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Biomedical subjects

M W Berns

Publications and source records attributed to M W Berns.

At least 55 records · Page 3Linked to original sources

Photodynamic therapy (PDT) of the ciliary body with silicon naphthalocyanine (SINc) in rabbits.

BACKGROUND AND OBJECTIVE: To investigate silicone naphthalocyanine (SINc; 0.5 mg/kg) for photodynamic therapy (PDT) of the ciliary body in pigmented rabbits. STUDY DESIGN/MATERIALS AND METHODS: SINc was dissolved in canola oil by heating, emulsified with Tween-80, and given by ear vein. Pharmacokinetics were studied in frozen sections by fluorescence microscopy using a CCD camera-based, low light detection system with digital image processing at 1 hr and 24 hr (12 rabbits, 24 eyes total). A Ti:Sapphire laser delivered light at 770 nm by contact fiberoptic (1,000 microns; 80 mW/cm2;20,40 and 80 J/cm2). Controls (5 rabbits), received laser light at 770 nm without SINc. For comparison, eyes received continuous wave Nd:YAG laser by fiberoptic contact (0.8-1.2 J). RESULTS: Localization studies showed intravascular distribution shifting to a ciliary body distribution at 24 hr. PDT at 1 hr and 24 hr postinjection showed a more selective destruction of the ciliary body at 24 hr. Ciliary processes treated at 24 hr showed infarction and marked edema with sparing of iris. Tissue thermal damage was minimal in PDT controls. Eyes treated with the Nd:YAG laser exhibited full-thickness thermal necrosis of iris, ciliary processes, and a fibrinous iridocyclitis. In contrast, eyes treated by PDT were quiet with thrombosis of superficial blood vessels. CONCLUSION: Tissue photon penetration is good at 770 nm and thermal effects from the exciting laser alone were minimal. The ciliary processes of pigmented rabbits exhibit a selective retention of SINc and on that basis can be selectively destroyed with a minimum on thermal damage to nontarget tissues.

Aluminum Oxide↗

Dosimetry model for photodynamic therapy with topically administered photosensitizers.

BACKGROUND AND OBJECTIVE: Photodynamic therapy (PDT) based on topical application of photosensitizers has been under development over the last years. Typical applications are treatment of basal cell carcinoma of the skin and photoablation of the endometrium. The dosimetry for topically administered photosensitizers must take a time-dependent inhomogenous drug distribution into account together with the conventional parameters such as optical scattering, absorption, and photobleaching. STUDY DESIGN/MATERIALS AND METHODS: This study presents a dosimetry model where the cytotoxic dose is calculated in a stepwise procedure. The first step calculates the time-dependent distribution of 5-aminolevulinic acid (5-ALA) from diffusion theory. In skin this distribution is dependent on drug permeability through the stratum corneum, on the diffusivity of dermis and epidermis, on the drug clearance time, and on the conversion rate from 5-ALA to protoporphyrin IX (PpIX). In the second step the distribution of PpIX is calculated from the 5-ALA distribution found in the first step taking the dynamics of the biosynthesis of 5-ALA to PpIX and the clearance time of PpIX into account. In the third step the generation of cytotoxic singlet oxygen is calculated from the optical distribution during irradiation, taking a photobleaching mechanism into account. RESULTS: The distribution of cytotoxic oxygen is predicted from the optical dose, the drug dose, and the time between the application of the drug and the irradiation. CONCLUSION: The presented dosimetry model is made as simple as possible, yet composite enough to enable all relevant parameters to be taken into account. The model that is based on a linear theory in a semi-infinite medium can, if required, be extended to take nonuniform and nonlinear phenomena into account.

Administration, Topical↗

Animal model for thoracoscopic laser ablation of emphysematous pulmonary bullae.

