Sigma Xi Student Research Competition Presentation


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Sigma Xi Student Research Competition Presentation

  1. 1. Effect of the Nano-Bio Interface on theGenotoxicity of Titanium Dioxide Nanoparticles and Associated Cellular Response R u Y. Pr asad aj Ph.DC andi dat e Envi r onm al Sci ence and Engi neer i ng D ent epar t ment U C G l l i ngs’ G obal School of Publ i c H t h N i l eal GPSF U ver si t y R ni esear ch Day Febr uar y 26, 2012 1
  2. 2. Brief History of Engineered Nanoparticles“There’s Plenty of Room at the Bottom” – R char d Feynm N i an, obel Laur eat ePhysi ci st , 1959 1985 – Fi r st 2001 – Pr esi dent Devel opm of ent nanopar t i cl e C i nt on est abl i shes l Nanopar t i cl es f or :synt hesi zed (car bon N i onal at f ul l er enes) Nanot echnol ogy I ni t i at i ve • D ug D i ver y r el • C onsum Pr oduct s er • I ndust r i al Pr oduct s “The 2013 Federal Budget provides $1.8 billion for the National Nanotechnology Initiative (NNI)” 2
  3. 3. Definition of Engineered Nanoparticles ~1.6 m ~2 nm width 100 10-1 10-2 10-3 10-6 10-9 1-100 nm meter decimeter centimeter millimeter micrometer nanometer Par t i cl es w t h a si ze di m i ensi on bet ween 1 and 100nmt hat shownovel pr oper t i es t hat ar e not f ound i n bul k sam es of t he sam m er i al pl e at 3
  4. 4. Titanium Dioxide Nanoparticles C yst al St r uct ur es: Anat ase, R i l e and r ut Br ooki t eH gh R r act i ve I ndex, W t e Pi gm at i on i ef hi ent 4
  5. 5. Use of Titanium Dioxide Nanoparticles in Consumer Products Sunscreens : ~10-100 µg Ti/mg product (1-10%) Weir et al. Environ Sci Technol 2012 5
  6. 6. Association of Physicochemical Properties with ToxicityD t o sm l si ze di m ue al ensi ons, l ar ge sur f ace ar ea t o vol um r at i o, novel pr oper t i es, and pot ent i al e exposur e, i t i s i m t ant t o det er m ne ef f ect s on por i hum heal t h an Sayes et al. Tox Sci 2006 Ther e i s a need t o det er m ne physi cochem cal i i char act er i st i cs of nanopar t i cl es t hat can l ead t o adver se 6 ef f ect s
  7. 7. Nano-Bio InterfaceFor m i on of Pr ot ei n at I nt er act i on w t h C l s i elC ona or Nel et al. Nature Materials 2009 7
  8. 8. Impact of Nano-Bio Interface on Genotoxicity Degussa P25 Ti O 2 nanopar t i cl es el ul edi :C l C t ur e M um -fSer um r ee K M G KB DM KF • C l C t ur e m um el ul edi • PBS + 0.6% BSA + 0.01% • C l C t ur e m um el ul edi (KG ) + 0.1% BSA M DSPC (sur f act ant ) (KG ) + 10% FBS M • 2 m n pr obe i • 20 m n cup soni cat i on i • 2 m n pr obe i soni cat i on of st ock of st ock sol ut i on soni cat i on of st ock sol ut i on • C r i f uged and r e- ent sol ut i on suspended i n cel l cul t ur e m um(KG ) edi M Ef f ect s on H an Lung Epi t hel i al C l s (BEAS- um elPrasad et al., 2013 ACS 2B) Nano 8
  9. 9. Agglomeration: Physicochemical Characteristic Impacted by the Nano-Bio Interface
  10. 10. Study Hypothesis The t r eat m m umt hat el i ci t s t he ent edism l est nanopar t i cl e aggl om at es w l l al er ibe associ at ed w t h cel l ul ar i nt er act i on i and genot oxi ci t y
  11. 11. Identification of the Protein Adsorbed to the Surface of the Nanoparticles KB DM KF 2 Ti O i n K 2 2 B Ti O i n D Ti O i n K M F ladder kDa 250 130 95 72 55 36 28Tedja et al., 2012 ACS Nano, Lundqvist et al., 2008 PNAS 11
  12. 12. Size Measurements in Treatment Media0h KB24 h DM KFPrasad et al., 2013 ACS Nano 12
  13. 13. No Significant Cytotoxicity in BEAS-2B cells 0 µg/ml 10 µg/ml 20 µg/ml 50 µg/ml 100 µg/ml MMS (100 µM) Li ve: C cei n- al AM D ead: Pr opi di um I odi dePrasad et al., 2013 ACS Nano 13
  14. 14. Highest Cellular Uptake in KF Medium Determined by Flow Cytometry Fl owC om r y H st ogr am yt et i sPrasad et al., 2013 ACS Nano 14
  15. 15. Evaluation of Genotoxicity (Damage to the Genome) D A D age N am C om hr osom D age al am Binucleated cell (BN) treat 24 h 48 h Add CytoSpin cells cytochalasin B on glass slides Micronucleated Binucleated cell (MN)Fenech et al. Mutat Res-Fund Mol M 2000 , Kirsch-Volders et al. Mutat Res-Fund Mol M 2003, Singh et al. Mutat Res 1991 15
  16. 16. TiO2 Nanoparticles Induce DNA Damage C r ol ont D aged amPrasad et al., 2013 ACS Nano 16
  17. 17. Micronuclei Induction Only in KF MediumPrasad et al., 2013 ACS Nano 17
  18. 18. Increased % of S-Phase Cells in KF Medium C l C e el yclPrasad et al., 2013 ACS Nano 18
  19. 19. Summary M umt hat el i ci t ed edi sm l est aggl om at es (K al er F) w associ at ed w t h hi ghest as i cel l ul ar upt ake, m cr onucl ei i f or m i on, and i ncr eased % of at cel l s i n S phase. I n cont r ast , D A dam w N age as i nduced r egar dl ess of aggl om at i on and cel l ul ar er upt ake I nhal at i on exposur e t o t i t ani umdi oxi de nanopar t i cl es m not i nduce ay m cr onucl ei i n l ung i cel l s, whereas, serum proteins in “blood” (f or exam e, i ngest i on exposur e) plPrasad et al., 2013 ACS Nano m act t o l ow ay er 19
  20. 20. Thank youDr. Rebecca Fry Dr. David DeMarini Dr. Ivan Rusyn Dr. Jim Swenberg Dr. Bill Kaufmann Kathleen Wallace Dr. Carl Blackman Dr. Nana Nikolaishvili-Feinberg Kaitlin Daniel Dr. Jorge Muniz Ortiz Dr. Stephanie Cohen Alan Tennant Dr. Andrew Kligerman Mark Olorvida Jim Campbell Dr. Robert Zucker Bentley Midkiff Micaela Killius Dr. Steve Simmons Mervi Eeva Sharon Leavitt Dr. Kevin Dreher Dr. Stephanie Smith-Roe Ben Castellon Dr. Bellina Veronesi Lisa Smeester Jenna Strickland Dr. Brian Chorley Dr. Kathryn Bailey Danielle Ackerman Dr. Thomas Knudsen Dr. Bhavesh Ahir Peggy Matthews Dr. Kirk Kitchin Dr. Julia Rager Sarah Warren Dr. Will Boyes Alison Sanders Nancy Hanley Dr. Bill Lefew Fry lab Laura Degn Keith Tarpley Kristen Sanders U.S. EPA NHEERL Questions?