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0The behavior of the protein-nanoparticle complex under laser-induced optical breakdown: an optical study
E. Nagaev1*, A. Vedunova2, T. Matveeva1, A. Simakin1, R. Sarimov1
1-Prokhorov General Physics Institute of the Russian Academy of Sciences, 119991 Moscow, Russia 2- Lobachevsky State University of Nizhny Novgorod, 603950 Nizhny Novgorod, Russia
* egorn97@mail.ru
Modern medicine operates state-of-the-art technologies. Among them laser surgery is on top. Surgical lasers made it possible to make minimal-invasive operations in comparison with traditional scalpels. Moreover, with surgical lasers it is possible to make operations, that could not be done before (for example, tissue welding). The results of the laser-tissue interaction depend on the parameters of the surgical lasers. Such lasers are divided by its operational wavelength (IR, visible, UV-region), working mode (continuous, pulsed) and beam power. Pulsed lasers are used in tissues welding, destruction of stones in kidneys, tattoo removal etc. It is well-known, that during laser-matter interaction non-linear effects can be occurred, when the pulse power is higher than the threshold. Among non-linear effects most studied are self-focusing, second and higher harmonic generation, laser-induced optical breakdown etc. It well-known, how laser-induced optical breakdown interacts with the tissues. But a little is known about such process in biological molecules solutions. Previously, we have shown how the typical surgical laser radiation interacts with BSA and HEWL solutions in-vitro [1,2]. It has been shown, that after 30 min of the experiment partial denaturation, aggregation and fragmentation of the protein molecules occurred. But the major volume of proteins remained as monomers. Modern nanotechnologies made it possible to use synthetic nanoparticles in bioimaging and therapy. This technological stack, when nanoparticles are used in the imaging and therapy, is called theranostics. Thus, there is a question: what would be happened, when the operation with the use of surgical laser would be in the presence of such nanoparticles?
In this work, we used laser-synthesized gold nanoparticles and the HEWL solutions with protein concentrations of 1 mg/ml and 8 mg/ml. Gold nanoparticles were added to protein solutions with the final concentration of nanoparticles of 1011 NPs/ml. The resulting solutions were irradiated with the Nd:YAG laser with second harmonic generation at 532 nm. The longest exposure time was 30 min. This time was chosen because some surgical operations are more than 30 min long. It is shown, that after adding gold nanoparticles to protein HEWL solutions, protein corona was formed. After exposure optical characteristics of the solutions were recorded. It shown, that the absorbance region, associated with nanoparticles surface plasmon resonance, changed over time - plasmon resonance peak decreased and longer-wavelength absorbance have been increased. Dynamic light scattering distributions shown, that the peak, associated with the protein monomers, has been disappeared after adding of NPs. After 30 min exposure this peak has been appeared, as in the control samples. Emission-excitation matrixes shown, that the amino acid peaks have been decreased (in the case of the protein concentration 8 mg/ml more than 50% decrease). Refraction indexes of both solutions have been increased, that can be associated with the partial denaturation of HEWL molecules. Activity of the protein solutions in the presence of the gold NPs, decreased. But the major volume of protein molecules stayed in the form of monomers.
Funding: This work was supported by a grant of the Ministry of Science and Higher Education of the Russian Federation (075-15-2022-315) for the organization and development of a World-class research center "Photonics".
[1] E.I. Nagaev, I.V. Baimler, A.S. Baryshev, M.E. Astashev, S.V. Gudkov, Effect of laser-induced optical breakdown on the structure of BSA molecules in aqueous solutions: An optical study, Molecules, 27(19), 6752, (2022).
[2] R.M. Sarimov, T.A. Matveyeva, V.A. Mozhaeva, A.I. Kuleshova, A.A. Ignatova, A.V. Simakin, Optical Study of Lysozyme Molecules in Aqueous Solutions after Exposure to Laser-Induced Breakdown, Biomolecules, 12(11), 1613, (2022).
