Current laser safety standards for multiple pulse lasers are based primarily on modeling and the results of single pulse
studies. Previous thermal effects studies have focused on histological and visible endpoints, with only a few studies
examining the actual temperatures achieved. The goal of this research was to probe the actual vertical temperature profile
produced by 2.01 micron laser pulses in the cornea. In this study the corneal temperature rise from multiple 2.01 micron
Tm:YAG laser pulses was investigated using ex-vivo rabbit eyes. A thermal-measurement data set for a different number
of pulses was collected and compared. An infrared thermal camera employing microbolometer detectors captured surface
temperature rises resulting from laser pulses. Single 10 ms pulses as well as two, three, and four pulse sequences were
utilized while the total energy delivered was held constant. A comparison of the data to temperatures required for
denaturing proteins and the current laser safety guidelines will be presented.
Human perception of 2.0-µm infrared laser irradiation has become significant in such disparate fields as law enforcement, neuroscience, and pain research. Several recent studies have found damage thresholds for single-pulse and continuous wave irradiations at this wavelength. However, the only publication using multiple-pulse irradiations was investigating the cornea rather than skin. Literature has claimed that the 2.0-µm light characteristic thermal diffusion time was as long as 300-ms. Irradiating the skin with 2.0-µm lasers to produce sensation should follow published recommendations to use pulses on the order of 10 to 100 ms, which approach the theoretical thermal diffusion time. Therefore, investigation of the heating of skin for a variety of laser pulse combinations was undertaken. Temperatures of ex vivo pig skin were measured at the surface and at three depths from pulse sequences of six different duty factors. Differences were found in temperature rise per unit exposure that did not follow a linear relation to duty factor. The differences can be explained by significant heat conduction during the pulses. Therefore, the common heat modeling assumption of thermal confinement during a pulse may need to be experimentally verified if the pulse approaches the theoretical thermal confinement time.
Non-contact temperature measurement and imaging instruments are widely used in studies of laser interaction
with tissue. For reliable results, independent verification of the instruments abilities and limitations is necessary. Two
common types of heat measuring instruments are the LiTaO3 type II pyrometer and the microbolometer array thermal
imager. This study found that when considering the Signal to Noise Ratio, the temporal resolution, and the spatial
dependency of the temperature measured by the instrument the microbolometer was superior for measurements of
incident beams of 5 mm diameter and all pulse frequencies investigated.
The determination of sensation thresholds has applications ranging from uses in the medical community such as
neural pathway mapping and for the diagnosis of diabetic neuropathy, to potential uses in determining safety standards.
This study sought to determine the sensation threshold, and the distribution of sensation probabilities, for pulse trains
ranging from two 10 ms pulses to nine 10 ms pulses from 2.01 μm laser light incident on a human forearm and chest.
Threshold was defined as the energy density that would elicit sensation 50% of the time (ED50). A method of levels
approach was used in conjunction with a monovariate binary response model to determine the ED50. We determined the
ED50 and also a distribution of threshold probabilities. Threshold was found to be largely dependant on total energy
deposited for smaller pulse trains, and thus independent of the number of pulses. Total energy becomes less important as
the number of pulses increases however, and a decrease in threshold was measured for a nine pulse train as compared to
one through four pulse trains. Thus we have demonstrated that this method is a useful and easy way for determining
sensation thresholds from a 2.01 μm laser for possible clinical use. We have also demonstrated that lower power lasers
when pulsed can elicit sensation at comparable levels to higher power single pulse lasers.
Laser alignment thermal sensitive paper can be used for laser applications such as spot size measurements, beam
characteristics, and determining beam mode. Thermal sensitive paper interactions with a 1.54 micron, 35 ns, 3.75 J, Er-Glass laser produced spots that had three concentric zones of response. These spots interact with each other if a
minimum distance between exposures is not maintained. The distance needed between spots is directly related to the
energy density incident upon the paper. Although there was no lot designation for the box of thermal sensitive papers
used in this research, we were able to determine that the response of different papers in the same box could vary under
constant laser exposure parameters. Therefore, investigators need to be cautious when extrapolating experimental data
from exposures using this type of laser alignment paper.