BACKGROUND AND OBJECTIVE: Thoracoscopic laser techniques have been described for treatment of pulmonary bullae. Clinical application of this procedure has proliferated despite limited data regarding efficacy or optimal techniques. The objective of this study was to develop an animal model for investigating laser treatment of bullous lung disease. STUDY DESIGN/MATERIALS AND METHODS: Sixty-two New Zealand White rabbits (3-5 kg) were injected intravenously with 0.35 cc sterile-filtered Sephadex G-100 beads (1 g/100 cc suspension). Three hours later, rabbits were anesthetized, intubated, and 10 cc 0.7% heat-treated or 1% untreated carrageenan solution was instilled endotracheally into a catheter wedged in a mainstem bronchus. RESULTS: Bullae formed over 4-6 weeks in 33% of the animals treated with 0.7% heat-treated carrageenan, and 90% of animals receiving 1% untreated carrageenan (P < 0.005) as demonstrated by serial thoracoscopy. Thoracoscopy was performed at 6-8 weeks using 5 mm trocars under general anesthesia and mechanical ventilatory support. Animals developed pulmonary bullae ranging in size from 0.5 to 2 cm. Bullae were ablated under thoracoscopic visualization using a CO2 laser with a 4 mm OD rigid probe and short focal length in a defocused mode, or an Nd:YAG laser with a 0.4 mm diameter flexible fiberoptic probe. Animals recovered quickly following thoracoscopy. CONCLUSION: We have successfully developed an animal model for thoracoscopic laser ablation of emphysematous pulmonary bullae. This animal model should be useful in investigating treatment of bullous lung disease in humans.

Animals↗

Photosensitization of the rat endometrium following 5-aminolevulinic acid induced photodynamic therapy.

BACKGROUND AND OBJECTIVE: The impact of photodynamic therapy (PDT) on the endometrium following topical application of 5-aminolevulinic acid (ALA) was studied in a rat model. Study Design/ MATERIALS AND METHODS: Fluorescence microscopy revealed peak ALA to protoporphyrin IX (Pp IX) conversion 3-6 hours after application. Significantly higher Pp IX levels were observed in the endometrial glands compared with endometrial stroma and myometrium. RESULTS: Histological studies showed PDT-induced endometrial destruction with atrophy 7-10 weeks after treatment. Reproductive performance studies demonstrated significant implantation failure in the treated uterine horns compared with controls. The number of implantation sacs in the treated and untreated horns was 0.4 +/- 0.3 and 8.9 +/- 1.0, respectively (P < 0.01). CONCLUSION: We conclude that the high rate of Pp IX conversion in the endometrial glands can be exploited to treat dysfunctional uterine bleeding with PDT. In addition, this concept may also be applied to study endometrial regeneration and embryo implantation mechanisms.

Administration, Topical↗

Photodynamic effects on human and chicken erythrocytes studied with microirradiation and confocal laser scanning microscopy.

BACKGROUND AND OBJECTIVE: Photodynamic therapy (PDT) of cancers is associated with the destruction of the microvasculature supplying the tumor. The study elucidates the role of red blood cells in PDT-induced vascular injury. STUDY DESIGN/MATERIALS AND METHODS: Intracellular accumulation of several photosensitizers in human (non-nucleated) and chicken (nucleated) erythrocytes, as well as photodynamic induced hemolysis were studied using 488 nm laser microirradiation (15 microW) and confocal laser scanning fluorescence microscopy. RESULTS: Cells incubated with anionic hydrophilic compounds TPPS4 and Pd-TPPS4 exhibited no fluorescence before irradiation, but developed strong and sustained fluorescence in the cellular and nuclear membranes following photoinduced membrane damage. In contrast, microirradiation of Photofrin-incubated erythrocytes showed instantaneous fluorescence which decreased due to photodegradation. For the cationic hydrophilic dye methylene blue, significant fluorescence was detected only in the nucleus. Following ALA incubation, large intercellular differences were observed in fluorescence in the red spectral region. Photofrin induced the most efficient hemolysis. Higher radiant exposures were required for lysis of nucleated rather than of non-nucleated red blood cells, except in the case of methylene blue. CONCLUSION: Laser microbeams were used, for the first time, to study photodynamic cell damage. Erythrocytes were shown to be primary targets in PDT. Damage to red blood cells could be responsible for hemostasis in the vascular bed of a tumor, which was reported by many groups.

Animals↗

Structural and functional effects of endometrial photodynamic therapy in a rat model.

