Synthesis of folic acid functionalized terbium-doped dendritic fibrous nano-silica and Interaction with HEK 293 normal, MDA breast cancer and HT 29 colon cancer cells

Article abstract of DOI:10.1002/jmr.2871

A novel folic acid functionalized terbium-doped dendritic fibrous nanoparticle (Tb@KCC-1-NH₂-FA) with high surface area was synthesized using a novel hydrothermal protocol. In the present work, we report the fluorescent Tb-doted nanomaterial wi

Full text of DOI:10.1002/jmr.2871

Received: 2 April 2020
DOI: 10.1002/jmr.2871
Revised: 28 May 2020
Accepted: 6 June 2020
R E S E A R C H A R T I C L E
Synthesis of folic acid functionalized terbium-doped dendritic
fibrous nano-silica and Interaction with HEK 293 normal, MDA
breast cancer and HT 29 colon cancer cells
1
1
2
Sajjad Azizi | Jafar Soleymani
| Nasrin Shadjou
1
Pharmaceutical Analysis Research Center,
Abstract
Tabriz University of Medical Sciences,
Tabriz, Iran
A novel folic acid functionalized terbium-doped dendritic fibrous nanoparticle
Tb@KCC-1-NH -FA) with high surface area was synthesized using a novel hydro-
2
Department of Nanotechnology, Faculty of
(
2
Science and Chemistry, Urmia University,
Urmia, Iran
thermal protocol. In the present work, we report the fluorescent Tb-doted nano-
material with emission wavelength at 497 nm which confirms the formation of
Correspondence
Nasrin Shadjou, Department of
Nanotechnology, Faculty of Science and
Chemistry, Urmia University, Urmia, Iran.
Email: n.shadjou@urmia.ac.ir
Tb@KCC-1-NH -FA. Synthesized nanoparticles were investigated through transmis-
2
sion electron microscope, field emission scanning electron Microscopy, Fourier trans-
form infrared spectra, Brunauer-Emmett-Teller, energy dispersive X-ray, Zeta
potential and particle size distribution values and AFM (Atomic force microscopy)
techniques. Specially, our desired nanomaterial which has FA moieties on the surface
of Tb@KCC-1-NH2-FA where interact with folate receptor (FR) which there is on the
surface of the various cancer cells. For this purpose, fluorescence microscopy images
were used to prove the uptake of FA based nanomaterial with FR-positive MDA
breast cancer and HT 29 colon cancer cells. Also HEK 293 normal cells as FR-
negative cells verified the specificity of our desired nanomaterial toward the FR-
Funding information
Tabriz University of Medical Sciences;
University of Tabriz
2
positive cells. The cytotoxicity survey of Tb@KCC-1-NH -FA was examined by MTT
assays against MDA breast cancer, HT 29 colon cancer and HEK 293 Normal cell
lines which confirmed their biocompatible nature with any significant cytotoxic
effects even for concentration higher than 900 μg/mL which could be used as a non-
toxic catalyst or carrier in biological ambient. Hence, Tb@KCC-1-NH -FA were syn-
2
thesized using green and hydrothermal method; the process was simple with good
productivity and desired nanocomposite was non-toxic.
K E Y W O R D S
biological assay, breast cancer, cell viability, chemical binding, colon cancer, cytotoxicity,
fibrous nano-silica
3
-6
7
1
|
INTRODUCTION
synthesis, targeting of drugs and medical diagnosis and cancer cells
8
detections. Among them, Fibrous nano-silica (KCC-1) has become a
Recent advances in nanomaterials and nanotechnology have offered
new solutions to several problems in the field of biomedicine. Porous
nanomaterials with large pore volumes, high surface area-to-volume
ratio and other characteristics have been developed because of their
useful system in the medicinal researches because of its physicochemical
properties such as stable framework, low-toxicity, mesoporous structure,
easily modified surface, biocompatible and ultrasensitive detection of
analysts and controlled-release of the captured particles. All these prop-
erties cause to use KCC-1 based on folic acid (FA) nanomaterials as a
1,2
wide range of uses in the field of drug delivery, catalysis and organic
J Mol Recognit. 2020;e2871.
wileyonlinelibrary.com/journal/jmr
© 2020 John Wiley & Sons Ltd
1 of 9
https://doi.org/10.1002/jmr.2871