KEYWORDS: In vitro testing, Tissues, Data modeling, Injuries, Laser tissue interaction, Cornea, In vivo imaging, Eye models, Animal model studies, Laser damage threshold
Despite the increasing use of infrared lasers in medical, industrial, and military settings, data on threshold radiant exposures and median effective dose (ED50) as they relate to laser-tissue interaction are limited. Our goals were to determine the ED50 for single-pulse, 1540-nm laser exposures in ex vivo and in vitro rabbit corneal models and to characterize the histopathological changes associated with the laser-tissue interaction. An erbium-glass laser was used to deliver single, 1540-nm wavelength pulses to 27 ex vivo and 24 in vitro rabbit corneal models. The ex vivo model was exposed to single pulses of 0.8-ms duration and radiant energies ranging from 17.61 J/cm2 to 42.26 J/cm2. The in vitro corneal models were exposed to single pulses of 0.8 ms duration and had radiant exposures ranging from 14.87 to 29.72 J/cm2. Tissue exposure sites were observed for presence of a lesion immediately post-exposure and 24 h after exposure. Histopathological evaluations of tissue exposure sites were conducted 24 h after exposure. The ED50 was determined to be 21.24 J/cm2 for the in vitro rabbit corneal models and 30.86 J/cm2 for the ex vivo corneal models. Both the in vitro and ex vivo models displayed similar histopathological responses of tissue necrosis and epithelial cell proliferation.
Background and purpose: The purpose of this study is to determine the impact of melanin on skin response to single 3.8 micron, eight microsecond laser pulses and the difference in lesion formation thresholds for input into laser safety standards. Williams et al., performed a study examining laser tissue interaction from 3.8-micron lasers in lightly pigmented Yorkshire pigs (Sus scrofa domestica). However, studies performed by Eggleston et al comparing pigmented and lightly pigmented skin with human skin found that the Yucatan mini-pig is a superior model for laser skin exposures.
Methods: Five Yucatan mini-pigs under general anesthesia were exposed to 3.8 micron laser pulses ranging from 0.8 J/cm2 to 93 J/cm2. Gross examinations were done acutely and 24 hours after laser exposure. Skin biopsies were then collected at various times post exposure, and histologic examinations were conducted. Results: The 24 hour ED50 was determined to be 4.5 J/cm2 with fiducial limits of 6.2 and 2.2 J/cm2. As deposited energy was increased, the lesion presentation ranged from whitening of the epidermis (4 J/cm2) to whitening with inflammatory centers (14 J/cm2), and at the highest energy levels inflammatory areas were replaced with an epidermal ulcerated central area (>21 J/cm2).
Conclusion: Preliminary findings suggest pigmentation or melanin may play a minor role in the mechanism of laser-tissue damage. The ED50 of Yorkshire pigs was 2.6 J/cm2. The ED50 of the Yucatan mini-pig was found to be 3.6 J/cm2, and although it was higher, it is still within the 95% fiducial limits.
As a consequence of the enormous expansion of laser use in medicine, industry and research, specific safety standards must be developed that appropriately address eye protection. The purpose of this study is to establish injury thresholds to the cornea for 3.8 micron 8 microsecond laser light pulses and to investigate a possible replacement model to live animal testing. Previous studies of pulsed energy absorption at 3.8 microns were performed using rhesus monkey cornea and were at pulse durations two orders of magnitude different than the 8 microsecond pulses used in this study. Ex-vivo pig eyes were exposed at varying energies and evaluated to establish the statistical threshold for corneal damage. Histology was used to determine the extent of damage to the cornea. It is expected that the results will be used to assist in the establishment of safety standards for laser use and offer an alternative to future animal use in establishment of safety standards.
The goal of this study is to determine if a high energy laser pulse can cause internal injury that cannot be grossly visualized. High power lasers are currently in development such as the Medical Free Electron Laser (MFEL), the Anti-Ballistic Laser (ABL) and the Tactical High Energy Laser (THEL) and the potential exists for human exposure. Little is known about the effects of these high output lasers on internal organs when a thoracic exposure occurs. This study utilized a 3.8 micron single 8 microsecond pulse laser for all exposures. Yucatan miniature pigs were exposed to a single pulse over the sternum. In addition, some animals were also exposed in the axillary region. Creatine phosphokinase (CPK) and troponin levels were measured prior to and post exposure to assess cardiac muscle damage. Gross and histologic changes were determined for the porcine skin, lung tissue, and cardiac muscle. This study explores if a greater than class 4 laser classification is warranted based on the potential for thoracic injury.
