S1

SUPPORTING INFORMATION (SI)

The rational surface design of upconversion nanoparticles with
polyethylenimine (PEI) for biomedical applications: better safe than brighter?
Anna E. Guller,†,‡,§,◆,∇ Annemarie Nadort,*,†,∇ Alla N. Generalova,∥,¶,∇ Evgeny V. Khaydukov,‡,¶ Andrey V.
Nechaev,⊥ Inna A. Kornienko,# Elena V. Petersen,# Liuen Liang,† Anatoly B. Shekhter,‡ Yi Qian, § Ewa
M. Goldys,†,◆ and Andrei V. Zvyagin†,‡,○

†
Macquarie University, Department of Physics and Astronomy, MQ Photonics Research Centre, The ARC
Centre of Excellence for Nanoscale BioPhotonics, North Ryde, New South Wales 2109, Australia
‡
Sechenov First Moscow State Medical University, Moscow, 119991, Russia
§
Macquarie University, Department of Biomedical Sciences, North Ryde, New South Wales 2109,
Australia
∥Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry of the RAS, Moscow, 117997, Russia

⊥Institute of Fine Chemical Technologies, Moscow Technological University, Moscow, 119571, Russia
#
Moscow Institute of Physics and Technology, Dolgoprudnyi, 141700, Russia
¶
Scientific Research Centre “Crystallography and Photonics”, Russian Academy of Sciences, Moscow,
119333, Russia
○
Lobachevsky Nizhniy Novgorod State University, Nizhny Novgorod, 603022, Russia
◆University of New South Wales, Sydney, New South Wales 2032, Australia

∇ Equally contributing authors

*Corresponding author: Annemarie Nadort, PhD, Research Fellow,
E-mail: annemarie.nadort@mq.edu.au

Number of pages; 5
Figures: Figure S1 and S2 (ICP-MS validation study), and Figure S3 (confocal images)
Tables: Table S1 (ESI 2. Detailed statistical analysis viability tests)
 S2

SI 1. ICP- MS validation study

ICP-MS linear response for UCNP colloids

4.E+08
R² = 1
4.E+08
3.E+08
Yttrium counts

3.E+08
2.E+08
2.E+08
1.E+08
5.E+07
0.E+00
0 20 40 60 80 100 120
Relative colloid concentration (%)

Figure S1. We confirmed that the ICP-MS technique measures an Y-ion content count that is linear with the particle
content in a colloid solution. We serially diluted the colloidal UCNPs in 2% HNO 3 without further digestion and
recorded the ICP-MS Y-counts, which shows an excellent linearity over more than 3 orders of magnitude dilution of
the colloid (r2 > 0.99).

ICP-MS repeatability for UCNP colloids

5
% distance from mean value

4
3
2
1
0
-1
-2
-3
-4
-5
0 2 4 6 8
measurement repeat

Figure S2. Repeatability of ICP-MS technique for Y-content of colloidal solutions of UCNPs. The measurements
were taken at 2 different days and show high repeatability; all Y-counts are within 5% of the mean sample value,
with an average deviation of ±1.4 % from the mean value.
 S3

SI 2. Detailed statistical analysis viability tests

Table S1. ANOVA Post Hoc Tests for multiple comparisons (Bonferroni) of viability of keratinocytes, %.
Exposure 95% Confidence
time, h Mean Difference Std. Interval
(I) Type of particles (J) Type of particles Sig.
(I-J) Error Lower Upper
Bound Bound
24 TMAH-UCNP 23.97(*) 3.94 .00 10.90 37.05
UCNP@PEI 46.83(*) 3.75 .00 34.40 59.25
UCNP@PEI@DxS 27.54(*) 3.75 .00 15.11 39.97
UCNP@PEI@Dx 50.59(*) 3.75 .00 38.16 63.02
Control
UCNP@PEI@PAMA 25.60(*) 3.75 .00 13.17 38.03
UCNP@PEI@PAMA@Ch 33.19(*) 3.75 .00 20.76 45.62
UCNP@PEI@PAMA@Dx 33.29(*) 3.75 .00 20.86 45.72
UCNP@PEI@SCh 22.65(*) 3.75 .00 10.22 35.08
UCNP@PEI 22.85(*) 4.75 .00 7.10 38.61
UCNP@PEI@DxS 3.57 4.75 1.00 -12.19 19.32
UCNP@PEI@Dx 26.62(*) 4.75 .00 10.86 42.37
TMAH-UCNP UCNP@PEI@PAMA 1.63 4.75 1.00 -14.13 17.38
UCNP@PEI@PAMA@Ch 9.22 4.75 1.00 -6.54 24.97
UCNP@PEI@PAMA@Dx 9.32 4.75 1.00 -6.44 25.07
UCNP@PEI@SCh -1.32 4.75 1.00 -17.08 14.43
UCNP@PEI@DxS -19.29(*) 4.59 .00 -34.51 -4.07
UCNP@PEI@Dx 3.76 4.59 1.00 -11.46 18.98
UCNP@PEI@PAMA -21.23(*) 4.59 .00 -36.44 -6.01
UCNP@PEI
UCNP@PEI@PAMA@Ch -13.64 4.59 .14 -28.86 1.58
UCNP@PEI@PAMA@Dx -13.54 4.59 .15 -28.76 1.68
UCNP@PEI@SCh -24.18(*) 4.59 .00 -39.39 -8.96
UCNP@PEI@Dx 23.05(*) 4.59 .00 7.83 38.27
UCNP@PEI@PAMA -1.94 4.59 1.00 -17.16 13.28
UCNP@PEI@DxS UCNP@PEI@PAMA@Ch 5.65 4.59 1.00 -9.57 20.87
UCNP@PEI@PAMA@Dx 5.75 4.59 1.00 -9.47 20.97
UCNP@PEI@SCh -4.89 4.59 1.00 -20.11 10.33
UCNP@PEI@PAMA -24.99(*) 4.59 .00 -40.21 -9.77
UCNP@PEI@PAMA@Ch -17.40(*) 4.59 .01 -32.62 -2.18
UCNP@PEI@Dx
UCNP@PEI@PAMA@Dx -17.30(*) 4.59 .01 -32.52 -2.08
UCNP@PEI@SCh -27.94(*) 4.59 .00 -43.16 -12.72
UCNP@PEI@PAMA@Ch 7.59 4.59 1.00 -7.63 22.81
UCNP@PEI@PAMA UCNP@PEI@PAMA@Dx 7.69 4.59 1.00 -7.53 22.91
UCNP@PEI@SCh -2.95 4.59 1.00 -18.17 12.27
UCNP@PEI@PAMA@Dx .10 4.59 1.00 -15.12 15.32
UCNP@PEI@PAMA@Ch
UCNP@PEI@SCh -10.54 4.59 .88 -25.76 4.68
UCNP@PEI@PAMA@Dx UCNP@PEI@SCh -10.64 4.59 .83 -25.86 4.58
120 TMAH-UCNP 28.92(*) 6.86 .00 6.14 51.71
UCNP@PEI 86.94(*) 6.86 .00 64.15 109.73
UCNP@PEI@DxS 42.21(*) 6.86 .00 19.42 65.00
Control UCNP@PEI@Dx 84.97(*) 6.86 .00 62.19 107.76
UCNP@PEI@PAMA 47.82(*) 6.86 .00 25.04 70.61
UCNP@PEI@PAMA@Ch 38.81(*) 6.86 .00 16.02 61.60
UCNP@PEI@PAMA@Dx 45.21(*) 6.86 .00 22.42 68.00
 S4

