicated rhegmatogenous retinal detachment). Materials and methods: Using a highly specific and sensitive radioimmunoassay the concentration of calcitonin gene-related peptide was detected in fresh samples of aqueous humor obtained via paracentesis. Furthermore, calcitonin gene-related peptide-like immunoreactivities were characterized by high-pressure liquid chromatography.u2002Results: The mean level of calcitonin gene-related peptide was 6.11 fmol/ml in cataract controls and 14.77 fmol/ml in uncomplicated rhegmatogenous retinal detachment. In the cataract group, 9 of 18 cases were below the detection limit and in the retinal detachment group, 5 of 16. In proliferative vitreoretinopathy, the peptide averaged 76.92 fmol/ml and none of the samples was below the detection limit. High-pressure liquid chromatography revealed one major peak corresponding to synthetic calcitonin gene-related peptide. Conclusion: In recent studies, we found elevated levels of the sensory peptide substance P in aqueous humor of patients with proliferative vitreoretinopathy. This fact and the present result, the elevation of calcitonin gene-related peptide in aqueous humor of eyes with proliferative vitreoretinopathy, clearly point to an involvement of sensory peptides in the pathobiology of the disease. The source of the elevation is not clear, but we hypothesize that it originates from a neurogenic mechanism, i.e. an acceleration of the peptides by their enhanced release from the iris/ciliary body complex subsequent to sensitization of sensory neurons, thus representing a very interesting epiphenomenon of proliferative vitreoretinopathy. Our results constitute novel aspects in the pathophysiological concept of proliferative vitreoretinopathy and extend the knowledge about the pathobiology of the disease process.

Elevated levels of secretoneurin in the rabbit aqueous humor in response to formaldehyde irritation

BackgroundSecretoneurin, a 33-amino-acid neuropeptide, is generated by proteolytic processing of secretogranin II, which belongs to the chromogranin family. This study aimed to investigate whether secretoneurin is present in the uninflamed rabbit aqueous humor and whether it is released in response to treatment with topical formaldehyde, an agent known to release sensory peptides originating from the trigeminal ganglion.MethodsBlood samples and aqueous humor of eyes pretreated with neutral formaldehyde and untreated controls were analyzed for secretoneurin immunoreactivity by a highly sensitive radioimmunoassay. Furthermore, the molecular form of the secretoneurin immunoreactivity was characterized by gel filtration high-performance liquid chromatography (HPLC).ResultsIn the blood, secretoneurin levels were found to be below the detection limit of 2xa0fmol/100xa0μl. In the aqueous humor, secretoneurin-immunoreactivity was detected in moderate but significant amounts. The mean concentration of secretoneurin was 8.1 (±0.34) fmol/100xa0μl in controls and 7.8 (±0.1) fmol/100xa0μl 15xa0min after formaldehyde application. Thirty minutes after treatment, the secretoneurin levels were significantly elevated by 63%. Gel filtration HPLC revealed that the main molecular form corresponded to the free peptide secretoneurin.ConclusionsThe neuropeptide secretoneurin has been detected in the anterior segment of the eye for the first time. The elevation of secretoneurin in formaldehyde-treated eyes may be induced by an enhanced release from the iris/ciliary body complex, as formaldehyde is known to provoke neurogenic inflammation in the anterior segment via release of sensory peptides originating from the trigeminal ganglion. This is why our results indicate a sensory origin of secretoneurin in the eye.

Long-term visual results after vitrectomy in diabetic macular edema

Since the introduction of lasercoagulation, therapy of diabetic macular edema, which is the most frequent cause for loss of vision in diabetic patients, had become possible. The accumulation of intraretinal fluid due to hyperpermeability of the retinal vasculature may lead to focal, diffuse and cystoid edema [1].