The purpose of this study was to evaluate the laser-tissue interactions of engineered human skin and in-vivo pig skin following exposure to a single 3.8 micron laser light pulse. The goal of the study was to determine if these tissues shared common histologic features following laser exposure that might prove useful in developing in-vitro and in-vivo experimental models to predict the bioeffects of human laser exposure. The minimum exposure required to produce gross morphologic changes following a four microsecond, pulsed skin exposure for both models was determined. Histology was used to compare the cellular responses of the experimental models following laser exposure. Eighteen engineered skin equivalents (in-vitro model), were exposed to 3.8 micron laser light and the tissue responses compared to equivalent exposures made on five Yorkshire pigs (in-vivo model). Representative biopsies of pig skin were taken for histologic evaluation from various body locations immediately, one hour, and 24 hours following exposure. The pattern of epithelial changes seen following in-vitro laser exposure of the engineered human skin and in-vivo exposure of pig skin indicated a common histologic response for this particular combination of laser parameters.
Laser induced breakdown has the lowest energy threshold in the femtosecond domain, and is responsible for production of threshold ocular lesions. It has been proposed that multiphoton absorption may also contribute to ultrashort-pulse tissue damage, based on the observation that 33 fs, 810 nm pulse laser exposures caused more DNA breakage in cultured, primary RPE cells, compared to CW laser exposures delivering the same average power. Subsequent studies, demonstrating two-photon excitation of fluorescence in isolated RPE melanosomes, appeared to support the role of multiphoton absorption, but mainly at suprathreshold irradiance. Additional experiments have not found a consistent difference in the DNA strand breakage produced by ultrashort and CW threshold exposures. DNA damage appears to be dependent on the amount of melanin pigmentation in the cells, rather than the pulsewidth of the laser; current studies have found that, at threshold, CW and ultrashort pulse laser exposures produce almost identical amounts of DNA breakage. A theoretical analysis suggest that the number of photons delivered to the RPE melanosome during a single 33-fsec pulse at the ED50 irradiance is insufficient to produce multiphoton excitation. This result appears to exclude the melanosome as a locus for two- or three-photon excitation; however, a structure with a larger effective absorption cross-section than the melanosome may interact with the laser pulses. One possibility is that the nuclear chromatin acts as a unit absorber of photons resulting in DNA damage, but this does not explain the near equivalence of ultrashort and CW exposures in the comet assay model. This equivalence indicated that multiphoton absorption is not a major contributor to the ultrashort pulse laser damage threshold in the near infrared.
Two-dimensional electrophoresis and histomorphometry were used to determine if equivalent protein changes occurred within native rabbit corneas and engineered corneal tissue models following in vitro exposure to single pulse, 1540 nm laser light operating at a pulse width of 0.8 milliseconds. Frozen sections of exposed tissues were processed to detect laser-induced protein changes. Isoelectric points, molecular weights and relative densities were used to characterize corneal proteins of interest that were then identified using MALDI-MS peptide fragment analysis. Increasing radiant exposures of corneal tissues were associated with progressively more severe necrosis of the epithelium and stroma in both the native and engineered tissues.
The purpose of this review is to compile information on the optical and healing properties of the cornea when exposed to infrared lasers. Our long-term goal is to optimize the treatment parameters for corneal injuries after exposure to infrared laser systems. The majority of the information currently available in the literature focuses on corneal healing after therapeutic vision correction surgery with LASIK or PRK. Only a limited amount of information is available on corneal healing after injury with an infrared laser system. In this review we will speculate on infrared photon energy absorption in corneal injury and healing to include the role of the tear layer. The aim of this review is to gain a better understanding of infrared energy absorption in the cornea and how it might impact healing.
