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Biomedical subjects

R Birngruber

Publications and source records attributed to R Birngruber.

At least 55 records · Page 3Linked to original sources

A comparison of retinal morphology viewed by optical coherence tomography and by light microscopy.

OBJECTIVE: To compare the cross-sectional images of primate retinal morphology obtained by optical coherence tomography (OCT) with light microscopy to determine the retinal components represented in OCT images. METHODS: Laser pulses were delivered to the retina to create small marker lesions in a Macaca mulatta. These lesions were used to align in vivo OCT scans and ex vivum histologic cross sections for image comparison. RESULTS: The OCT images demonstrated reproducible patterns of retinal morphology that corresponded to the location of retinal layers seen on light microscopic overlays. Layers of relative high reflectivity corresponded to horizontally aligned retinal components such as the nerve fiber layer and plexiform layers, as well as to the retinal pigment epithelium and choroid. In contrast, the nuclear layers and the photoreceptor inner and outer segments demonstrated relative low reflectivity by OCT. CONCLUSIONS: Retinal morphology and macular OCT imaging correlate well, with alignment of areas of high and low reflectivity to specific retinal and choroidal elements. Resolution of retinal structures by OCT depends on the contrast in relative reflectivity of adjacent structures. Use of this tool will enable expanded study of retinal morphology, both normal and pathologic, as it evolves in vivo.

Animals↗

[Comparative studies with various corneal topography measurement devices].

PURPOSE: It is difficult to measure the topography of the cornea with high resolution and visualize it on a map displaying refraction. This is demonstrated by the ongoing improvement and further development of different methods and by the fact that users of these techniques are not always satisfied. MATERIALS AND METHODS: Five different ring projectors were compared. Spherical and aspherical ball standards were used to measure the standard deviation of the refraction of the system. A patient group of eight people with healthy eyes was used to compare the measurement accuracy and operational errors. Some patients were also measured after cataract surgery. The lateral and axial range of the devices was determined. RESULTS: The measurement accuracy for ball standards for the five devices was below 1/8 D. For the in vivo case in the control group it was below 1/4 D for four devices. Furthermore, the reproducibility of the results, the accuracy of angle determination and the influence of mechanical and optical design will be discussed. The study demonstrates applications and limits of this measurement method.

Cataract Extraction↗

[Studies of the development of refractive effects in intrastromal refractive corneal surgery with the picosecond laser].

BACKGROUND: Picosecond laser intrastromal photorefractive keratectomy (ISPRK) aims at achieving a flattening of the central cornea by plasma-mediated tissue evaporation without affecting the anterior or posterior corneal layers. We investigated the laser-induced tissue effects to establish a functional relationship between laser parameters and tissue removal and to assess their influence on the healing process and long-term refractive changes. MATERIALS AND METHODS: A modified ISL 2001 System with a cone angle of 30 degrees was used for in vitro investigations of the laser effects in water and porcine cornea. Photographic methods were used to determine the plasma volume and the thickness of the laser-generated intrastromal bubble layer as a function of the pulse energy and the number and separation in which the pulses were applied (216 eyes). Histological evaluation was done by polarization microscopy (9 eyes). RESULTS: Polarization microscopy revealed only minor signs of thermal tissue damage. The maximum amount of tissue that can be evaporated without damaging the outer corneal layers corresponds to a layer about 10 microns thick. With a 6-mm optical zone, this tissue removal yields an immediate refractive effect of only 0.85 dpt. Stronger long-term refractive changes observed in animal experiments and clinical studies must thus be due to the healing response of the cornea. The healing response may be induced by mechanical distortion due to intrastromal bubble formation affecting about one third of the corneal thickness. CONCLUSION: Since the refractive effects are apparently strongly influenced by corneal healing, they are poorly predictable and can probably not be used for clinical purposes.

Animals↗

[Optical coherence tomography of the cornea and the anterior eye segment].

