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Biomedical subjects

M W Berns

Publications and source records attributed to M W Berns.

At least 73 records · Page 4Linked to original sources
PubMed1995

Assisted hatching in mouse embryos using a noncontact Ho:YSGG laser system.

PURPOSE: A noncontact holmium:yttrium scandium gallium garnet (Ho:YSGG) laser system has been designed and tested for the micromanipulation of mammalian embryos. The purpose of this preliminary investigation was to determine the effectiveness of this laser for assisted hatching and evaluate its impact on embryo viability. The Ho:YSGG system, utilizing 250-microsecond pulses at a wavelength of 2.1 microns and 4 Hz, was used to remove a portion of the zona pellucida (ZP) of two- to four-cell FVB mouse embryos. RESULTS: In the first experiment there was no difference in blastocyst production or hatching rates following laser or conventional assisted hatching (LAH or AH, respectively) in contrast to control embryos cultured in a 5% CO2 humidified air incubator at 37 degrees C. In the second experiment a blastocyst antihatching culture model was employed and LAH-treated embryos were cultured in a serum-free HTF medium (HTF-o). Blastocyst formation was not influenced by LAH treatment and hatching was increased (P < 0.01) from 4 to 60% compared to HTF-o control group. CONCLUSIONS: These preliminary data demonstrate the utility and nontoxic properties of the Ho:YSGG laser system for quick and precise ZP drilling.

Animals↗
PubMed1995

Evidence for localized cell heating induced by infrared optical tweezers.

The confinement of liposomes and Chinese hamster ovary (CHO) cells by infrared (IR) optical tweezers is shown to result in sample heating and temperature increases by several degrees centigrade, as measured by a noninvasive, spatially resolved fluorescence detection technique. For micron-sized spherical liposome vesicles having bilayer membranes composed of the phospholipid 1,2-diacyl-pentadecanoyl-glycero-phosphocholine (15-OPC), a temperature rise of approximately 1.45 +/- 0.15 degrees C/100 mW is observed when the vesicles are held stationary with a 1.064 microns optical tweezers having a power density of approximately 10(7) W/cm2 and a focused spot size of approximately 0.8 micron. The increase in sample temperature is found to scale linearly with applied optical power in the 40 to 250 mW range. Under the same trapping conditions, CHO cells exhibit an average temperature rise of nearly 1.15 +/- 0.25 degrees C/100 mW. The extent of cell heating induced by infrared tweezers confinement can be described by a heat conduction model that accounts for the absorption of infrared (IR) laser radiation in the aqueous cell core and membrane regions, respectively. The observed results are relevant to the assessment of the noninvasive nature of infrared trapping beams in micromanipulation applications and cell physiological studies.

Animals↗
PubMed1995

Root canal preparation using the second harmonic KTP:YAG laser: a thermographic and scanning electron microscopic study.

Thermal and microstructural events resulting from KTP laser use during root canal preparation were investigated in 30 extracted single-rooted human teeth. In the first section of this study, thermal events occurring on the root surfaces of 18 teeth during and after exposure of the root canal were measured using thermography. A variety of parameters were used to determine settings that would be effective without causing thermal damage to the periodontal ligament. In the second section of the study, root canals of 12 teeth exposed to KTP laser irradiation at parameters derived from section 1 were evaluated using Scanning electron microscopy. KTP laser application at a power setting of 3 W, an exposure time of 2 s, and a frequency of 5 Hz, applied five times, removed smear layer and debris from the root canal surface at temperatures below the thermal injury threshold for periodontal tissue.

Argon↗
PubMed1995

Photomedicine of the endometrium: experimental concepts.

Gynaecological photomedicine offers new diagnostic and therapeutic methods based on the interaction of light with the reproductive organs. One example is photodynamic therapy (PDT) in which photosensitizers are applied systemically or topically for selective endometrial ablation. Several studies describing the potential use of PDT for this application are reviewed. Basic experimental and clinical aspects of PDT, such as photosensitizer types, application modes, irradiation parameters, optical properties of tissues and photodegradation of photosensitizers are discussed.