There are several data sources for collecting laser incidents. All reviewed sources collect information differently for varying purposes. An effort was undertaken to combine laser exposure reporting data into a single database so that trends in laser incidents could be identified. A review of available datasets revealed significant disparities in laser exposure reporting. As a result, utilizing the existing database to predict personnel at increased risk for laser exposure and injury is challenging if not impossible. For example, many of the data sources do not contain information about physical examinations, diagnosis, or medical follow-up, which are important for studying laser injury outcomes. This study proposes using the Delphi Technique to identify relevant fields that should be collected for a laser incident database based on the experiences of three groups of United States Air Force (USAF) professionals: (1) Engineers (Bioenvironmental Engineers), (2) Health Physicists, and (3) Physicians (Ophthalmologists and Flight Surgeons). In broad terms, these three professional groups coordinate laser incident analyses and investigations. Knowing what information is most important for studying laser incidents is the first step in establishing an effective database that will assist in identifying occupations that are at high-risk for laser injury. Robust data sets obtained for analysis by these healthcare professionals can be an effective tool for laser injury prevention and management.
Five male Yorkshire pigs were exposed on their flank to 4 microsecond pulses of laser light from a Deuterium Fluoride 3.8 micron Laser at varying energies. A preliminary ED50 threshold for various skin reactions was determined for this laser exposure combination. The animal’s skin was assessed for injury immediately, 1 hour, 24 hours and 72 hours post exposure. In general, energies below 3.2 J/cm2 leave no lasting skin reaction. As energy increased above the threshold, erythema or skin reddening was easily visualized. High-energy pulses appear to produce a “rug burn” erythema without evidence of punctate hemorrage (bleeding) or coagulation. Laser exposure sites on the pigs were also biopsied to obtain histopathological results. These findings suggest that the principal effect of this type of in-vivo laser exposure is removal of the epithelium, while not damaging the papillary dermis or structures beneath the Basement Membrane Zone (BMZ).
Yucatan mini-pigs and Yorkshire pigs were exposed on their flanks to 1318 nm, 0.5 ms laser pulses. Injuries were readily visible on the Yorkshire pigs immediately, one hour, and 24 hours post exposure but difficult to locate at 3 days post exposure. The Yucatan mini-pig injuries were not seen immediately or at one-hour post exposure, but at 24 hours and three days post exposure they were easily identified. The Yorkshire injuries were round red, well demarcated, with a circular pink area of edema. It is hypothesized that skin pigmentation has an effect on the mechanism of 1318 nm laser energy absorption in skin. Pigmentation may have a significant effect on how infrared laser injuries present, develop and heal.
An increasing number of industries, to include military, medicinal, and technological arenas, are using 1.3 micron laser systems for which current skin and eye guidelines are identical. No skin threshold, ED50 or exposure data are available. The mechanisms of laser-tissue interaction with skin at 1.3 microns are unknown. Together, these facts necessitate increased research to prevent future laser accidents and injuries. This study examines the method of interaction of 1.3 microns laser light with tissue in the Yorkshire pig. Our research addresses laser-tissue interaction through delivery using a Nd:YAG with an intracavity filter producing 1.3 micron light at 0.5 millisecond exposure time and in the range of 137 to 475 J/cm2. Laser exposure to Yorkshire pigs was evaluated for dermal lesion development. Lesions were appraised for acute, one-hour and 24-hour post exposure presentation.
Yucatan Mini-pigs were exposed on their flank to 0.5 milli second 1318 nm pulses of laser light. The ED50 damage threshold was determined for this laser exposure combination. The skin was assessed for injury immediately, at 1 hour, 24 hours and three days post exposure. Generally, at least 24 hours was required for visible lesions to form. It was found that as the duration between exposure and assessment expanded the injury was more easily visualized. Tissue samples were collected for histology at one hour, 24 hours and three days. Histologic sections will be presented in future work. It was also found that the topical application of mineral oil to the area of interest was found to greatly increase the ease of identification of injuries.
Recently a review of multiple laser injury and accident databases was initiated. Last year we reported on preliminary findings in this area. This past year the search for laser injury and accident reports was expanded, and a significant number of additional laser incidents were located. The database from the Food and Drug Administration’s Center for Devices and Radiological Health was supplemented with more up to date information and non-medical laser incidents were added. Rockwell Laser Industries database was verified to not contain duplicates from the new information, as was the Army’s Laser Accidents and Incidents Registry. Information from the Federal Aviation Administration regarding laser accidents and incidents were also included. Incidents not resulting in laser specific injuries has been tracked. This information was not included in our previous report. In this study, case reports are used to show gross trends in laser injury, accident and incident reporting. This study is still in progress, and evaluation is incomplete.
