CC BY
15B-I-16
BIOMEDICAL PHOTONICS
Effect of various NP on in vitro development of preimplantation
mouse embryos
A. Karmenyan1, A. Krivokharchenko2, M. Kormacheva1, M. Sarmiento1, E. Barus1,
E. Perevedentseva3, C.-L. Cheng1
1-Department of Physics, National Dong Hwa University, Hualien 97401, Taiwan 2 - N.N. Semenov Institute of Chemical Physics, Moscow 119991, Russia 3- P. N. Lebedev Physics Institute, Moscow, 119991, Russia artashes@gms.ndhu.edu.tw
Recently, with the tremendous progress in nanotechnologies, the use of various types of nanoparticles (NPs) in mass production and in number of applications has become widespread. Among other applications NPs due to their unique physicochemical properties are promising for biomedical use [1] as a drug carriers, biosensors, coating for medical implants and more. Various surface functionalizations are developed to customize nanomaterials for different application. Despite all the advantages of NPs, there is an enormous variety of cells, the biological effect on which is still insufficiently studied and unpredictable. Thus, development of reproductive technologies (for human and animals) including in-vitro manipulation with early embryos, can lead to increase the interaction probability between NP and the preimplantation embryos.
In this work, we are presenting an express examination of the effect of few different types of NPs on the development of a preimplantation embryo during prolonged incubation (in some cases, during the entire period of development until hatching from the zone pelucida (ZP)), the interaction of NPs with the ZP of oocytes and early mammalian embryos. The assessment of the embryo state included the observation of morphological changes, the period of development, the possibility of passing into the zone, and inside the developing embryo.
The 2-cell stage mice embryos and the oocytes have been used for observation interaction with different kinds of nanodiamonds (ND), titanium dioxide (TiO2), graphene oxide (GO) particles. The NPs tested were selected due to their wide varied application in many household products, technological and industrial processes and materials; - rapidly growing interest in bio-medical research and applications. They also have previously demonstrated non-toxicity for cellular models and applicability for bioimaging applications and some bio-optical therapies [2,3]. Physical characteristics of used NPs were analyzed (UV-VIS absorption, FTIR and Raman spectra, size distributions, and z-potential were measured). Their interaction with embryos and oocytes was observed using methods of confocal laser microscopy, Raman spectroscopy and fluorescence lifetime imaging with two-photon excitation (TP-FLIM) and analyzed in correspondance with the development observations.
No strong disturbing effect of NPs in applied concentrations neither well-observable penetration of the NDs and TiO2 into the embryos has been detected. Note that it can opens more opportunities for controllable nanobioapplications of these NPs. Additionally, using TP-FLIM gave the possibilities to detect the penetration of GO NPs through the ZP and to discuss the possible mechanisms of the NP penetration into the ZP and through the ZP into embryos and of relationship between the NPs surface properties, spectral characteristics, and their interaction with the ZP.
Acknowlegement: This work partly was supported by the grant MOST 109-2923-M-259-001-MY3
[1] K. Riehemann, S.W. Schneider, T.A. Luger, B. Godin, M. Ferrari, and H. Fuchs, Nanomedicine—Challenge and Perspectives, Angew. Chem. Int. Ed., 48, pp. 872-897, (2009).
[2] E. Perevedentseva, Y.-C. Lin, Mona Jani, and C.-L. Cheng, Biomedical applications of nanodiamonds in imaging and therapy, Nanomedicine, 8, pp. 2041-2060, (2013).
[3] E. Perevedentseva, D. Peer, V Uvarov, B. Zousman, and O. Levinson, Nanodiamonds of laser synthesis for biomedical applications, J. Nanoscience & Nanotechnology, 15, pp. 1045-1052, (2015).
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