TARGET: The method of optical coherence tomography (OCT) was investigated regarding its suitability and limits for measuring the cornea and the anterior segment of the eye. Furthermore, the stromal expansion of thermally induced lesions in the cornea directly after irradiation was determined within the scope of the laser thermokeratoplasty (LTK). MATERIAL AND METHODS: With the experimental scanning OCT system, x-z sections of the anterior eye segment were made with an optical resolution of about 20 microns axially and 25 microns laterally. Freshly enucleated, tonicized porcine eyes were used as model eyes. Thermal lesions were applied with a continuously emitting laser diode (lambda = 1.86 microns) and various radiation parameters. Before and after coagulation, the cornea was viewed from limbus to limbus in a central OCT scan and the individual coagulation source was measured. RESULTS: Global and local changes of the thickness of the cornea as well as the distance between cornea and lens were measured with high precision. Thermal lesions in their expansion can be dearly presented and matching well with the histologically stained sections, but are not as exactly defined at the edges due to the limited optical resolution, as known from histological preparations. CONCLUSION: With the OCT method quantitative measuring of the anterior eye segment can be performed in vitro and with reduced resolutions also in vivo. Due to the qualitatively good correspondence regarding the dimensions of thermal damage of the cornea with histologically obtained morphometric results, this method can be used for supervision of coagulation directly after LTK as well as for examination of the individual healing process.

Animals↗

[Imaging of laser thermokeratoplasty lesions by optical low coherence tomography and polarization microscopy after Sirius Red staining].

BACKGROUND: Information on the extent and degree of the thermal effect produced is of great importance for control of the laser dosage in laser thermokeratoplasty (LTK) and for postoperative follow-up. We investigated on acute LTK effects which information images obtained by optical low coherence tomography (OCT) offer compared to those obtained by polarization microscopy. METHODS: Porcine eyes were irradiated through a 400 microns quartz fiber using light from a laser diode emitting up to 300 mW at a wavelength of 1.86 microns. Thermal lesions of varying strength were scanned using an experimental OCT device with about 25 microns lateral and 20 microns axial resolution. Histologic evaluation of the scanned areas was done by polarization microscopy after Sirlus-Red staining, and similar lesions were also analyzed by TEM. RESULTS: Both methods differentiated three damage zones a transition zone, a zone of moderate coagulation, and a central zone of strong coagulation. In the transition zone, increased birefringence was seen in polarization microscopy, which correlated with increased light scattering seen in the OCT images. In the moderately coagulated zone, a decrease in birefringence was associated with an even stronger increase of the OCT signal. In the central zone, a loss of the fibrillar tissue structure was observed, which led to a complete loss of birefringence and a strong reduction of the OCT signal. CONCLUSIONS: Although OCT does not provide the detailed information on thermal changes of tissue seen by the histologic method, it offers information on the extent and degree of tissue changes without preparation artifacts and provides a non-invasive method of immediate and follow-up control of LTK lesions. A quantitative analysis of changes in corneal thickness and curvature is much simpler than by a slit lamp. Time-resolved measurements of corneal light scattering may be used for on-line control of the laser-light dosage during LTK.

Animals↗

Argon laser retinal lesions evaluated in vivo by optical coherence tomography.

PURPOSE: To assess the in vivo evolution of argon laser retinal lesions by correlating the cross-sectional structure from sequential optical coherence tomography with histopathologic sectioning. METHODS: Argon laser lesions were created in the retinas of Macaca mulatta and evaluated by cross-section optical coherence tomography, which was compared at selected time points with corresponding histopathology. RESULTS: Argon laser lesions induced an optical coherence tomography pattern of early outer retinal relative high reflectivity with subsequent surrounding relative low reflectivity that correlated well with histopathologic findings. The in vivo optical coherence tomography images of macular laser lesions clearly demonstrated differences in pathologic response by retinal layer over time. CONCLUSION: The novel sequential imaging of rapidly evolving macular lesions with optical coherence tomography provides new insight into the patterns of acute tissue response by cross-sectional layer. This sequential imaging technique will aid in our understanding of the rapid evolution of retinal pathology and response to treatment in the research and clinical setting.

Animals↗

Influence of ablation plume dynamics on the formation of central islands in excimer laser photorefractive keratectomy.

