CHAPTER 4: SUPPORTING INFORMATION
kDa 250 130 95 72 55 36 28 Ladder 1 2 3 4
Supplemental Figure 11. Proteins adsorbed to the surface of nano-TiO2 in DB or DF medium after 24 h as visualized by 1D SDS-PAGE gel electrophoresis. Nano-TiO2 were dispersed in treatment medium and incubated for 24 h. After incubation, nano-TiO2 were centrifuged and washed with PBS. The PBS was aspirated and nano-TiO2 was mixed with SDS-PAGE buffer for 10 minutes at 95°C to allow for adsorbed proteins on the surface of nano-TiO2 bind to buffer. Ladder) Fermentas PageRuler™ protein ladder, (1) nano-TiO2 in DB medium, (2) DB medium alone, (3) nano-TiO2 in DF medium, (4) DF medium alone.
Supplemental Figure 12. Cell viability was evaluated by Cell-Titer Blue® after exposure to nanoparticles at 0.003 – 100 µg/ml in half log increments. A 24-h exposure to silver nitrate (AgNO3) at 0.3 to 100 µg/ml in half log increments was used as a positive control. *p < 0.05, n = 3. * * * * * * DB# DF#
148 F ol d c h an ge fr om u n tr eate d c on tr ol F ol d c h an ge fr om u n tr eate d c on tr ol
log [material] (µg/ml) log [material] (µg/ml)
Supplemental Figure 13. Fold changes in expression of cellular stress-response luciferase-reporter genes for (A) NFkB and (B) AP1 exposed to the positive control hydroquinone (HQ) for 24 h in HepG2 cells at half log concentrations. *p < 0.05, n = 3.
149
Characteristic ```````` Result
Primary Particle Size 27.5 nm
Size Range 14.6-64.1 nm
Surface Area 49 m2/g
% Purity 95.1%
Crystal Form 86% anatase/14%rutile
Supplemental Table 5. Physical characteristics of nano-TiO2a
aSize, surface area, purity, and crystal form of P25 TiO
2 received
from manufacturer were measured independently. Approximately 150 dry particles were examined by TEM. Data from the University of Kentucky (see Materials and Methods).
Supplemental Table 6. Polydispersion index and zeta potential of nano-TiO2 in KB and KFa
Treatment Medium
Concentration (µg/ml)
Polydispersion Index Zeta Potential (mV)
DB
0
0.18 ± 0.03
-5.42 ± 1.90
10
0.31 ± 0.02
-8.11 ± 2.18
50
0.31 ± 0.08
-8.64 ± 0.84
100
0.26 ± 0.05
-7.22 ± 1.20
DF
0
0.18 ± 0.05
-6.48 ± 1.32
10
0.34 ± 0.05
-6.66 ± 0.48
50
0.32 ± 0.05
-8.48 ± 2.08
100
0.26 ± 0.07
-7.53 ± 0.82
Concentration (µg/ml)
NP
0
1
3
DB
4270 ± 1220 3957 ± 853 4297 ± 733
DF
3816 ± 405 3945 ± 479 3331 ± 528
Supplemental Table 7. Average number of Nuclei/WellaaData expressed as mean ± SD of three independent experiments.
KB
KB-Plus
KGM medium
KGM medium
1.08 g/L glucose
4.50 g/L glucose
1.50 mM KCl
5.37 mM KCl
Supplemental Table 8. KB and KB-Plus medium
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