UCNP@PEI@SCh 39.17(*) 6.86 .00 16.39 61.96

UCNP@PEI 58.01(*) 8.20 .00 30.77 85.25
UCNP@PEI@DxS 13.29 8.20 1.00 -13.95 40.53
UCNP@PEI@Dx 56.05(*) 8.20 .00 28.81 83.29
TMAH-UCNP UCNP@PEI@PAMA 18.90 8.20 .86 -8.34 46.14
UCNP@PEI@PAMA@Ch 9.89 8.20 1.00 -17.35 37.13
UCNP@PEI@PAMA@Dx 16.29 8.20 1.00 -10.95 43.53
UCNP@PEI@SCh 10.25 8.20 1.00 -16.99 37.49
UCNP@PEI@DxS -44.73(*) 8.20 .00 -71.96 -17.49
UCNP@PEI@Dx -1.96 8.20 1.00 -29.20 25.28
UCNP@PEI@PAMA -39.11(*) 8.20 .00 -66.35 -11.87
UCNP@PEI
UCNP@PEI@PAMA@Ch -48.13(*) 8.20 .00 -75.36 -20.89
UCNP@PEI@PAMA@Dx -41.73(*) 8.20 .00 -68.96 -14.49
UCNP@PEI@SCh -47.76(*) 8.20 .00 -75.00 -20.52
UCNP@PEI@Dx 42.76(*) 8.20 .00 15.52 70.00
UCNP@PEI@PAMA 5.61 8.20 1.00 -21.63 32.85
UCNP@PEI@DxS UCNP@PEI@PAMA@Ch -3.40 8.20 1.00 -30.64 23.84
UCNP@PEI@PAMA@Dx 3.00 8.20 1.00 -24.24 30.24
UCNP@PEI@SCh -3.04 8.20 1.00 -30.28 24.20
UCNP@PEI@PAMA -37.15(*) 8.20 .00 -64.39 -9.91
UCNP@PEI@PAMA@Ch -46.16(*) 8.20 .00 -73.40 -18.92
UCNP@PEI@Dx
UCNP@PEI@PAMA@Dx -39.76(*) 8.20 .00 -67.00 -12.52
UCNP@PEI@SCh -45.80(*) 8.20 .00 -73.04 -18.56
UCNP@PEI@PAMA@Ch -9.01 8.20 1.00 -36.25 18.23
UCNP@PEI@PAMA UCNP@PEI@PAMA@Dx -2.61 8.20 1.00 -29.85 24.63
UCNP@PEI@SCh -8.65 8.20 1.00 -35.89 18.59
UCNP@PEI@PAMA@Dx 6.40 8.20 1.00 -20.84 33.64
UCNP@PEI@PAMA@Ch
UCNP@PEI@SCh .36 8.20 1.00 -26.88 27.60
UCNP@PEI@PAMA@Dx UCNP@PEI@SCh -6.04 8.20 1.00 -33.28 21.20

* The mean difference is significant at the .05 level.

 S5

S2 Confocal imaging of HaCaT keratinocytes incubated with UCNPs

Figure S3. Confocal images showing the uptake of UCNP@PEI by HaCaT keratinocytes after 48 h exposure
in vitro. The images consist of an overlay of bright field and UCL (green colour) images. It can clearly be seen
that the optimal focus of the UCNPs (smallest spot) coincides with the optimal focus of the cells (Z = 6 um),
thus confirming that the UCNPs are located within the cell and not above or below.