PURPOSE: The aim of this study was to understand the dynamics of ablation products during excimer laser photorefractive keratectomy, and their influence on the formation of central islands. METHOD: Laser flash photography was used to investigate the dynamics of ablation products during photorefractive keratectomy. The ablation plume over polymethyl methacrylate (PMMA) and porcine cornea targets ablated with different zone diameters was imaged at various times between 10 musec and 100 msec after the ablating laser pulse. On PMMA targets, the profiles of the resulting ablation craters were measured. RESULTS: In all cases, the ablation products formed a ring vortex. The plume velocities on cornea were found to be approximately twice as fast as on PMMA for all zone diameters. For both materials, the ablation plume evolves faster for smaller zone diameters. Central islands were observed for zone diameters between 1 and 7 mm on PMMA substrates. The islands became more pronounced with increasing zone diameter. CONCLUSIONS: A major cause for the formation of central islands was found to be particle redeposition at the center of the ablation zone. Because of the vortex dynamics, redeposition of particles favorably occurs at the center of the ablation zone. Additionally, the dynamics of the ablation plume lead to a concentration of airborne particles over the center of the ablation zone, which also may contribute to the creation of central islands by partial absorption of the next excimer laser pulse.

Animals↗

Optical coherence tomography of the human skin.

BACKGROUND: Optical coherence tomography (OCT) is a new diagnostic method for tissue characterization. OBJECTIVE: We investigated normal and pathologic structures in human skin in several locations to evaluate the potential application of this technique to dermatology. METHODS: Based on the principle of low-coherence interferometry, cross-sectional images of the human skin can be obtained in vivo with a high spatial resolution of about 15 microns. Within a penetration depth of 0.5 to 1.5 mm, structures of the stratum corneum, the living epidermis, and the papillary dermis can be distinguished. RESULTS: Different layers could be detected that were differentiated by induction of experimental blisters and by comparison with corresponding histologic sections. Furthermore, OCT images of several skin diseases and tumors were obtained. CONCLUSION: OCT is a promising new imaging method for visualization of morphologic changes of superficial layers of the human skin. It may be useful for noninvasive diagnosis of bullous skin diseases, skin tumors, and in vivo investigation of pharmacologic effects.

Adult↗

Factors determining the refractive effects of intrastromal photorefractive keratectomy with the picosecond laser.

PURPOSE: To determine the relationship between laser parameters and tissue removal with picosecond laser intrastromal photorefractive keratectomy (ISPRK) and to assess the effect of the parameters on the healing process and the long-term refractive changes. SETTING: Medical Laser Center Lübeck, Germany. METHODS: A modified Intelligent Surgical Lasers (ISL) 2001 system with a cone angle of 30 degrees was used for in vitro investigations of the laser effects in water and on the porcine cornea. Photographic methods were used to determine the plasma volume and to investigate the thickness of the intrastromal bubble layer as a function of the laser pulse energy, the number of layers in which the pulses were applied, and the layer separation. The data were used to calculate the amount of tissue removal. Histologic evaluation was done by polarization microscopy after Sirius-red staining. RESULTS: The laser pulses produced a sponge-like appearance of the stroma in a layer extending through about one third the corneal thickness, accompanied by mechanical distortions of the stromal lamellae. Thermal changes were weak. Tissue removal was impaired by the cavitation bubbles from preceding laser pulses. The amount of evaporated tissue depended only weakly on laser pulse energy and number of layers in which the pulses were applied. The maximum amount of tissue that could be evaporated without damaging the outer corneal layers was a 10.0 microm thick layer. With a 6.0 mm optical zone, the tissue removal produced an immediate refractive effect of only 0.85 diopter. CONCLUSIONS: Intrastromal PRK does not work as originally envisioned because the amount of evaporated tissue is very small. The greater long-term refractive changes observed in animal experiments and clinical studies must therefore be due to the healing response of the cornea. Because the refractive effects are strongly influenced by the healing response of the cornea, they are poorly predictable.

Animals↗

Photodynamic targeting of human retinoblastoma cells using covalent low-density lipoprotein conjugates.