OBJECTIVE: Our purpose was to determine the optical dose required for irreversible endometrial destruction and prevention of implantation by photodynamic therapy with topical 5-aminolevulinic acid. STUDY DESIGN: Three hours after drug application 74 female Sprague-Dawley rats received varying doses of 630 nm of light delivered by an intrauterine cylindric diffusing fiber. RESULTS: A 64 J/cm2 in situ optical dose resulted in long-term irreversible endometrial destruction; 43 J/cm2 damaged endometrial stroma and myometrium but not glandular epithelium 1 day after photodynamic therapy. At this lower light dose endometrium regenerated to full thickness within 3 weeks; however, implantation sacs were significantly reduced. CONCLUSIONS: Photodynamic destruction of glandular epithelium accompanies irreversible endometrial ablation, whereas isolated stromal damage leads to reproductive impairment only. The optical dose required for endometrial ablation is approximately 1.5-fold higher than for reproductive impairment (functional damage) because of differential cell photosensitivity.

Aminolevulinic Acid↗

Selective photosensitizer distribution in vulvar condyloma acuminatum after topical application of 5-aminolevulinic acid.

OBJECTIVE: Our purpose was to determine the feasibility of selective photosensitization of vulvar condylomas by use of tropical application of 5-aminolevulinic acid. STUDY DESIGN: In vivo fluorescence was assessed and biopsy specimens of condylomas were taken for fluorescence microscopy in 24 patients at different times after application of 2.5% 5-aminolevulinic acid ointment or 20% 5-aminolevulinic acid cream. RESULTS: Both in vivo fluorescence imaging and fluorescence microscopy showed selective fluorescence of condylomas of the labia minora and vestibule only within short time intervals, because fluorescence of poorly keratinized normal epithelium was induced by both 5-aminolevulinic acid formulations. In non-hair-bearing skin, lesional fluorescence remained highly selective. Fluorescence microscopy showed that 90 minutes after drug application peak selectivity in epithelial lesional fluorescence was significantly higher with 2.5% 5-aminolevulinic acid ointment (4.5 +/- 0.9) than it was with 20% cream (2.1 +/- 0.2). CONCLUSION: Selective fluorescence of vulvar condyloma acuminatum can be induced by nonselective topical 5-aminolevulinic acid application. Studies evaluating selective photodynamic destruction of condylomas are justified.

Administration, Topical↗

Physiological monitoring of optically trapped cells: assessing the effects of confinement by 1064-nm laser tweezers using microfluorometry.

We report the results of microfluorometric measurements of physiological changes in optically trapped immotile Chinese hamster ovary cells (CHOs) and motile human sperm cells under continuous-wave (CW) and pulsed-mode trapping conditions at 1064 nm. The fluorescence spectra derived from the exogenous fluorescent probes laurdan, acridine orange, propidium iodide, and Snarf are used to assess the effects of optical confinement with respect to temperature, DNA structure, cell viability, and intracellular pH, respectively. In the latter three cases, fluorescence is excited via a two-photon process, using a CW laser trap as the fluorescence excitation source. An average temperature increase of < 0.1 +/- 0.30 degrees C/100 mW is measured for cells when held stationary with CW optical tweezers at powers of up to 400 mW. The same trapping conditions do not appear to alter DNA structure or cellular pH. In contrast, a pulsed 1064-nm laser trap (100-ns pulses at 40 microJ/pulse and average power of 40 mW) produced significant fluorescence spectral alterations in acridine orange, perhaps because of thermally induced DNA structural changes or laser-induced multiphoton processes. The techniques and results presented herein demonstrate the ability to perform in situ monitoring of cellular physiology during CW and pulsed laser trapping, and should prove useful in studying mechanisms by which optical tweezers and microbeams perturb metabolic function and cellular viability.

2-Naphthylamine↗

Wavelength dependence of cell cloning efficiency after optical trapping.

A study on clonal growth in Chinese hamster ovary (CHO) cells was conducted after exposure to optical trapping wavelengths using Nd:YAG (1064 nm) and tunable titanium-sapphire (700-990 nm) laser microbeam optical traps. The nuclei of cells were exposed to optical trapping forces at various wavelengths, power densities, and durations of exposure. Clonal growth generally decreased as the power density and the duration of laser exposure increased. A wavelength dependence of clonal growth was observed, with maximum clonability at 950-990 nm and least clonability at 740-760 nm and 900 nm. Moreover, the most commonly used trapping wavelength, 1064 nm from the Nd:YAG laser, strongly reduced clonability, depending upon the power density and exposure time. The present study demonstrates that a variety of optical parameters must be considered when applying optical traps to the study of biological problems, especially when survival and viability are important factors. The ability of the optical trap to alter either the structure or biochemistry of the process being probed with the trapping beam must be seriously considered when interpreting experimental results.

