Magnetic-modified nanodiamond for bio-applications

Transcription

1 Magnetic-modified nanodiamond for bio-applications Chang-You Song 1, Nikolai Perov 2, Valentina Bessalova (Samsonova) 2, Svetlana Norina 2, Li-Chi Liu 1, Zhe-Rui Lin 1, Yu-Chung Lin 1, Ashek-I-Ahmed 1, Artashes Karmenyan 1, Olga Levinson 3, Boris Zousman 3, Elena Perevedentseva 1, Chia-Liang Cheng 1,* 1 Department of Physics, National Dong Hwa University, Taiwan 2 Moscow State University, Russia 3 Ray Techniques Ltd., Israel NDNC 2016, Xi an China, May 22-26,

2 Nanodiamond use for bioapplications ND Carboxylation Functionalization - with linker Protein- Growth Hormone Protein- Lysozyme Protein- X ND-X complex AcO OH O Ph NH O O O + Ph O H OH OH OAc OCOPh Taxol Drug O O Carboxylation modifies ND surface with carboxyl groups Carboxylated ND can be functionalized with biomolecules Drug delivery, delivery tracing, targeted interaction, etc. 2

3 Nanodiamond use for bioapplications RBC Cancer cell Bacteria Microorganisms We studied ND interaction with different bio-systems and have found: Nontoxic / bio-compatible Great property of photoluminescence High efficiency in drug delivery Zebrafish embryo Mice Combine the properties of ND with magnetism. Applications of NMR, MRI, controlled drug delivery, 3 Nanomedicine 2013, 8,

4 The Advantages of Magnetic NPs for Bioapplications (1) Drug reservoir (2) Active accumulation H. Hao et al, J. Mater. Chem. B, 2014, 2, (3) Magnetic Resonance Image 4 L. H. Reddy, et. al., Chem. Rev., 2012, 112 (11),

5 M (emu/g) The Magnetic ND (RayND-M) ND powder was treated chemically and thermally in medium free of metals by the method developed by Ray Techniques Ltd. [J. Nanoscience & Nanotechnology, 15(2), (2015)] DND RayND-M H (Oe) It was found that this specific treatment of ND surface resulted in strong magnetism (ferromagnetism) of diamond nanoparticles. Studies of magnetism in carbon systems see review paper of T. Makarova 2004, Semiconductors 38, Energy Dispersive Spectrum RayND-M on Si wafer Magnetic atoms admixtures are negligible: Fe: weight%, Ni: 0 weight%; Co: weight%

6 Intensity (a.u.) Intensity (a.u.) Number (%) Characterization of Magnetic ND (RayND-M): size, surface, structure O-H C=O C-H Ray ND-M cnd Average: 90 nm wavenumber (cm -1 ) FTIR spectrum: The COOH can be observed in RayND-M cm cm -1 sp3 sp2 ex : 488 nm Size (nm) Size Distribution: Size: 90 nm. SEM image: High aggregation of crystallites is observed. z-potential: mv at ph Raman shift (cm -1 ) Raman spectrum: The shift of 1332 cm -1 to 1324 cm -1 shows that size of crystallites should be small.

7 Intensity (a.u.) Intensity (a.u.) Characterization of Magnetic ND (RayND-M): photoluminescence sp3 sp2 Diamond 569 nm RayND-M 5 nm cnd wavelength (nm) ex : 488 nm 641 nm The comparison of photoluminescence of Ray ND-M with 5 nm cnd. It shows the Diamond signal and ZPL with 488 excitation ex : 532 nm wavelength (nm) 7

8 Intensity (a.u.) Number (10 5 ) Characterization of Magnetic ND (RayND-M): 2-photon excited fluorescence Fluorescence 2p-lifetime imaging of RayND-M on glass substrate Time decay (ns) Lifetime decay of RayND- M luminescence Time (ns) Histogram of lifetimes distribution We observe enough bright 2-photon excited signal, allows 2-photon imaging, in perspective - lifetime and time-gate imaging 2-photon excitation in red and infrared range is done with femtosecond tunable Ti-Sapphire laser (760 nm, 80 MHz, 140 fs)

9 9 cell viability (%) cell viability (%) Interaction with cells: Cytotoxicity in BHK (Baby hamster kidney) cell RayND-M hr 100 * * ** ** 50 0 control g/ml RayND-M + HSA hr 100 * 50 0 control g/ml Some concentration-dependent cytotoxicity is observed The toxicity can be significantly decreased by coating of ND particles surface with albumin (Human Serum Albumin)

10 Interaction with cells: Fluorescence confocal imaging Z-scan: Confocal image of BHK cell after interaction with ND for 24 hrs. As Z-scan demonstrates: ND penetrates into cell (and also some ND is attached on the cell membrane). ND is well detectable in cell. Excitation Emission RayND 514 nm nm Nuclei 405 nm nm Cytoplasm 633 nm nm

11 Intensity (a.u.) number (10 4 ) Interaction with cells: 2-photon excitation and fluorescence lifetime imaging Very good fluorescence signal of RayND-M at 2-photon excitation with femtosecond tunable Ti-Sapphire laser (760 nm, 80 MHz, 140 fs) Makes this ND promising for 2-photon imaging and lifetime imaging : Control without ND autofluorescence of BHK cell BHK cell with RayND-M Time delay (ns) cell autofluorescence RayND_M Lifetime decays allow distinguish ND and cell signal Time (ns) Cell with ND M-ND Cell Histograms of lifetime distribution

12 Summary Strong magnetism of nanodiamond is observed and the nanodiamond is characterized for it s application in bio-medical studies. The surface and luminescence properties give the possibilities for biomedical applications. In the applications magnetic and photoluminescence properties can be combined. The nature of magnetism is unclear yet and under study. 12

13 Acknowledgements: To MOST, Ministry of Science and Technology of Taiwan Thank you for your attention! 13

14 Zeta potential (mv) Absorption ph Wavelength (nm) Tools 1. LakeShore 7407 vibrating sample magnetometer (VSM) (Lake Shore Cryotronics, US), 2. Renishaw, UK, 3. - SNOM, Witec, Germany, 4. Jobin Yvon, T64000, France-Japan 5. Scanning Fluorescence Confocal Microscope (TCS-SP5, Leica, Germany) 6. System for lifetime imaging on the base of femtosecond tunable Ti-Sapphire laser (760 nm, 80 MHz, 140 fs) and PicoQuant scanner Etc 14

15 Magnetic properties of carbons and nanocarbons are recently studied. Pure diamond structure combines diamagnetic and paramagnetic compounds. M, arb. unit But in an intermediate graphite diamond structure spin ordering and magnetic interactions can exist [T. L. Makarova Semiconductors, 2004, 38, 6, ] Ferromagnetic properties of carbon nanostructures are discussed for theoretically predicted structures and are shown experimentally. Particularly, it has been predicted that structures composed of altered C atoms with different hybridization (sp 2 - and sp3 coordinated) can have strong spontaneous magnet moment /under some conditions strong regular ordering difficult to realize, but on the micro-level such combination is realized in ND/ Superparamagnetic or/and ferromagnetic particles Ferromagnetic: 15