Combination of photosensitizers with carrier molecules has been shown to enhance the efficiency of photodynamic therapy (PDT). Owing to an increased expression of their receptors on some malignant and proliferating cells, low-density lipoproteins (LDLs) are potential endogenous carriers. A photosensitizer, chlorin e6 (Ce6), was covalently bound to LDL via carbodiimide activation. The Ce6-LDL conjugate was evaluated on a fibroblast cell line with defined LDL receptor expression and a retinoblastoma cell line (Y79). Uptake of free Ce6 and Ce6 either covalently bound to or complexed with LDL was measured by spectrofluorimetry. Phototoxicity after irradiation at 660 nm was determined by a mitochondrial activity assay (MTT). Covalent binding to LDL significantly increased the uptake of Ce6 for both cell lines by a factor of 4-5. A Ce6: LDL binding ratio of 50:1 was optimal. A receptor-mediated uptake was demonstrated by saturability and competitive inhibition by free LDL. Binding also occurred at 2 degrees C and was attributed to non-specific associations. Irradiation with 10 J cm-2 of 660 nm light after treatment of cells with Ce6-LDL conjugate reduced the MTT activity by 80%, while free or mixed Ce6 induced a maximum of 10% reduction in the MTT activity following identical treatment conditions. These data suggest that targeting of LDL receptor-bearing cells using covalently bound carriers, such as LDL, might increase the efficiency and selectivity of PDT. Intraocular tumours such as retinoblastomas could be appropriate targets for such an approach owing to the ease of access of light sources and the need for non-invasive approaches in sensitive ocular sites.

Carrier Proteins↗

Mitomycin-C in laser sclerostomy: benefit and complications.

BACKGROUND AND OBJECTIVE: The authors studied the effect of topical mitomycin-C administration on the maintenance of filter function and intraocular pressure (IOP) following laser sclerostomy. PATIENTS AND METHODS: Twenty-six patients with a presumed high risk of episcleral scarring were treated intraoperatively with topical mitomycin-C (0.5 mg/ml) for 3 to 5 minutes. Their IOPs were monitored for at least 2 years. The IOP data of these patients were compared with the results for 46 patients who underwent the identical procedure without antimetabolite. RESULTS: IOPs below 23 mm Hg were achieved in 70% of the mitomycin-C-treated patients. Compared with the non-mitomycin-C group, the rate and duration of early postoperative hypotony was significantly increased in the mitomycin-C-treated group. CONCLUSION: Mitomycin-C is useful for maintaining successful filter function in patients with unfavorable prognoses. However, severe and persistent hypotony may occur.

Administration, Topical↗

Ablation dynamics in laser sclerostomy Ab externo by means of pulsed lasers in the mid-infrared spectral range.

BACKGROUND AND OBJECTIVE: Sclerostomy ab externo with pulsed laser systems is currently in phase II clinical trials. The authors investigated the ablation dynamics of tissue treated with pulsed laser systems in the mid-infrared range to estimate the extent of thermo-mechanical damage to the sclera and the anterior chamber. MATERIALS AND METHODS: Freshly harvested porcine eyes were used. A bare 400-micron fiber in direct contact with tissue was used for fistulization. Polarization light microscopy, fast-flash photography, as well as optical and acoustic transients were performed for analysis. RESULTS: Substantial mechanical tissue deformation and dissections were found during pulsed laser ablation. The mechanical damage range within tissue far exceeds the pure thermal damage zone. Aspheric cavitation bubbles of up to 3 mm in length penetrate the anterior chamber after perforation. The cavitation demonstrates a significantly larger time constant in tissue than in water. CONCLUSIONS: Early fistula occlusions due to iris adherences may be attributed to iris trauma caused by cavitation. In response to the findings of this study, the authors propose an automatic feedback system to control the ablation process and minimize secondary ocular tissue effects. With respect to the overall damage zones, a new continuous-wave, mid-infrared diode laser system seems to be superior to pulsed laser systems.

Animals↗

Benzoporphyrin-lipoprotein-mediated photodestruction of intraocular tumors.