Animals↗

Doppler grid surface scanning applications for pulmonary subsurface parenchymal perfusion assessment.

Subsurface perfusion to lung parenchyma underlying the pleura is difficult to assess in live ventilated animals. The purpose of this study was to assess applicability of a newly developed laser Doppler grid scanning imaging technology that measures perfusion of pleural subsurface lung regions in intact normal and abnormal animal lungs. Eighty-six Doppler grid perfusion measurements were performed in five New Zealand White Rabbits (3-5 kg); four with unilateral bullous lung disease, one normal control. Left upper lobe lung surface was exposed to 10 1-sec spot Nd:YAG exposures (70 W/cm2). One week following laser exposure, all rabbits underwent sequential bilateral open thoracotomy. Unaffected left lower lobes in these animals and all four lobes of a previously untreated rabbit were used as controls. Pleural subsurface perfusion measurements were recorded over a contiguous 900-pixel square surface grid using quantitative noncontact laser Doppler imaging during open thoracotomy procedures. Scans were obtained in a normal volume ventilation mode, at 30 cm of inspiratory hold airway pressure, and postinflation. A perfusion-pressure response curve was obtained in normal lung at 10-, 20-, and 30-cm static airway pressure. Post mortem measurements were used as 0 flow controls. Normal lung tissue was found to have relatively high pleural subsurface perfusion (1362 +/- 328 corrected units on a scale of 0-4095). Areas of atelectasis had decreased perfusion (659 +/- 512 U., 48.4 +/- 12.5% compared to normal lung, p < 0.02), but returned to normal levels after inflation of the lung (1253 +/- 363 U., p = 0.21 compared to normal). Pleural subsurface perfusion decreased uniformly and progressively as lung inflation pressure increased (p < 0.0001). Perfusion increased immediately to supranormal values following release of high inspiratory inflation pressure holds (1603 +/- 626 U., 117 +/- 18% compared to normal lung, p = 0.03). Bullae had markedly decreased perfusion (541 +/- 68 U.) that was not further reduced by increased inflation pressures. Noncontact laser Doppler grid perfusion imaging appears to provide a new tool for measuring pleural subsurface perfusion over a large area of lung surface in clinical experimental settings. Results are rapid, reproducible, and consistent. Sampling errors inherent in current point sampling Doppler flow techniques are reduced by the multiple contiguous measurements. We have used this technique to demonstrate inspiratory pressure-related reduction in pleural subsurface perfusion in normal lung, reversible decreased perfusion in atelectatic regions, and reduced perfusion in bullous and laser-treated lung regions.

Analysis of Variance↗

Identification of photoacoustic transients during pulsed laser ablation of the human temporal bone: an experimental model.

Laser ablation of hard tissues during neurotologic operations has been accomplished with continuous-wave (CW) lasers in the visible and midinfrared spectrum. The mechanism of ablation at these wavelengths is secondary to photothermal-induced tissue destruction. As a result, significant thermal damage to surrounding tissue may occur. Pulsed ultraviolet (UV) lasers have been suggested as an alternative to the argon, KTP-532, and CO2 lasers currently used in clinical practice. The pulse length of Excimer lasers are considerably shorter than the thermal diffusion time of bone tissue, and as a consequence thermal injury is minimal. This makes pulsed lasers an attractive tool for tissue ablation in the ear: in essence a "cold knife." However, the short pulse width of Excimer lasers (typically 10-150 ns) can create large thermoelastic stresses in the ablation specimen. This study identifies the presence of these photoacoustic waves during the Excimer laser treatment of the cadaveric human temporal bone. A XeCl (lambda = 308 nm, tau p = 12 ns) excimer laser was used to ablate hard tissue surrounding the oval window and facial ridge with energies of 75, 45, 25, and 12 mJ/pulse. Spot size was estimated to be 0.5 mm2. Custom high-frequency polyvinyldifluoride (PVDF) piezoelectric film transducers were fabricated and attached to the promontory, round window niche, and facial ridges. The signals were amplified using a low-noise preamplifier and recorded on a digitizing oscilloscope. Photoacoustic waves were clearly identified. Notably, large acoustic waves were measured on the promontory and on both sides of the facial ridge. The implications and clinical relevance of these findings is discussed and compared to findings obtained from a model system.

