Vibrational spectroscopy of muscular tissue intoxicated by snake venom and exposed to photobiomodulation therapy
The pathogenesis of myonecrosis caused by myotoxins from bothropic venom is associated with local extracellular matrix (ECM) disintegration, hemorrhage, and inflammation. Search for alternative methods associated with serum therapy is mandatory to neutralize the fast development of local damage following snakebites. The experimental use of photobiomodulation therapy (PBMT) in murine models has shown promising results relative to structural and functional recovery from bothropic snakebite-induced myonecrosis. This study pioneered in using Raman and Fourier transform infrared (FTIR) spectroscopies to characterize biochemical alterations in the gastrocnemius that had been injected with Bothrops jararacussu venom and exposed to local PBMT. Results show that vibrational spectra from lyophilized and diluted venom (1307 cm −1) was also found in the envenomed gastrocnemius indicating venom presence in the unirradiated muscle 48 h post-injection; but any longer visible after PBMT at this time exposure or 72 h post-injection regardless irradiated or not. Raman and FTIR analyses indicated that the bands with higher area and intensity were 1657 and 1547 cm−1 and 1667 and 1452 cm−1, respectively; all are assignments for proteins, especially collagen, and are higher in the PBMT-exposed gastrocnemius. The infrared spectra suggest that laser treatment was able to change protein in tissue and that such change indicates collagen as the main target. We hypothesize that the findings reflect remodeling of ECM with key participation of collagen and faster tissue recovery for an anabolic condition.
KeywordsMyonecrosis PBMT Raman spectroscopy FTIR Snake venom Bothrops jararacussu
We are grateful to Professor Vitor Baranauskas (Brazilian Academy of Sciences, in memoriam) by his contribution to science in the last years and for all the support given for the realization of this work. Professor Baranauskas passed way in October, 2014.
This work was supported by grants from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) (Proc. 05/53625-1) (http://www.fapesp.br/) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, grant nos. 488792/2011 and 486142/2012-4) (http://www.cnpq.br/). MACH is a IA research fellow from CNPq (grant no. 305099/2011-6); WFV was a Master Sci. student granted with a scholarship from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) (http://www.capes.gov.br/) at the Department of Semiconductors, Instruments and Photonics (FEEC-UNICAMP) and Department of Biochemistry and Tissue Biology (IB-UNICAMP).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. The experiments were approved by the institutional Committee for Ethics in Animal Use (CEUA/UNICAMP, protocol no. 2950-1).
All authors have approved the manuscript and agree with submission to Lasers in Medical Science (LIMS).
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