Benzoporphyrin derivative (BPD), a sensitizer currently in clinical trials, was evaluated for the treatment of experimental Greene melanoma implanted in the rabbit iris. To improve tumor targeting, BPD was complexed with low-density lipoprotein (LDL) representing an endogenous carrier system for BPD as previously described. Twelve tumors were irradiated at a sensitizer dose of 2 mg kg-1 body weight using a dye laser at 692 nm. Tumor responses were documented by photography, angiography and light and electron microscopy. All tumors treated with 80 J cm-2 regressed irreversibly. The principal mechanism of tumor necrosis was thrombosis following disruption of endothelial membranes. Ultrastructure data suggested tumor cell damage, although evidence for this being the result of direct PDT-mediated tumor cell death was less clear. These data suggest that BPD-LDL may be used to improve the selectivity of photodynamic tumor therapy possibly by the increased uptake of lipoprotein-delivered sensitizer to neovascular endothelial cells.

Animals↗

[Cyclophotocoagulation with the diode laser in contact procedure with a new focussing tip].

UNLABELLED: During contact cyclophotocoagulation (CPC) energy is transmitted through the conjunctiva and the sclera by direct contact of a plain fibre tip. Alteration and scarring of the conjunctiva and the outer parts of the sclera cannot always be avoided. We used a new fibre tip which defocuses the laser light on the conjunctiva and sclera and focuses the laser light on the ciliar body. The coupling and focusing medium between fibre and sclera is a small glass ball (3 mm diameter). METHODS: Up to now we have treated 52 human eyes of 35 patients affected by refractory glaucoma using a diode laser (Zeiss Visulas II) coupled with a 200-microns fibre and a 3-mm focusing ball tip. In all cases 20-40 spots were applied 1.5 mm posterior of the limbus, power varied between 1.5 and 2.2 W and exposure time was 1.5 s. RESULTS: In all cases the conjunctiva and the sclera was unaltered. The mean follow-up time was 224 days. The mean IOP values +/- SD before treatment and 6 days, 4 weeks and 1 year after treatment were 40.9 +/- 7.1, 19.3 +/- 7.9, 22.4 +/- 14.8 and 17.2 +/- 11.3 mmHg. No positive, linear correlation was found between number of spots and IOP drop after 6 days. There was considerable individual variance. The main complications were anterior fibrinous uveitis (10%), slight vitreous haemorrhage (2/52) and transitory hypotony (2/52). In one enucleated eye structural alterations to the pigmented and non-pigmented epithelial cells of the ciliary body were found histologically. CONCLUSIONS: Contact diode CPC can significantly reduce IOP, and the conjunctiva and sclera can be left totally unaltered by the focusing ball tip used in this study.

Aged↗

[In vitro studies on pulsed laser angioplasty in liquid and gaseous medium].

PURPOSE: In a vascular model the mechanisms of a pulsed dye laser working in a liquid and a gaseous environment was tested and documented by means of a high-speed camera. METHODS: The pulse application (630 nm wavelength) on a target of calcified arterial plaque material was detected at intervals of 20 microseconds up to 800 microseconds total time. RESULTS: In water the laser beam created a cavitation bubble which expanded from 0 to 450 microseconds from a radius 0 to 3 mm and collapsed afterwards. The average expansion velocity of the bubble was 80 m/s in the first 20 microseconds and the resulting velocity of the tube model wall was 10 m/s. Compared with gaseous atmosphere there was no bubble and consecutively no wall stress. CONCLUSION: It is obvious that laser angioplasty causes wall stress and destruction of different vascular layers. This might be the initial stimulation for restenosis. In gaseous atmosphere laser angioplasty should be more protective.

Angioplasty, Laser↗

Morphological and biomorphometrical observations on laser thermal keratoplasty. Histological and biomorphometrical examination of the relationship between refractive change and the volume following Cr:Tm:Ho:YAG laser treatment.

