Damage Thresholds for Exposure to NIR and Blue Lasers in an In Vitro RPE Cell System

Denton, Michael L.; Foltz, Michael S.; Estlack, Larry E.; Stolarski, David J.; Noojin, Gary D.; Thomas, Robert J.; Eikum, Debbie M.; Rockwell, Benjamin A.

doi:10.1167/iovs.05-1066

Cited by 41 publications

(27 citation statements)

References 37 publications

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“…Apart from the commonly utilized ARPE-19, there are few accounts of spontaneously immortalized RPE cell lines such as D407, RPE-J and BPEI-1 [41,42]. Additionally, it is common practice for primary cells to be transformed by viruses, such as SV40 or HPV, or other genetic manipulation such as activating a telomerase gene [28-30,43,44]…”

Section: Discussionmentioning

confidence: 99%

Isolation and characterization of a spontaneously immortalized bovine retinal pigmented epithelial cell line

2009

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BackgroundThe Retinal Pigmented Epithelium (RPE) is juxtaposed with the photoreceptor outer segments of the eye. The proximity of the photoreceptor cells is a prerequisite for their survival, as they depend on the RPE to remove the outer segments and are also influenced by RPE cell paracrine factors. RPE cell death can cause a progressive loss of photoreceptor function, which can diminish vision and, over time, blindness ensues. Degeneration of the retina has been shown to induce a variety of retinopathies, such as Stargardt's disease, Cone-Rod Dystrophy (CRD), Retinitis Pigmentosa (RP), Fundus Flavimaculatus (FFM), Best's disease and Age-related Macular Degeneration (AMD). We have cultured primary bovine RPE cells to gain a further understanding of the mechanisms of RPE cell death. One of the cultures, named tRPE, surpassed senescence and was further characterized to determine its viability as a model for retinal diseases.ResultsThe tRPE cell line has been passaged up to 150 population doublings and was shown to be morphologically similar to primary cells. They have been characterized to be of RPE origin by reverse transcriptase PCR and immunocytochemistry using the RPE-specific genes RPE65 and CRALBP and RPE-specific proteins RPE65 and Bestrophin. The tRPE cells are also immunoreactive to vimentin, cytokeratin and zonula occludens-1 antibodies. Chromosome analysis indicates a normal diploid number. The tRPE cells do not grow in suspension or in soft agar. After 3H thymidine incorporation, the cells do not appear to divide appreciably after confluency.ConclusionThe tRPE cells are immortal, but still exhibit contact inhibition, serum dependence, monolayer growth and secrete an extra-cellular matrix. They retain the in-vivo morphology, gene expression and cell polarity. Additionally, the cells endocytose exogenous melanin, A2E and purified lipofuscin granules. This cell line may be a useful in-vitro research model for retinal maculopathies.

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Section: Discussionmentioning

confidence: 99%

Isolation and characterization of a spontaneously immortalized bovine retinal pigmented epithelial cell line

2009

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“…It is well known that photons generated by a laser using a wavelength in the NIR range (IR-A range of 700–1400 nm), such as those used in translational and clinical development, may cause retinal damage if the laser probe is directed toward the eye or the beam is reflected off of a surface [98–100]. However, this type of damage depends upon a variety of factors including power density.…”

Section: Parkinson’s Diseasementioning

confidence: 99%

Transcranial near-infrared laser therapy applied to promote clinical recovery in acute and chronic neurodegenerative diseases

Lapchak¹

2012

Expert Review of Medical Devices

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One of the most promising methods to treat neurodegeneration is noninvasive transcranial near-infrared laser therapy (NILT), which appears to promote acute neuroprotection by stimulating mitochondrial function, thereby increasing cellular energy production. NILT may also promote chronic neuronal function restoration via trophic factor-mediated plasticity changes or possibly neurogenesis. Clearly, NILT is a treatment that confers neuroprotection or neurorestoration using pleiotropic mechanisms. The most advanced application of NILT is for acute ischemic stroke based upon extensive preclinical and clinical studies. In laboratory settings, NILT is also being developed to treat traumatic brain injury, Alzheimer’s disease and Parkinson’s disease. There is some intriguing data in the literature that suggests that NILT may be a method to promote clinical improvement in neurodegenerative diseases where there is a common mechanistic component, mitochondrial dysfunction and energy impairment. This article will analyze and review data supporting the continued development of NILT to treat neurodegenerative diseases.

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“…The ANSI standard [5] sets the maximum permissible exposure (MPE) as the level of laser radiation to which an unprotected person may be exposed without adverse biological changes in the eye or skin. The standard is based upon in vitro [6] and in vivo [7–11] laser bioeffects experiments designed to determine the limits of tissue to laser damage as well as to describe the photobiological processes involved [12]. Experiment exposure parameters are based on the mechanisms governing light induced tissue damage: power density, total exposure duration, and wavelength of irradiating energy.…”

Section: Introductionmentioning

confidence: 99%

High-ResolutionIn VivoImaging of Regimes of Laser Damage to the Primate Retina

Pocock

Oliver

Specht³

et al. 2014

Journal of Ophthalmology

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Purpose. To investigate fundamental mechanisms of regimes of laser induced damage to the retina and the morphological changes associated with the damage response. Methods. Varying grades of photothermal, photochemical, and photomechanical retinal laser damage were produced in eyes of eight cynomolgus monkeys. An adaptive optics confocal scanning laser ophthalmoscope and spectral domain optical coherence tomographer were combined to simultaneously collect complementary in vivo images of retinal laser damage during and following exposure. Baseline color fundus photography was performed to complement high-resolution imaging. Monkeys were perfused with 10% buffered formalin and eyes were enucleated for histological analysis. Results. Laser energies for visible retinal damage in this study were consistent with previously reported damage thresholds. Lesions were identified in OCT images that were not visible in direct ophthalmoscopic examination or fundus photos. Unique diagnostic characteristics, specific to each damage regime, were identified and associated with shape and localization of lesions to specific retinal layers. Previously undocumented retinal healing response to blue continuous wave laser exposure was recorded through a novel experimental methodology. Conclusion. This study revealed increased sensitivity of lesion detection and improved specificity to the laser of origin utilizing high-resolution imaging when compared to traditional ophthalmic imaging techniques in the retina.

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Damage Thresholds for Exposure to NIR and Blue Lasers in an In Vitro RPE Cell System

Cited by 41 publications

References 37 publications

Isolation and characterization of a spontaneously immortalized bovine retinal pigmented epithelial cell line

Isolation and characterization of a spontaneously immortalized bovine retinal pigmented epithelial cell line

Transcranial near-infrared laser therapy applied to promote clinical recovery in acute and chronic neurodegenerative diseases

High-ResolutionIn VivoImaging of Regimes of Laser Damage to the Primate Retina

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