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. 2007 Jan-Feb;13(1-2):22-9.
doi: 10.2119/2006-00035.Mittermayr.

Blue laser light increases perfusion of a skin flap via release of nitric oxide from hemoglobin

Rainer Mittermayr  1 , Anatoly Osipov, Christina Piskernik, Susanne Haindl, Peter Dungel, Carina Weber, Yuri A Vladimirov, Heinz Redl, Andrey V Kozlov
Affiliations

Blue laser light increases perfusion of a skin flap via release of nitric oxide from hemoglobin

Rainer Mittermayr et al. Mol Med. 2007 Jan-Feb.

Abstract

It has recently been shown that nitrosyl complexes of hemoglobin (NO-Hb) are sensitive to low-level blue laser irradiation, suggesting that laser irradiation can facilitate the release of biologically active nitric oxide (NO), which can affect tissue perfusion. The aim of this study was to evaluate the therapeutic value of blue laser irradiation for local tissue perfusion after surgical intervention. Blood was withdrawn from a rat, exposed to NO and infused back to the same rat or used for in vitro experiments. In vitro, an increase of NO-Hb levels (electron paramagnetic resonance spectroscopy) up to 15 microM in rat blood did not result in the release of detectable amounts of NO (NO selective electrode). Blue laser irradiation of NO-Hb in blood caused decomposition of NO-Hb complexes and release of free NO. Systemic infusion of NO-Hb in rats affected neither systemic circulation (mean arterial pressure) nor local tissue perfusion (Doppler blood flow imaging system). In contrast, a clear enhancement of local tissue perfusion was observed in epigastric flap when elevated NO-Hb levels in blood were combined with local He-Cd laser irradiation focused on the left epigastric artery. The enhancement of regional tissue perfusion was not accompanied by any detectable changes in systemic circulation. This study demonstrates that blue laser irradiation improves local tissue perfusion in a controlled manner stimulating NO release from NO-Hb complexes.

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Figures

Figure 1
Figure 1
Effect of RBC and laser irradiation on the levels of NO and NO-Hb in a buffered oxygen free saline solution. (A) Kinetics of NO levels upon addition of Hb and laser irradiation; (B) NO-Hb EPR spectrum; + and # indicate the features of 5- and 6- coordinated NO-Hb complexes. The magnitudes of corresponding peaks were used to estimate the amounts of both complexes; (C) NO-Hb concentration in solution before and after irradiation with a 40 mW He-Cd laser. * significantly different P < 0.01.
Figure 2
Figure 2
Kinetics of NO release and reabsorption triggered by He-Cd laser (40 mW) irradiation in erythrocytes enriched with NO-Hb. (A) Anaerobic irradiation; (B) Aerobic irradiation; (C) Difference in NO concentration in solution due to switching the laser ON and OFF.
Figure 3
Figure 3
Effect of NO-Hb on cGMP levels and MAP. (A)Effect of enriching with NO on the cGMP levels in sham blood (Sham) and in sham blood enriched NO-Hb (Sham-NO-Hb). (B) Effect of the infusion of sham and NO-Hb enriched blood on cGMP levels in the systemic circulation of rats. CON—resuscitation of sham blood; NO-Hb—resuscitation of sham blood enriched NO-Hb. (C) Effect of the infusion of sham and NO-Hb enriched blood on the MAP in systemic circulation of rats. CON—resuscitation of sham blood; NO-Hb—resuscitation of sham blood enriched NO-Hb.
Figure 4
Figure 4
Experimental protocol: surgery and monitoring of systemic circulation (MAP) and local tissue perfusion (Doppler imaging).
Figure 5
Figure 5
Effect of laser irradiation on the systemic levels of cGMP and MAP Open bars/boxes—cGMP/MAP of rats receiving sham blood, respectively. Closed bars/boxes—cGMP/MAP of rats receiving NO-Hb enriched blood, respectively.
Figure 6
Figure 6
(A) Overview of superficial perfusion of the rodent epigastric area. (B) Control group with no alterations in perfusion (C) NO-Hb Group with increase in local perfusion during laser irradiation. Circles indicate the region where the maximal changes were observed. Quantification, however, was performed for whole region displayed in the figures. 1st column—Background, 2nd column—Infusion of NO-Hb, 3rd column—Irradiation of epigastric artery, 4th column—After irradiation period.
Figure 7
Figure 7
Quantification of laser Doppler scans of the epigastric flap in the rat.
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