In this study reports of laser injuries in all three military services (Air Force, Army and Navy/Marine) are compared. In collecting data for this study laser injury reports provided by Rockwell Laser Industries (RLI), the United States Army Medical Research Detachment of the Walter Reed Army Institute of Research which maintains the Laser Accident and Incident Registry (LAIR), the Food and Drug Administration's Center for Devices and Radiological Health (CDRH), and the three service Safety Centers. We found a total of 29 laser injury reports that met our case definition. Since 1965, when the first injury occurred, there have been 6 Air Force, 15 Army, and 8 Navy/Marine injuries reported. Statistical analysis of data analyzed thus far shows no difference between the services in 8-year risk groupings between 1965-2002.
In vitro exposures of explant rabbit corneas to single pulse 1540 nm infrared laser light operating at a pulse width of 0.8 milliseconds resulted in coagulative necrosis of both the corneal epithelium and stroma. Histomorphometric data correlated with increasing tissue radiant exposures. Histologic alterations in the corneal stroma were typical of matrix remodeling within the beam path and reactive to antibodies against matrix metalloproteinase-2. A two-dimensional electrophoretic analysis, using a mini-gel format, was developed to determine if specific corneal protein changes within tissue sections could be detected. Frozen sections taken through the center of the laser lesion were evaluated for proteomic data using tissue isoelectric focusing in the first dimension and polyacrylamide gel electrophoresis in the second dimension. Histomorphometric data describing the extent of the laser lesions were compared to the isoelectric points, molecular weights and relative densities of individual corneal proteins. Increasing radiant exposures of corneal tissues were associated with characteristic histomorphometric and proteomic changes.
These milestone papers tell the story of effects previously thought to be impossible to produce in tissue. The studies evolve from speculation to experimentation to theory, and culminate in policy recommendations. This comprehensive reference book documents the initial probing limits of subnanosecond pulses and the resulting discoveries of nonlinear effects.
Significant damage to rabbit corneal tissue was produced by a single pulse, in vitro exposure of 1540 m infrared laser light operating in either millisecond or nanosecond pulse widths. Millisecond pulse widths of infrared laser light produced a marked coagulative necrosis of both the corneal epithelium and stroma. We also noted histologic alterations in the stromal matrix within the beam path that we interpreted as matrix remodeling. To test this interpretation, we used an indirect immunohistochemical procedure to detect Matrix Metalloproteinase-2 (MMP-2) activity. Immunohistochemistry revealed that the MMP-2 reaction was mostly limited to the margins of the beam path. In addition, the MMP-2 reaction was less intense than expected given the significant tissue changes observed in the histologic sections. Exposure of rabbit corneal tissue to the nanosecond pulse widths produced a less severe coagulative necrosis of the tissue when compared to the millisecond exposures. However, a markedly stronger immunohistochemical pattern than would have been predicted from the histologic sections was observed, with approximately half of the beam path filled with MMP-2 reaction product. These data suggest an association between infrared laser pulse width and the degree of extracellular matrix remodeling in rabbit corneal tissue.
Threshold, median effective dose, and the mechanism of laser-tissue interaction are not well defined at the 1318-nm wavelength for human corneal exposures. The goals of this research effort are to identify at-risk groups, characterize the lesions imposed, and establish the ED50 for single pulse 1318-nm laser exposures on the cornea. A Neodymium: Yttrium Aluminum Garnet (Nd:YAG) laser was used to deliver 1318-nm wavelength pulses to the corneas of ten female Dutch Belted rabbits (Oryctolagus cuniculus). Single pulses of 0.5-ms duration and radiant beam energy ranging from 116 to 2250 joules/per square centimeter (J/cm2) were used. Exposure sites were clinically evaluated acutely, one hour and twenty-four hours post-exposure for the presence of a lesion. Results from the twenty-four hour evaluation were used to determine the ED50. Grossly, the lesions appeared as small, circular, well-demarcated, white, opaque lesions. Histologically, the lesions appeared as conical shaped coagulative necrosis with the base of the lesion at the epithelial surface of the cornea and extending to the apex at the endothelial border of the cornea. The ED50 for 1318-nm exposures to the rabbit cornea was determined to be 383 J/cm2 for a 0.1-mm spot size as measured at 1/e2.