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AZIZI ET AL.
particularly potential material for medicinal requests such as imaging,
99.0%), triethanolamine (TEA, 99%), (3-aminopropyl) triethoxysilane
(APTES, 98%), tetraethyl orthosilicate (TEOS, 99%), 1-ethyl-3-(3-dimethyl-
aminopropyl) carbodiimide (EDC, 99%), 3-(4,5-dimethylthiazol-2-yl)-
2,5-diphenyltetrazolium bromide (MTT) and phosphate buffer solution
(PBS) were obtained from Merck Company. (Germany). Hydrochloride
acid (HCl), ethanol and sodium hydroxide (NaOH) were obtained from
Scharlau (Barcelona, Spain). Toluene, dimethylsulfoxide (DMSO), hexanol,
cetyltrimethylammonium bromide (CTAB) and rhodamine B (Rh B) were
purchased from Merck (Germany). Also, Roswell Park Memorial Institute
1640 growth medium (RPMI) was obtained from Gibco BRL Life Technolo-
gies (New York). Trypsin-EDTA (25%), fetal bovine serum (FBS), and
streptomycin-penicillin were purchased from Biowest (Nuaillé, France).
HEK 293, HT 29 and MDA cell lines were purchased from National Cell
Bank of Iran (NCBI).
8-10
sensing and therapeutics.
Regarding these facts, the different shapes
of the silica-based materials (porous, dendritic, wire like etc.) and their
biocompatibility offer strong cancer cells detector. Also, supplementary
modification of KCC-1 based nanocomposites can develop the use of
11,12
these nanoparticles for bio-analytical uses.
KCC-1) based dendritic nanocomposites are stable even at more intense
situation of pressures and excessive temperatures which their porous
In addition, Nano silica
(
13,14
morphology remained unaltered.
Moreover, production and purifi-
cation of nano-silica (KCC-1) based dendritic nanomaterials are easy
which could be kept in room temperature for long time without any alter-
15
ations in their properties. In original, mesoporous silica materials have
no well-known biological activities but they can recognize by the immu-
nology system as foreign compounds and collected in the liver and
16
spleen. Huang et al evaluated the cytotoxic activity of KCC-1 based
nanomaterials on A2780 human ovarian cancer cell lines and they noted
no important cell death at concentrations 107.5 ± 6.4% μg/mL during
2.2
|
Instruments
17
4
8 hours. There are many approaches for the detection of various can-
1
8
19
cer cells which were reported by Huang et al, Mahmoud et al, Chinen
Transmission electron microscopy (TEM) images were recorded by a
Carl Zeiss LEO 906 electron microscope operated at 100 kV
(Oberkochen, Germany). Energy dispersive X-ray (EDX) and Field
Emission Scanning Electron Microscopy (FESEM) were conducted on
FEG-SEM MIRA3 TESCAN (Brno, Czech Republic). Fourier transform
infrared (FTIR) spectra were obtained with a Bruker Tensor 27 spec-
trometer (Tokyo, Japan). Brunauer-Emmett-Teller (BET) was recorded
by Micromeritics NOVA 2000 apparatus at 77 K using nitrogen as the
adsorption gas (Florida). Particle size distribution values and zeta poten-
tial were measured by Malvern particle size analyzer (Malvern, UK).
MTT survey results were investigated with a microplate reader Aware-
ness Technology (Florida, USA) at 570 nm. Fluorescence spectra were
obtained by a Jasco FP 750 spectrofluorometer (Tokyo, Japan). Fluores-
cence images were recorded on a Model CytationTM 5 (Winooski) and
Bh2-RFCA fluorescence Olympus microscope (Tokyo, Japan).
2
0
21
22
et al, Islam et al, Smith et al, and so on. However, several methods
have disadvantages such as instabilities, low specificity, with difficult
product approaches which almost limited their applications in biological
uses. It is undeniable that many researchers verified the interaction of FA
based materials with FR-positive cancer cells are useful in cancer cells
8
,23,24
detection which detected by imaging, sensing and therap.
Terbium (Tb) is a rare earth element which extensively used in
biological ambient and has been evaluated on toxic effects in several
25,26
studies.
Some researchers have publicized that Tb nanoparticles
27
have poor toxicity and low antitumor activity. Also, this fluorescent
lanthanide element (Tb) have been used for investigation of biological
systems, DNA and RNA hybridization assays, immunoassays, chro-
28-30
matographic detections and quantification of organic compounds.
Based on these researches, we now report the first production of a
florescence based folic acid functionalized terbium-doped dendritic
2
fibrous nano-silicas (Tb@KCC-1-NH -FA) as a non-toxic material using a
hydrothermal method and survey its cytotoxicity against MDA breast
cancer, HT 29 colon cancer and HEK 293 normal cell lines and first intro-
2.3
|
preparation of nanocomposites
| Synthesis of Tb@KCC-1-NH2
duced it to the field of biomedicine. Moreover, Tb@KCC-1-NH
2
-FA pos-
2.3.1
sessed cancer cell-targeting abilities which FA in its structure could
interact with folate receptors in cancer cells that recorded by fluores-
cence microscopy images also its anticancer activity is poor. In addition,
The preparation procedure of Tb doped in the surface of KCC-1
nanoparticles is explained as follows: Briefly, 5 mL of NaOH (2 M) and
1.0 gr of CTAB and 230 mL of deionized water were mixed and soni-
3+
we coated the Tb ions with porous nanoparticles as a florescent metal
to increase the florescence properties of nanocomposite for spectropho-
tometric studies. So, the novel Tb-doted nanoparticles are expected to
have low cytotoxic effects for biological ambient.
3 2
cated for 20 minutes. Then, 0.5 g of TbCl .6H O and 5.2 mL of TEOS
were combined drop-wise in the reaction ambient. Next, the resultant
ꢀ
solution was refluxed at 90 C for 1.5 hours. Afterward, obtained precipi-
tations were filtrated and immediately washed tree times with deionized
water and absolute ethanol. Eventually, as prepared participated was cal-
ꢀ
2
2
|
EXPERIMENTAL SECTION
| Materials
cinated at 550 C for 5 hours to eliminate the extra CTAB in the oven.
In order to synthesis of Tb@KCC-1 nanoparticle functionalized
with amine groups, firstly, 260 mg of Tb@KCC-1 nanocomposite was
added to the 10 mL of HCl solution. Then, 6 mL of ethanol and 790 μL
of APTES were added to the solution and stirred at room temperature
for 6 hours. Next, the obtained suspension was centrifuged and washed
.1
Terbium chloride (6.H
2
O) and FA were purchased from Merck (Germany)
and Mehr Darou (Tehran, Iran), respectively. N-hydroxysuccinimide (NHS,