Animals↗
PubMed1995

Rat reproductive performance following photodynamic therapy with topically administered Photofrin.

A rat animal model was used for comparing the photodynamic efficacy of two formulations of topically administered Photofrin in the uterus: 0.7 mg/kg Photofrin and 0.7 mg/kg Photofrin + 4% Azone, a penetration-enhancing agent. Uterine structure and reproductive performance were evaluated following illumination with 80 J/cm2 of 630 nm light. Fluorescence microscopy was employed to determine drug localization in frozen uterine sections at various times after drug administration. Functionality studies demonstrated a significant reduction in the number of implantations per treated uterine horn compared to controls. The mean number of implantations decreased systematically on increasing the interval between Photofrin administration and light application. At 72 h, 0.88 +/- 0.52 gestational sacs per rat were recorded with Photofrin therapy, compared with 8.1 +/- 1.12 (P = 0.01) on the untreated side, indicating nearly complete loss of reproductive capability. Similar results were achieved after only 3 h treatment with Photofrin + Azone (0.38 +/- 0.26 sacs per rat versus 7.5 +/- 1.07 on the untreated side; P = 0.01). This indicates that the effect of Photofrin can be enhanced either by extending the drug incubation period from 3 to 72 h or by adding the penetration-enhancing drug Azone. Fluorescence pharmacokinetic studies suggest that both forms of topically administered Photofrin are diffusely distributed throughout the endometrium at virtually the same rate. However, Azone may enhance the selectivity of photodynamic therapy by facilitating drug targeting to critical endometrial structures.

Administration, Topical↗
PubMed1995

Intrauterine light delivery for photodynamic therapy of the human endometrium.

Photodynamic therapy is currently being evaluated as a minimally invasive procedure for endometrial ablation not requiring anaesthesia. Light penetration depths at 630, 660 and 690 nm and the optimal configuration of intrauterine light-diffusing fibres were determined in 14 human uteri to assist in the design of a light intrauterine device. Post-menopausal ex-vivo uteri showed a significantly lower light penetration depth than pre-menopausal uteri. With a single central diffusing fibre inserted, the fluence rate measured in the uterine wall at the most remote point of the cavity decreased to 1.1 +/- 0.4% of that measured at closest proximity, whereas it decreased to only 40.0 +/- 9.0% with three fibres. Distension of the uterine cavity with 2 ml of an optically clear fluid increased the fluence rate at the fundus between the fibres at a depth of 2 mm by a factor of 4. We conclude that in normal-sized pre-menopausal uterine cavities, three diffusing fibres will deliver an optical dose above the photodynamic threshold level at a depth of 4 mm, even in the most remote areas, in < 30 min without causing thermal damage. For distorted and elongated cavities, either slight distension of the cavity or the insertion of a fourth diffusing fibre is required.

Adult↗
PubMed1995

Developmental competence of mouse embryos following zona drilling using a non-contact holmium:yttrium scandian gallium garnet (Ho:YSGG) laser system.

The purpose of this study was to assess the efficacy of the holmium:yttrium scandian gallium garnet (Ho:YSGG) laser, operating in a pipette-free, non-contact mode, to assist hatching and sustain normal embryonic development. Two-cell mouse embryos were recovered and assigned to laser-assisted hatching (LAH) treatment or control human tubal fluid (HTF) culture with or without serum (HTF-s, HTF-o) or with late serum supplementation (HTF-o/s). The basic experimental apparatus for LAH consisted of a stationary 2.1 microns Ho:YSGG laser beam directed through a mechanical shutter into an input port of a Zeiss Axiomat inverted microscope. Fewer (P < 0.05) embryos developed to the blastocyst stage in the HTF-s group (81%) than in the LAH (90%), HTF-o (94%) and HTF-o/s (92%) groups. The level of hatching was significantly increased (P < 0.01) after the LAH treatment (57%) compared to HTF-o/s (32%), HTF-s (18%) or HTF-o (5%). Implantation rates were not significantly impaired following the LAH treatment (21%). These data demonstrate that LAH using the Ho:YSGG laser is a simple, accurate and effective procedure for assisted hatching.