Acoustics↗

Effects of ultraviolet exposure and near infrared laser tweezers on human spermatozoa.

Photostress has to be considered during optical micromanipulation of gametes. Ultraviolet light, including low-energy UVA (32-400 nm) radiation, as well as high-intensity near infrared (NIR) laser radiation may induce cell damage. A total number of 580 light-exposed sperm cells were studied in single-cell photostress experiments. Low-power (1.5 mW, 5.3 W/cm2) UVA exposure with 365 nm radiation of a standard mercury microscopy lamp to human spermatozoa resulted within 109 +/- 30 s in paralysis and within 310 +/- 110 s in cell death. Cytotoxic effects during cell manipulation with laser microbeams were found to be partly based on non-linear excitation phenomena, in particular two-photon absorption by endogenous cell chromophores. Two-photon absorption will be more intense in the case of pulsed laser microradiation, but occur also during micromanipulation with highly focused continuous wave (cw) microbeams used as laser tweezers ('optical traps'). In particular, short-wavelength NIR traps < 800 nm induce UVA-like biological effects (oxidative stress). For example, sperm trapping with 760 nm microbeams resulted in UVA-like autofluorescence modifications, paralysis within 35 +/- 20 s and cell death within 65 +/- 20 s. In contrast, laser microbeams at 800-1064 nm may act as relatively safe micromanipulation tools. In most optical traps multifrequency cw lasers are employed. Radiation of these lasers can magnify cytotoxic effects. Therefore, single-frequency laser operation should be preferred. In general, laser assisted cell micromanipulation requires a new understanding of microbeam-cell interaction, including aspects of non-linear optics.

Cell Survival↗

Regeneration processes in rabbit endometrium: a photodynamic therapy model.

The origin and process of regeneration in rabbit endometrium was evaluated following photodynamic epithelial destruction using topically applied aminolevulinic acid (ALA). Selective destruction of endometrial epithelium was performed using photodynamic therapy (PDT). ALA was diluted to 200 mg/ml dextran 70 shortly prior to administration. A volume of 1.2 ml was injected into the left uterus. Intrauterine illumination (wavelength 630 nm, light dose 40-80 J/cm2) was performed 3 h after drug administration. Tissue morphology was evaluated by light and scanning electron microscopy 1, 3, 7 and 28 days post-treatment (three animals at each time-point). Regeneration of the endometrium following epithelial ablation by PDT was fully activated after 24 h and was completed after 72 h. Endometrial surface generation occurred by proliferation, originating primarily in deeper regions of the glands. Findings from our morphological follow-up study support the origin of endometrial regeneration being mainly from undifferentiated stem cells and residual glandular epithelium.

Aminolevulinic Acid↗

CO2 and Nd:YAG laser-induced pulmonary parenchymal lung injury in a rabbit model.

Laser exposure of the pulmonary parenchyma during treatment of emphysema and other clinical indications causes acute lung injury. Animal investigations are needed to understand and control laser-induced lung injury. We hypothesized that lung injury is deeper from Nd:YAG laser exposures than CO2 exposures because of deeper penetration of Nd:YAG wavelength light. We compared the temporal evolution of histologic injury in rabbits resulting from continuous mode shallow CO2 and Nd:YAG laser pulmonary parenchymal exposures applied in rabbits. Forty-six New Zealand white (NZW) rabbits underwent treatment with CO2 laser (n=18), Nd:YAG laser (n=18), or sham thoracotomy control (n=10) to the visceral pleural surface using 1 min of exposure (5 watts, defocused to 70 W/cm2 power density for both lasers). Animals were killed at 0, 4, 7, 21, and 49 d after exposure. Lung injury, similar to that seen clinically in humans, developed in all laser-treated animals. Injury progressed from ischemia and vascular congestion, to edema and necrosis, followed by pleural and parenchymal fibrosis. The acute injury was qualitatively distinct and slightly deeper in CO2 than Nd:YAG-treated animals (p<0.02) despite the shallower depth of penetration of the CO2 laser. These findings may imply that higher absorption coefficient for CO2 laser energy results in greater focal temperatures and injury in the areas of direct exposure, and suggest that Nd:YAG laser exposure at these settings may cause shallower injury than CO2 lasers in humans undergoing clinical treatment.