Laser thermal keratoplasty (LTK) is currently under clinical trial for the correction of hyperopia and hyperopic astigmatism by means of collagen coagulation in the peripheral cornea. The purpose of our study was to optimize the ratio between the volume of damaged corneal stroma and the refractive effect so as to minimize potential side effects such as endothelial damage or induction of glare phenomena. We therefore performed histological and morphometrical examinations of enucleated pig eyes to determine the relationship between the coagulated stromal volume and the refractive change after LTK using a pulsed Cr:Tm:Ho:YAG laser (wavelength 2.12 microns) on enucleated pig eyes. The refractive change was documented with a Littman ophthalmometer. Morphometrical analysis was performed using polarized light microscopy of sirius red-stained specimens. This special stain separated the thermally changed stroma into a dark nonbirefringent center and a birefringent peripheral zone. The volume of both zones was positively correlated with the refractive change induced. The volume was in turn influenced by the choice of laser parameters. From the ratio of the volume to the refractive change it was found that pulse energies above 30 mJ let to an enlargement of the coagulation volume without increasing the refractive change effectively. The use of high pulse energies did not improve the effect of LTK but only increased the risk of unwanted side effects. However, an increase in the laser repetition rate at a constant pulse number per spot led to refractive changes with minimal coagulation volume. The highest relative refractive change was achieved with a dark central zone and a birefringent zone, each having a volume of about 50-80 x 10(-3) mm3.

Animals↗

Photofragmentation of lens nuclei using the Er: YAG laser: preliminary report of an in vitro study.

The energy of the erbium: YAG laser (2.940-nm wave-length) can be used for minimally traumatic photoablation due to its high absorption at the tissue water and its consequently low penetration depth. Laser sclerostomy ab externo, an application of this principle, has undergone advanced clinical investigation. Another potential application is photofragmentation of the lens for cataract extraction. A laboratory model Er: YAG laser (flashlamp-pumped. 200-microseconds pulse length, 5-Hz repetition frequency) was coupled to a short low-OH quartz fiber (400 microns in diameter). The laser energy was applied by direct contact of the fiber tip to human lenses with very dense cataract. The lenses rested in a small cuvette filled with an aqueous-humor-analogous fluid. The fragmentablation rate was evaluated in relation to the number of pulses and the pulse energy. A laser-triggered flash-photography unit was engaged to visualize the ablation dynamics. We found tissue-ablation rates to range from 4 to 19 micrograms/pulse, depending on the nucleus density and ulse energy. The maximal size of the removed fragments was always below 500 microns. During ablation, rapidly increasing and collapsing cavitation bubbles were photographed at the distal tip of the application fiber. The impact radius of these cavitation effects markedly exceeded the pure penetration depth of laser radiation at a 2.9-microns wavelength. A clinical application of the method should be possible as judged by the results obtained for tissue-ablation rate and fragment size. Cavitation-bubble dynamics seems to be responsible for the high fragmentation efficiency. Special application probes have to be developed to optimize ablation and to prevent inadvertent destruction of the posterior lens capsule by cavitation effects.

Humans↗

In vivo uptake of liposomal benzoporphyrin derivative and photothrombosis in experimental corneal neovascularization.

BACKGROUND AND OBJECTIVE: Photodynamic therapy (PDT) has been used successfully to occlude neovascularizations experimentally. We evaluated the vasoocclusive potential of benzoporphyrin derivative (BPD), a new photosensitizer currently in clinical trials. Since liposomally formulated BPD strongly binds to endogenous low density lipoproteins (LDL) after i.v. injection, LDL act as carrier to deliver BPD preferentially to proliferating endothelial cells. STUDY DESIGN/MATERIALS AND METHODS: Corneal neovascularizations in rabbits were used as model. Time-dependent uptake and retention of liposomal BPD were measured in vivo by monitoring the laser-induced fluorescence (LIF). Photothrombosis was induced using a dye laser emitting at 692 nm. RESULTS: A maximal BPD concentration was measured at 60-90 minutes postinjection determining the optimal time interval for treatment. Exposures as low as 10 J/cm2 allowed complete and irreversible neovascular occlusion as documented angiographically. Histology revealed selective endothelial damage, adjacent corneal stroma, or iris vessels, remained intact. Identical results were obtained using BPD directly complexed with LDL suggesting use of a LDL-mediated pathway. CONCLUSION: We suggest BPD-PDT for a selective treatment of neovascular diseases.

Animals↗