Animals↗
PubMed1995

Autofluorescence spectroscopy of optically trapped cells.

Cellular autofluorescence spectra were monitored in a single-beam gradient force optical trap ("optical tweezers") in order to probe the physiological effects of near infrared and UVA (320-400 nm) microirradiation. Prior to trapping, Chinese hamster ovary cells exhibited weak UVA-excited autofluorescence with maxima at 455 nm characteristic of beta-nicotinamide adenine dinucleotide (phosphate) emission. No strong effect of a 1064 nm NIR microbeam on fluorescence intensity and spectral characteristics was found during trapping, even for power densities up to 70 MW/cm2 and radiant exposures of 100 GJ/cm2. In contrast to the 1064 nm trap, a 760 nm trapping beam caused a two-fold autofluorescence increase within 5 min (about 20 GJ/cm2). Exposure to 365 nm UVA (1 W/cm2) during 1064 nm trapping significantly altered cellular autofluorescence, causing, within 10 min, a five-fold increase and a 6 nm red shift versus initial levels. We conclude that 1064 nm microbeams can be applied for an extended period without producing autofluorescence changes characteristic of alterations in the cellular redox state. However, 760 nm effects may occur via a two-photon absorption mechanism, which, in a manner similar to UVA exposure, alters the redox balance and places the cell in a state of oxidative stress.

Animals↗
PubMed1995

Effect of ND:YAG laser irradiation and root planing on the root surface: structural and thermal effects.

Effects of ND:YAG laser irradiation on untreated and root planed tooth roots were investigated to determine whether a cleaning effect and/or removal of smear layer could be achieved without concomitant microstructural or thermal damage. Sixty (60) healthy extracted teeth were either untreated, irradiated only, root planed only, or irradiated and root planed. Intra-pulpal and surface temperatures were monitored during irradiation, then SEM was performed. Smear layer elimination was achieved without inducing hard tissue microstructural damage at 5W, using pulse durations and intervals of 0.1 s, a fluence of 0.77 J/cm2, and a total energy density of approximately 700 J/cm2. However, these results were not consistent in all samples. At these parameters, intra-pulpal temperature increases of 9 to 22 degrees C and surface temperature increases of 18 to 36 degrees C were recorded. Thus, despite their effectiveness for smear layer removal, these parameters may not be appropriate for clinical use as an adjunct to conventional periodontal therapy.

Body Temperature↗
PubMed1995

Photodynamic therapy of ocular melanoma with bis silicon 2,3-naphthalocyanine in a rabbit model.

PURPOSE: To investigate bis (tri-n-hexylsiloxy) silicon 2,3-naphthalocyanine (SINc; 0.5 mg/kg) for photodynamic therapy of an experimental ocular melanoma in pigmented rabbits. METHODS: SINc was dissolved in canola oil by heating, emulsified with Tween 80, and administered by ear vein. Pharmacokinetics were studied in frozen tumor sections by fluorescence microscopy using a charge coupled device, camera-based, low-light detection system with digital image processing at 1 and 24 hours. A Ti:sapphire laser and a microlens were used to deliver the light (770 nm; 40 mW/cm2; 20 J/cm2). A control rabbit received light without SINc. RESULTS: Localization studies of SINc showed intravascular distribution shifting to a tumor stromal and perivascular distribution 24 hours after treatment. Tissue thermal damage after irradiation was minimal in the control. Exudative retinal detachments were not observed. Tumor destruction was observed, with sharp demarcation to a depth of 3.5 mm. CONCLUSIONS: Tumor light penetration was good at 770 nm, and thermal effects from the exciting light alone were minimal. Photodynamic therapy with SINc resulted in localized tumor destruction reflecting the light beam path without damage to adjacent tissue or intraocular complications.

