This study investigates the possible role of singlet oxygen in accounting for the inhibitory effect of laser-dye injury on endothelium-dependent dilations. The combination of helium-neon (HeNe) laser (20-s exposure) and intravascular Evans blue impairs endothelium-dependent dilation of mouse pial arterioles by acetylcholine (ACh), bradykinin (BK), and calcium ionophore A23187. Each has a different endothelium-derived mediator (EDRFACh, EDRFBK, EDRFionophore, respectively). In this study, diameters at a craniotomy site were monitored in vivo with an image splitter-television microscope. The laser-dye injury, as usual, abolished the responses 10 and 30 min after injury, with recovery, complete or partial, at 60 min. Dilations by sodium nitroprusside, an endothelium-independent dilator, were not affected by laser-dye. When the singlet oxygen scavengers L-histidine (10(-3) M) and L-tryptophan (10(-2) M) were added to the suffusate over the site, the responses to ACh at 10 and 30 min were relatively intact, the response to BK was partly protected at 10 min only, and the response to ionophore was still totally impaired at 10 and 30 min. Lysine, a nonscavenging amino acid, had no protective effects with any dilator. We postulate that a heat-induced injury initiates a chain of events resulting in prolonged singlet oxygen generation by the endothelial cell (not by the dye). We postulate further that destruction of EDRFACh by singlet oxygen is responsible for laser-dye inhibition of ACh and that generation of the radical must continue for > or = 30 min. On the other hand, the heat injury itself is probably responsible for the elimination of the response to ionophore. Heat plus singlet oxygen generated by heat-damaged tissue may initially impair the response to BK, but by 30 min only the effects of some other factor, presumably heat injury, account for the impaired response to BK.
"Furthermore, and still poorly appreciated in the renal microcirculation field, interactions between light and fluorophores may generate heat and photochemical processes that may affect protein synthesis in live specimens. The so-called “light-dye” or “laser-dye” effects locally and/or temporarily modify endothelial cell function (e.g., [118, 119]), and the impact of confocal microscopy on cell calcium handling and cell death has recently been emphasized in bovine chondrocyte cultures . Further studies seem therefore warranted to settle these issues linked to “classic” fluorescence, mono-, or multiphoton imaging in live tissues. "
[Show abstract][Hide abstract] ABSTRACT: Renin-producing cells have been the object of intense research efforts for the past fifty years within the field of hypertension. Two decades ago, research focused on the concept and characterization of the intrarenal renin-angiotensin system. Early morphological studies led to the concept of the juxtaglomerular apparatus, a minute organ that links tubulovascular structures and function at the single nephron level. The kidney, thus, appears as a highly "topological organ" in which anatomy and function are intimately linked. This point is reflected by a concurrent and constant development of functional and structural approaches. After summarizing our current knowledge about renin cells and their distribution along the renal vascular tree, particularly along glomerular afferent arterioles, we reviewed a variety of imaging techniques that permit a fine characterization of renin synthesis, storage, and release at the single-arteriolar, -cell, or -granule level. Powerful tools such as multiphoton microscopy and transgenesis bear the promises of future developments of the field.
[Show abstract][Hide abstract] ABSTRACT: To evaluate the possibility of oxygen radical damage in the skin after He-Ne laser irradiation according to dose intensity and time.
The He-Ne laser (lambda = 632.8, 10 mW) was used on the skin of mice with 1, 3, and 5 joule (J/cm2) dose rates for 1, 5, and 7 days in each case, and the results were compared with normal and anesthetic nonirradiated skins. The efficacy was determined by the formation of thiobarbituric acid-reactive substances (TBARS) in a 10-minute period and expressed as a concentration of malondialdehyde (MDA) from the lipid peroxidation of epidermal tissue, and total superoxide dismutases (SODs) and catalase activities, correlated with histologic biopsies.
Data from epidermal SODs, catalase activity and the degree of lipid peroxidation at low-power radiation showed that repeated exposure had led to the induction of free radical damage and of epidermal changes as confirmed by microscopic study. The application of the He-Ne laser at 1, 3, and 5 J intensity for 5 days caused a gradual increase in the SODs and catalase activities, while the levels of TBARS were slightly decreased in the mouse epidermis. However, these patterns were reversed after 3 and 5 J irradiations for 5 and 7 days laser treatment. Furthermore, microscopic examinations revealed that the laser-irradiated skin changed the release of stratum granule from epidermis to hair follicle, and produced blood vessel thrombosis of the dermal capillary plexus.
The presence of lipid peroxidation in the hairless mouse skin after exposure to He-Ne laser energy intensity of over 3 J for over 5 days was demonstrated. This lipid peroxidation could have been generated from oxygen free radicals. The histologic and oxidative enzymatic correlations on lipid peroxidation in the skin have provided a better understanding of He-Ne laser therapy-tissue interactions. It is possible to take advantage of these findings to evaluate pathologic skin conditions and effective laser dosage more efficiently.
Lasers in Surgery and Medicine 01/2000; 27(5):420-6. DOI:10.1002/1096-9101(2000)27:5<420::AID-LSM1003>3.0.CO;2-K · 2.62 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: An experimental model of cochlear ischemia using the photosensitization reaction of rose bengal (RB) is reported. Ischemia was induced by the systemic administration of RB solution and localized irradiation of the cochlea with a green light of specific wavelength for RB activation. After the RB solution was administered, the cochlear blood flow was temporarily increased and then decreased, and the latency of wave I of the auditory brainstem response increased and the amplitude of wave I decreased after RB injection and finally wave I disappeared completely. A histological study revealed thrombus formation in the vessels of the irradiated cochlea not only in the stria vascularis and the spiral ligament, but also in the modiolus. L-Histidine, a scavenger of singlet oxygen (1O2) and hydroxyl radical (.OH), significantly prolonged cochlear dysfunction, but D-mannitol, a specific scavenger of.OH did not, suggesting that 1O2 has an important role in this reaction.
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