Laser use is pervasive and steadily expanding both in the private sector and the Department of Defense (DoD). For more than 20 years, Rockwell Laser Industries, the U.S. Army, and the Food and Drug Administration's Center for Devices and Radiological Health have separately collected data on injuries occurring during, or resultant from, the use of lasers. However, data from these sources is incomplete and has not recently undergone a thorough compiling, statistical analysis, review and summarization. It is our belief that in order to evaluate current related medical surveillance, safety and training procedures, this data needs such an examination. Persons maintaining these databases were contacted and any available data on laser injury was collected. The data was analyzed and examined for pertinent similarities and differences among a wide range of parameters. We summarize these findings in this paper and also comment on the injuries, current safety measures and injury reporting protocols associated with laser use.
Our work addresses laser tissue interaction for skin exposures from 1318 nm laser pulses. Single pulse data from 0.5 milli-second exposures, along with the mechanisms of photon energy absorption in tissue are investigated. We offer preliminary ED50 data and its implication within the realm of laser tissue interaction for discussion. A comparison will be made between the skin reaction of the Yucatan mini-pig (highly pigmented model) and Yorkshire pig (lightly pigmented model). This study represents the first systematic histological investigation of skin reaction to 1318 nm laser pulses.
Completion of recent studies within our group indicates a breed-based difference in dermal response to 1540 nm 0.8 millisecond laser pulses. Laser exposure to Yucatan Mini- Pigs (highly pigmented skin) and Yorkshire pigs (lightly pigmented skin) demonstrate statistical differences between the ED50's of the two breeds. Laser delivery is accomplished using an Er:Glass system producing 1540 nm of light at millisecond exposure times and in the range of 5 to 95 J/cm2. Dermal lesion development was evaluated for acute, 1 hour, and 24-hour post exposure presentation. Our data contradicts the theory that water absorption is the sole mechanism of dermal tissue damage observed from 1540 nm laser exposures, as skin chromophores appear to play a role in lesion development.
High-energy use of 1318 nm laser systems is becoming more prevalent in military and industrial settings. Threshold, ED50, exposure data and mechanism of laser-tissue interaction need to be determined for this wavelength using appropriate animal models that allow for extrapolation to control human exposures. Threshold, ED50, exposure data at 1318 nm for retinal and corneal injury have previously been undertaken and reported for rhesus monkeys. Using comparable methods, we examine exposure data at 1318 nm to determine the ED50 and laser-tissue interaction in the rabbit model to evaluate cornea. We present preliminary data for the ED50 threshold on the cornea from exposure to 1318 nm single laser pulses. Delivery of laser energy is accomplished using an Nd:YAG system producing 1318 nm light in the 0.5 millisecond time exposure regime and in the range of 0 to 500 mJ. Results from this work will aid in the establishment of safety standards for near infrared laser systems.
Single pulse, 1540 nm laser light with a pulse width of 1 microsecond altered the morphologic appearance of explant rabbit and pig corneas following ex vivo exposure. Using digital images of the post-exposure corneas projected onto a measuring grid, we could accurately locate the relative position of the circular laser lesion in the embedded tissue. This allowed us to section through the lesion with micrometer precision and accurately resolve the inside edge, middle and outside edge of the laser lesion. All tissue sections used for morphometric analysis were taken through the middle of the lesion. Several features of the response to laser exposure may reflect species-specific tissue differences. The rabbit corneal epithelium showed a homogeneous coagulative necrosis with a distinct demarcation between necrotic and normal epithelium. The pig epithelium also showed a distinct demarcation between necrotic and normal epithelium, however, there were several remarkable differences in the tissue response between the two species including coagulative necrosis pattern and nuclear morphology. These changes suggested a different and less severe response of the pig epithelium to the laser light when compared to the rabbit epithelium.
Dutch Belted rabbit corneas and corneal equivalent (CE) tissue were exposed to 0.8 millisecond pulses of 1540 nm laser light. We report the single pulse ED50 for Dutch belted rabbits and for in-vitro corneal equivalent tissues. A histological comparison between the two tissues is presented. Remarkable similarities between the two models in both location and extent of damage are noted. We postulate which cellular energy absorption mechanisms are significant at 1540 nm and how this relates to the histopathology presented.