Animals↗
PubMed1995

Effect of freezing on the relative escape force of sperm as measured by a laser optical trap.

OBJECTIVE: To determine the possible effect of freezing on sperm escape force as measured by laser trap. DESIGN: Controlled clinical study. SETTING: Normal volunteers, academic setting. PATIENTS: Normal, healthy volunteers. INTERVENTION: Sperm selection, freezing and thawing. Measurement of relative (sperm) escape force before and after freezing. MAIN OUTCOME MEASURE: Comparison of escape force as a measurement of freezing effects. RESULTS: Wide individual variations noted, with some individuals showing significant increases or decreases. The relative escape force in fresh (76.1 +/- 31.1 mW) and in frozen-thawed samples (75.6 +/- 40.0 mW) were similar. CONCLUSION: Freezing does not affect the relative escape force of normal sperm. Any possible detrimental effect of freezing on sperm may be related to other sperm functions.

Freezing↗
PubMed1994

Directed movement of chromosome arms and fragments in mitotic newt lung cells using optical scissors and optical tweezers.

A pulsed-laser microbeam at 532 nm wavelength (optical scissors) and a laser-induced optical trap at 1064 nm wavelength (optical tweezers) have been successively combined to dissect and manipulate chromosomes in live newt lung epithelial cells. These preliminary experimental results demonstrated that chromosome fragments dissected by laser microbeam surgery, regardless of their size, could be easily pulled or rotated by optical forces when positioned at the periphery of the mitotic spindle. In addition, chromosome arms which were not subjected to laser microsurgery also could be moved with the optical tweezers at the spindle periphery. In our previous study on rat kangaroo kidney cells (PTK2), this degree of facilty in manipulating chromosome movement was not possible, most likely due to the close proximity of the intermediate filament "cage" to the spindle. It is concluded herein that optical scissors and tweezers can be used in combination to study the interaction of chromosomes with the mitotic spindle in cells where the peripheral regions of the spindle are unobstructed by intermediate filaments. This can be performed on newt cells, where the diameter of the cage can be substantially larger than the diameter of the spindle.

Animals↗
PubMed1994

Photodynamic destruction of endometrial tissue with topical 5-aminolevulinic acid in rats and rabbits.

OBJECTIVE: The goal of this study was to determine the optimal parameters for photodynamic endometrial destruction with topically applied 5-aminolevulinic acid, a precursor for the endogenous synthesis of the fluorescent photosensitizer protoporphyrin IX. STUDY DESIGN: 5-Aminolevulinic acid pharmacokinetics were measured in rat and rabbit models by analyzing tissue frozen sections 3 to 12 hours after topical administration. Dose-response studies were conducted for 100 to 400 mg/ml 5-aminolevulinic acid. Photodynamic therapy was performed intraluminally, and tissue morphologic features were evaluated 3 and 7 days after treatment. RESULTS: Peak fluorescence was observed 3 hours after topical administration. Glandular fluorescence significantly exceeded stromal and myometrial in all studies, particularly for 200 mg/ml 5-aminolevulinic acid. Histologic studies revealed persistent epithelial destruction with minimal regeneration. CONCLUSION: Topical 5-aminolevulinic acid photodynamic therapy can be used for highly effective, long-lasting destruction of endometrial epithelium. However, optical dosimetry can vary, particularly in the rabbit model, and this appears to have an impact on long-term reepithelialization.

Administration, Topical↗
PubMed1994

Differential vascular response to laser photothermolysis.

