Semi Quantitative Assessment of Inducible NO Synthase am


Published on

Theralase Technologies Inc. designs, develops, manufactures and markets patented, superpulsed laser technology utilized in biostimulation and biodestruction applications. The technology is safe and effective in the treatment of chronic pain, neural muscular-skeletal conditions and wound care. When combined with its patented, light-sensitive Photo Dynamic Compounds, Theralase laser technology is able to specifically target and destroy cancers, bacteria, viruses as well as microbial pathogens associated with food contamination.

Published in: Health & Medicine, Technology
  • Be the first to comment

  • Be the first to like this

No Downloads
Total views
On SlideShare
From Embeds
Number of Embeds
Embeds 0
No embeds

No notes for slide

Semi Quantitative Assessment of Inducible NO Synthase am

  1. 1. 13/12/2011 Semi -Quantitative Assessment of Inducible NO Synthase in an Acute Inflammation Model Yumi Moriyama1, Jamie Fong 2 , Arkady Mandel 2 , Margarete K. Akens 3 , Lothar Lilge 1,4 1 Ontario Cancer Institute, Princess Margaret Hospital, Toronto, Canada, 2 Theralase Inc. Toronto, Canada, 3 Sunnybrook and Women’s College Health Centre., Toronto, Canada 4 Medical Biophysics, University of Toronto, Toronto, CanadaAbstract: Methodology TABLE. Comparison of Sum of Integral of BLI as a Function of ResultsWhile various mechanisms of action for Low Level Ages in Different LLLT ExposuresLaser Therapy have been proposed, it is evident that 30 transgenic iNOS-luc ( FVB/N-Tg) mice Controlthe photon energy delivered to the tissue is (n=10) and Treated (n=30) startified as young < Histological analysis (l=635nm) Groups < 15 week 15-32 weeks > 1year P-valueinsufficient as an energy source for any biological 15 weeks versus old > 1 year.outcome and needs to be augmented by biological Control 9.97 – 3.7 5.65 – 4.7 3.42-1.43 0.021*processes to improve ATP production or similar. M 635 1.57 – 3.9 3.28 – 4.3 15.2-8.9 0.02*While in the short, high quantum energy range of F 660 2.88 – 4.9 5.18 – 5.6 0.290 TLLLT, direct photochemical activation is feasible, Luciferase Gene From American Firefly P. Pyrallis 690 2.70 – 4.0 1.44 – 2.4 0.03*energies at longer NIR wavelengths are typically 785 4.4 -4.2 0.64insufficient for the majority of photochemical •luciferin + ATP → luciferyl adenylate + PPi 808 3.62-1.2 0.84 PBS [5h] No-LLLT [5h] LLLT [5h]processes. However, other photophysical processes 905 CW 3.66 – 5.15 6.15 – 3.1 9.21-2.84 0.745 Semi-quantifiable differences in the macrophages and other invadingcan lead to activation of the immune system, even by •luciferyl adenylate + O2 → oxyluciferin + AMP + LIGHT cell within the synovial space are evident with more cells seen for 905 PW 15.45 – 5.1 6.61 – 4.1 0.021*short, transient, thermal effects. Monitoring immune reduced iNOS BLI signal and late time to maximum BLI counts.modulation effects due to LLLT can become a very Injection of Zymosan A: (yeast cell wall) in PBS P values considered statistically significant are representedpowerful method to understand underlying 12000 with asterisk (*).mechanisms. NO is a powerful modulator for the Laser irradiation: controlimmune system and inducible NO synthase (iNOs) a 10000 635 Discussion 15 min after injection BLI counts[photons/sec] 785direct surrogate for its activity. Exploiting Wavelengths: 635nm (n=5) 8000 808bioluminescence when the luciferase gene is placed 905 Laser therapy 15 minutes post- induction of inflammation 660nm (n=5) 6000 resulted in less recruitment of inflammatory cells to the site,within the promoter region of iNOs permits thetemporal monitoring of iNOs expression. In an acute 785nm (n=4) 4000 possibly by altering the mechanism of cell attachment by NOmodel of inflammation in the murine knee joint it 808nm (n=5) 2000was shown that a single treatment of 50mW/cm2 905nm (n=11) LLLT increased the expression of iNOS per cell with particularirradiance of a pulsed NIR light (905nm, 200nsec), 0 PW 905 also showing an earlier time to peak (~5 hrs) 0 5 10 15 20 25 30with 14mW power and 5J/cm2 radiant exposure, Irradiance: 50 mW cm-2 time[h](0.28cm2 spot size), low duty cycle, was most LLLT of 635nm leads to upregulation of iNOS expression, in radiant power: 5J cm-2 young animals but not as effectively as the longer wavelengthseffective in modulating iNOs expression, showing a iNOS related BLI signal as function of time post Zymosan Arapid onset. Histology showed an inverse correlation administration for a subset of mice. Average and stdev for n=4, young It seems that iNOS expression is playing a role in the anti- mice at , 15 weeks only. (from Moriyama Y. et. al. Photochem.of inflammatory cell concentration in the synovium Photobiol. 2005, 81:1351-1365 inflammatory mechanisms of LLLT.and integrated bioluminescence signal over the first Longer wavelength appear more effective in older animals.24hrs of inflammation induction. These resultssuggest that high iNOs expression may play a criticalrole in the anti-inflammatory mechanisms of LLLTexpression post inflammation may preventconversion of an acute inflammation into a chronicone. Luciferin injected and bioluminescence images LimitationsSee also: Moriyama, Y., Nguyen, J., Akens, M., acquired The study was executed over multiple years with various studentsMoriyama, E., Lilge, L. 2009. Lasers in Surgery and an technical staff performing the breading and genotyping and theMedicine. 41:227-231. experiments. Variability in determination of homozygous versus heterozygous state are small, however, success rates in Zymosan A and Luciferin administration can not be excluded. Unfortunately we could not randomize the treatment protocols over the Action spectra of LLLT on iNOS BLI signal. Solid symbols: animals <15 duration of these experiments. The number N of animals is still weeks. Open symbols: animals >15 weeks so younger than 8 months. low. Gray symbols: data from Ref. [13], with experiments executed in mice older than 1 year (signal corrected for homozygote vs. heterozygote used here). Triangles (~) refer to 905 nm in pulsed wave (PW) mode. Standard deviations are listed in Table I. Acknowledgement The authors with to express their gratitude to Drs. E. Moriyama, R. t=0h t=3h t=5h t=7h t=9h t=24h Weersink, and Miss J. Nguyen for their support of these studies. Funding was provided by Ontario Centres of Excellence, Photonics Research Ontario and Theralase Inc. 1