Mechanisms of tissue damage are investigated for skin and cornea exposures from 1540 nm ('eye safe') laser single pulses of 0.8 milli-seconds. New skin model data point out the advantages of using the Yucatan mini-pig versus the Yorkshire pig for in-vivo skin laser exposures. Major advantages found include similarities in thickness and melanin content when compared with human skin. Histology from Yucatan mini-pig skin exposures and the calculation of an initial ED50 threshold indicate that the main photon tissue interaction may not be solely due to water absorption. In-vitro corneal equivalents compared well with in-vivo rabbit cornea exposure under similar laser conditions. In-vivo and in-vitro histology show that initial energy deposition leading to damage occurs intrastromally, while epithelial cells show no direct injury due to laser light absorption.
The military and industry are using 1540 nm laser systems for which current consensus safety standards are misleading. Threshold, ED50, exposure data, along with mechanisms of laser-tissue interaction need to be more accurately determined. Recent studies within our group indicate the Yucatan mini-pig is a more applicable animal model for laser induced skin injury investigators. Laser delivery is accomplished using an Er:Glass system producing 1540 nm of light at millisecond exposure times and in the range of 17 to 77 J/cm2. Dermal lesion development is evaluated for acute, 1 hour, and 24 hour post exposure presentation. Preliminary data obtained from dermal exposures indicate a difference in ED50 for Yorkshire and Yucatan pigs. In the Yucatan mini-pig erythematous lesions are formed acutely while in the Yorkshire, lesions are seen at 24 hours. Preliminary data indicates that lesion development occurs at or near the basal layer of the epidermis causing nuclear pyknosis, cellular swelling and loss of cellular detail. Contrary to the theory that water absorption is the primary mechanism of dermal tissue damage observed from 1540 nm laser exposures, skin chromophores appears to play a role in lesion development.
We investigate the use of in-vitro corneal equivalents as a replacement model for in-vivo rabbit corneas used in laser damage threshold studies. In-vitro corneal equivalents (CE) were exposed to 0.8 millisecond (ms), 1540 nanometer (nm) single laser light pulses ranging from 124 J/cm2 to 58 J/cm2. After exposure, CE's were evaluated opthalmoscopcopically, imaged using confocal microscopy and examined histologically to investigate the mechanisms of laser induced damage. Preliminary results indicate that the 50% damage threshold for CE's, ED50, is approximately 70 J/cm2 with a 0.5 mm diameter spot size. Additional data points are required to determine a statistically significant ED50. Preliminary exposures of in-vivo rabbit corneas using the same laser parameters as the CE's are also reported. Comparisons between the in-vivo and in-vitro models are presented. Histopathological images from both models show remarkable similarities in the location and extent of damage throughout the full thickness of each tissue.
Current safety standards for lasers operating in the 1400 to 10,000 nm wavelength region are based on few observations at specific wavelengths using in vivo models that may not represent an accurate correlation to human integument. Based on experimental results conducted with Yorkshire pigs, these standards may not accurately reflect the potential for laser injury when humans are exposed to these wavelengths. It is our belief that one of the primary damage mechanisms involved in these laser injuries is due to energy absorption by skin pigmentation, or melanin. Qualitatively, Yorkshire pigs lack melanin in their skin when compared to a more highly pigmented animal, such as the Yucatan minipig. It is hypothesized that the Yucatan minipig is a more appropriate model for pigmented human skin. By comparing histologic samples taken from various locations on Yucatan minipigs and Yorkshire pigs, and comparing these to potential locations of skin exposure on humans, we present a discussion for the establishment of more appropriate locations for in vivo laser exposure studies.
We report on preliminary work undertaken to determine ED50 thresholds for both skin and cornea exposure from 1400 to 2000 nm laser light. Work presented here is focused initially on 1540 nm exposures to both human skin and cornea. Light microscopy and confocal microscopy used to help understand the type of photon-tissue interactions responsible for skin and corneal injury are discussed along with preliminary results from these techniques. Further, we report on in vivo models which are considered to best represent human skin for laser tissue interaction studies. Additionally, in vitro models for corneal exposure are discussed as replacement models for in vivo corneal exposures.
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