Absorption↗

Determination of motility forces of human spermatozoa using an 800 nm optical trap.

The measurement and calculation of trapping forces on ellipsoidal specimens, and the determination of intrinsic motility forces of human spermatozoa by employing an 800 nm optical trap ("laser tweezers"), are presented. ATP-driven motility forces were calculated from calibrated trapping forces generated during the interaction of an 800 nm laser beam with single sperm cells. Calibration was performed by moving optically trapped sperm heads through a laminar fluid and calculating viscous forces for an ellipsoidal cell shape. Sperm heads were obtained by microsurgically removing flagellum with a pulsed laser beam ("laser scissors"). A trapping efficiency of 0.12 +/- 0.02 and a mean intrinsic motility force of 44 +/- 20 pN were determined for motile spermatozoa from healthy donors.

Humans↗

Thermal effects of CO2 laser on the pulpal chamber and enamel of human primary teeth: an in vitro investigation.

BACKGROUND AND OBJECTIVE: Thermal effects of direct exposure of the pulp chamber to CO2 laser irradiation were evaluated and potentially safe parameters for its clinical application were determined. STUDY DESIGN/MATERIALS AND METHODS: Using conventional methods, the pulp chambers of freshly extracted primary cuspids were exposed and then subjected to CO2 laser irradiation at the following parameters: 2, 4, 6, 8, 10, 15, and 20 Watts power settings; 0.76 mm diameter spot size; total energy densities between 4-1,058 J/cm2; exposure time between .01-60 s. Exposures in the continuous wave mode were performed in some specimens. In others, single firings of a .01 s pulse duration and a 1 s pulse interval were conducted. Temperature measurements were conducted using a thermocouple and thermal camera. RESULTS: Significant statistical differences in maximum temperature rise on enamel were observed in groups that received total energy densities > or = 8 J/cm2 (P < or = 0.01); within the pulp chambers these were observed in groups that used > or = 44 J/cm2 (P < or = 0.01). CONCLUSION: Parameters appropriate for pulpotomy were observed at energy densities up to 176 J/cm2 in the continuous wave mode and 264 J/cm2 using single pulses, wherein temperature rises below 5.5 degrees C were recorded on tooth surface.

Body Temperature↗

Photodynamic laser cyclodestruction with chloroaluminum sulfonated phthalocyanine (CASPc) or Photofrin (PII) vs. Nd:YAG laser cyclodestruction in a pigmented rabbit model.

BACKGROUND AND OBJECTIVE: To investigate Photofrin (PII) and CASPc for photodynamic therapy (PDT) of the ciliary body in rabbits. STUDY DESIGN/MATERIALS AND METHODS: PII (10 mg/kg) or CASPc (1 mg/kg) was given by ear vein. Pharmacokinetics were studied in frozen sections by fluorescence microscopy (CCD camera based low light detection system with digital image processing) at 1 and 24 h (8 rabbits; 16 eyes). Laser light was delivered (argon pumped dye laser; 630 and 675 nm; 8 rabbits; 16 eyes) by contact fiberoptic. To compensate for iris attenuation, irradiance was 125 mW/cm2 (20, 40, 80, or 160 J/cm2). Controls (4 rabbits; 8 eyes) received laser light without photochemicals (OD) and for comparison, continuous wave Nd:YAG laser by fiberoptic (0.8-1.2J; OS). RESULTS: Localization studies showed intravascular distribution with some selective ciliary body distribution at 24 h (PII > CASPc). Rabbits treated with PII or CASPc exhibited variable amounts of gross ciliary body edema, infarction, and necrosis by 24-48 h. This response was not seen in PDT control tissues; damage was seen in the iris and ciliary body, with partial vacuolization of the pigment epithelium. CONCLUSION: PDT may offer a more selective approach to ciliary body destruction. A small but significant thermal effect was seen during PDT from melanin photon uptake with damage to iris and ciliary body. Thermal damage and potential interaction with ocular visual pigments may limit use of these photochemicals and wavelengths for PDT of the ciliary body.

Animals↗