Individual blood vessels in the chick choriallantoic membrane were selectively coagulated through photothermolysis, using pulsed laser irradiation at 585 nm. Pulse durations were chosen to be 0.45 ms and 10 ms, which correspond to the thermal relaxation times in blood vessels of 30 microns and 150 microns diameter, respectively. The short pulses, at a light fluence F = 3 Jcm-2, caused permanent occlusion of vessels of 40 microns diameter or less, whereas larger caliber vessels (60-120 microns) required F = 4-5 Jcm-2. The long-duration pulses, at F = 7 Jcm-2, caused coagulation of the larger diameter vessels; the small-caliber vessels and capillaries showed resistance to photothermolysis and required multiple exposures to achieve coagulation. The fluence versus diameter (F versus d) relationship for coagulation was calculated for the two pulse durations. The energy deposited in a cylindrical absorber of diameter d by an optical field, incident perpendicular to the vessel, was expressed analytically and compared with the energy required to coagulate a blood vessel of the same lumen dimeter. When thermal diffusion is incorporated into the model, our findings can be accounted for quantitatively. This information will be of use for improving the laser treatment of port wine stains and other vasculopathies. A surprising observation was that arterioles were damaged at lower incident energy densities than venules having the same lumen diameter, despite the fact that absorbance in oxygen-rich and oxygen-poor blood is the same at 585 nm.

Allantois↗
PubMed1994

Benzoporphyrin derivative: a potent photosensitizer for photodynamic destruction of rabbit endometrium.

OBJECTIVES: To determine the optimal pharmacokinetic characteristics for photodynamic endometrial destruction using topically applied benzoporphyrin derivative and to evaluate long-term morphologic effects in a rabbit model. METHODS: Benzoporphyrin derivative pharmacokinetics were measured by analyzing frozen tissue sections 1.5-12 hours following topical administration. Photodynamic therapy was induced intraluminally 1.5 hours after drug application, and tissue morphology was evaluated by light and scanning electron microscopy 1 and 4 weeks after treatment. RESULTS: The highest glandular and stromal fluorescence was observed 1.5 hours after topical administration. Relative fluorescence of the endometrial glands significantly exceeded that of stroma and myometrium mainly at 1.5 and 3 hours (P < .0001). Histology revealed persistent epithelial destruction with minimal regeneration. Stroma resembled scar tissue in the majority of sections. The bordering myometrium was loosened and invaded by connective tissue. CONCLUSION: Topically applied benzoporphyrin derivative can be used for highly effective, long-lasting photodynamic destruction of rabbit endometrium. However, optical dosimetry can vary, particularly in the rabbit model, and this appears to affect long-term reepithelialization.

Administration, Topical↗
PubMed1994

Relative force of human epididymal sperm.

OBJECTIVE: To assess the relative escape force of human epididymal sperm using a laser generated optical trap and compare it with that of human ejaculated sperm. DESIGN: Evaluation of the relative force generated by epididymal and ejaculated sperm using an 800-nm laser-generated optical trap system (titanium-sapphire, model 899-01; Coherent Innova, Palo Alto, CA). SETTING: University-based facility at the Beckman Laser Institute and Medical Clinic and Center for Reproductive Health, University of California, Irvine. INTERVENTIONS: A total of 2,720 sperm from 28 samples were randomly analyzed. Fifteen were ejaculated samples (1,650 sperm) obtained from men with proven fertilization, and 13 were epididymal samples (1,070 sperm) aspirated microsurgically from patients with obstructive azoospermia. An optical trap equipped with the 100x Neofluar objective was used to analyze an average of 100 sperm per patient. MAIN OUTCOME MEASURES: Determination of mean relative escape force values in milliwatts for epididymal and ejaculated sperm samples. RESULTS: The mean relative escape force for epididymal sperm was 32.4 mW, significantly lower than ejaculated sperm, which was 85.1 mW. By correlating epididymal sperm relative force with fertilization in vitro at an arbitrary cutoff value of 30 mW, it was found that no fertilization occurred if a sample had < 13% of sperm at that value. CONCLUSIONS: [1] The average relative escape force of the epididymal sperm was found to be 60% weaker than that of ejaculated sperm. [2] It is demonstrated that the noncontact laser optical trap is a sensitive tool that can evaluate single sperm force as a new physiological parameter.

Ejaculation↗