Supplementary Materials for
Autonomous metal-organic framework nanorobots for active mitochondria-
targeted cancer therapy
Xiqi
Peng
et al
.
Corresponding author: Songsong Tang, sstang@caltech.edu; Wei Gao, weigao@caltech.edu;
Song Wu, wusong@szu.edu.cn
Sci. Adv.
9
, eadh1736 (2023)
DOI: 10.1126/sciadv.adh1736
The PDF file includes:
Figs. S1 to S20
Legends for movies S1 to S7
Other Supplementary Material for this manuscript includes the following:
Movies S1 to S7
F
ig. S1. TEM images of ZIF
-
67 NPs (A
-
B) and ZIF
-
67@DOX
-
TPP nanorobots (C
-
D).
Scale
bars: 1 μm (
A,C
); 50 nm (
B,D
).
Fig. S2. Hydrodynamic size distribution of ZIF
-
67@DOX
-
TPP nanorobots.
F
ig. S3. Characterization of ZIF
-
67@DOX
-
TPP nanorobots and
their counterparts.
(
A
)
Wide scan of XPS spectra. Co 2p spectra of (
B
) ZIF
-67@DOX-
TPP nanorobots, (
C
) ZIF
-
67@DOX, and (
D
) ZIF
-
67 NPs. (
E
) XRD spectra and (
F
) UV
-vis absorption of ZIF
-67 NPs, ZIF
-
67@DOX, and ZIF-
67@DOX-TPP nanorobots.
F
ig. S4 .
Fluorescent
images of ZIF
-
67@DOX
-
TPP nanorobots.
Scale bar, 2 μm
.
Fi
g. S5.
Propulsion analysis of ZIF
-
67 NPs and ZIF
-
67@DOX
-
TPP nanorobots in PBS
solution with 100 μM H
2
O
2
.
(
A
) Typical motion trajectories (over 20 s). (
B
) mean square
displacement (MSD)
(n=15; mean ± SEM).
F
ig. S6. Propulsion analysis of ZIF
-
67@DOX
-
TPP nanorobots in 1640 culture medium with
various H
2
O
2
concentrations.
(
A
) Typical motion trajectories (over 20 s). (
B
) MSD. (
C
) D
eff
. (
D
)
Speed (n = 15; mean ± SEM).
F
ig. S7. TEM images of ZIF
-
67@DOX
-
TPP nanorobots after incubation in PBS solution
with different pH and H
2
O
2
concentrations for 12, 24 and 48 hours.
Scale bar, 200 nm.
F
ig. S8.
Drug loading and release profiles of ZIF
-
67@DOX NPs.
Drug loading capacity of ZIF
-
67@DOX NPs upon various incubation time (
A
) and DOX input concentrations (
B
) (n = 3; mean
± SD). The cumulative release of DOX from NPs upon different pH values (
C
) and H
2
O
2
concentrations (
D
) (n = 3; mean ± SD).
F
ig. S9. Cytotoxicity of ZIF
-
67 NPs to different cell lines.
(
A
) T24 bladder cancer cells. (
B
)
BEND3 brain endothelial cells. (
C
) SV
-
HUC
-
1 urothelial cells (n = 3; mean ± SD).
F
ig. S10 . The co
-
localization of ZIF
-
67@DOX
-
TPP nanorobots and lysosomes in T24
bladder cancer cells after short and long incubation durations.
The nuclei and lysosome were
stained with Hoechst 33342 (blue) and LysoTracker Green dye, respectively.
The right columns
show the calculated PCC of the colocalization of lysosome and nanorobo
ts in T24 cells (n = 5;
mean ± SD).
Scale bars, 10 μm.
F
ig. S11.
Morphology characterization and propulsion analysis of ZIF
-
8 and ZIF
-
8@DOX
-
TPP NPs.
TEM images of ZIF
-
8 NPs (
A
) and ZIF
-8@DOX-
TPP NPs (
B
). Scale bars, 500 nm.
Typical motion
trajectories (over 20 s) (
C
), MSD (
D
), Deff (
E
), and speed (
F
) of ZIF
-8@DOX-
TPP NPs in PBS with various H
2
O
2
concentrations. Comparison of typical motion trajectories
(over 20 s) (
G
) and MSD (
H
) of ZIF
-8 and ZIF-
8@DOX-
TPP NPs in PBS with
100 μM H
2
O
2
. (n
=
15; mean ± SEM).
F
ig. S12. Representative fluorescence images showing mitochondrial colocalization in T24
cells upon incubation with ZIF
-
67@DOX
-
TPP
nan
orobots for 2, 4, 6, 12 hours and
corresponding PCC value
(n = 5; mean ± SD).
Scale bar, 10 μm.
Fig.
S13. Representative fluorescence images showing mitochondrial colocalization in 4T1
cells upon various incubations for 12 hours, including DOX, DOX
-
TPP, ZIF
-
8@DOX, ZIF
-
8@DOX
-
TPP, ZIF
-
67@DOX, and ZIF
-
67@DOX
-
TPP nanorobots.
(n=5, mean ± SD). Scale
bars, 10
μm
.
F
ig. S14.
JC
-
1 staining images and calculated J
-
aggregate/J
-
monomer ratios of T24 bladder
cancer cells that incubated upon various conditions for 8 hours
(n = 5; mean ± SD)
.
Scale bars,
20 μm.
Fi
g. S15. The effect of H
2
O
2
decomposition on the anticancer efficacy of DOX
-
TPP after
incubation with T24 cells for 48 hours
(n=3, mean ± SD).
F
ig. S16 . The viability of
4T1 breast cancer cells
(
A
)
and DOX
-
resistant BIU
-
87/ADR bladder
cancer cells
(
B
)
after incubation with various treatments for 48 hours
(n = 3; mean ± SD).
Fi
g. S17. I
n
vitro evaluation of ZIF
-
67@DOX
-
TPP nanorobots
for metastasis inhibition in
4T1 breast cancer cells.
(A) Optical images showing in vitro wound healing assay and (B)
corresponding wound closure percentages. The ‘wound’ (cell gap) was built by a straight scratch
across 4T1 cancer cells (0 h). The wound
closure rate was examined after incubation with
nanorobots and other control groups for 12 hours. (n = 5; mean ± SD). Scale bar, 100 μm. (C)
Images of invaded 4T1 cells across the Matrigel barrier after treatment with nanorobots and other
control groups in
the upper chamber of transwell assay and (G) corresponding invasive contents
(n = 5; mean ± SD). Scale bar, 200 μm. ***
P
< 0.001, ****
P
< 0.0001; One-
way ANOVA.
F
ig. S18 . Images of resected subcutaneous T24 tumors at the endpoint of the treatment
course with nanorobots and other constructs.
Scale bar, 2 cm.
Fig
. S19. Immunohistochemical staining of resected liver slices from the group of PBS
(
A
)
and ZIF
-
67@DOX
-
TPP nanorobot
(
B
)
and T24 tumor slice of the nanorobot group
(
C
)
after
the treatment course of subcutaneous tumor model.
HLA
-
DRA pAb
was used to label the
expressed HLA
-
DRA on the surface of
human-derived
T24 cancer cells (Brown)
. Scale bar, 100
μm.
Fig. S20 . H
＆
E staining of histological sections from main organs, including heart, liver,
spleen, lung, and kidney, resected at the endpoint of the treatment course from T24 tumor
-
bearing mice that incubated with ZIF
-
67@DOX
-
TPP nanorobots and other control groups.
Sca
le bar, 100 μm.