AZIZI ET AL.
3 of 9
several times with absolute ethanol. Then, washed Tb@KCC-1-NH
2
and 5% CO
2
) for about 4 days to reach desired confluence of cells.
white solids were dried at room temperature.
Then, the supernatant of flasks was discarded and washed with buffer
solution (pH = 7.4). After addition of trypsin-EDTA, the flasks were
incubated for about 10 minutes to detach the HEK 293 normal and
MDA breast cancer cells from the flask surface. Next, cell suspensions
were centrifuged at 1500 rpm for about 10 minutes to remove
unreacted trypsin-EDTA. Eventually, the cells were introduced in
2
.3.2
|
Synthesis of Tb@KCC-1-NH -FA
2
For the synthesize of FA-NHS by EDC/NHS, firstly, 70 mg of folic
acid, was mixed to with 15 mL of DMSO and 160 μL of TEA and
stirred vigorously. Then, 13 mg of NHS and 11 mg of EDC were
2
refresh RPMI media to count with a hemocytometer (0.0025 mm ).
2
Toxicity activities of Tb@KCC-1-NH -FA particles towards cancer
ꢀ
added and stirred vigorously at 37 C for 20 hours. Next, TEA was
and normal cells were investigated by MTT survey protocol. To do this
4
eliminated by vaporization and synthesized folic acid-NHS was kept in
a refrigerator for next stage of synthesis.
assay, about 1.0 × 10 of cancer and normal cells were seeded into
each well of a 96-well plate and incubated for about 24 hours. After-
ward, cancer and normal cells were treated with various concentra-
2
In continue, 180 mg of Tb@KCC-1-NH nanoparticles were mixed
with 30 mL of PBS (pH 7.0, 55 mM) and next 15 mL of the synthesized
FA-NHS solution was mixed with the solution. Next, the mixture of
2
tions of Tb@KCC-1-NH -FA (0-900 μg/mL, separately) and incubated
for a desired of 24 hours. Next, MTT solution (3.0 mg/mL) was added
to the wells and incubated for another 5 hours. Then, the media were
replaced by DMSO (about 200 μL) and then incubated for 45 minutes.
Finally, a plate reader was utilized to record the absorbance of the
wells (570 nm). In the MTT survey, nanomaterial-untreated cancer
cells were considered as a negative control.
Tb@KCC-1-NH
2
and prepared folic acid-NHS was stirred at room tem-
-FA white-milky pre-
perature for 6 hours. After that, Tb@KCC-1-NH
2
cipitate was filtered and washed several times with deionized water
ꢀ
and dried at 60 C. The schematic procedures of Tb@KCC-1-NH
2
-FA
nanoparticles preparation was demonstrated in Scheme 1.
2
.4
|
Cell culture and MTT assay
2.5
|
Cell uptake study
HEK 293 normal and MDA breast cancer cells were seeded into
2
To verified the suitable uptake of Tb@KCC-1-NH -FA nanoparticles
2
5-mL flasks with RPMI-1640 medium with 10% FBS, and 1% penicil-
with FR-positive cancer cells, fluorescence imaging was carried out as
a main technic for qualitative uptake distinguish. Firstly, MDA breast
ꢀ
lin/streptomycin and then incubated in a misted atmosphere (37 C
2
SCHEME 1 The synthesis procedure of Tb@KCC-1-NH -FA

4
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AZIZI ET AL.
FIGURE 1 TEM images of KCC-1-NH
F-H)
2
-FA (A, B), Tb@KCC-1-NH
2
2 2
-FA (C, D), FE-SEM images of KCC-1-NH -FA (E) and Tb@KCC-1-NH -FA
(
5
z + 3
cancer and HT 29 colon cancer cells (5 × 10 ) were seeded to the
cell-culture plate and incubated for 24 hours. Then, several concentra-
tion of Tb@KCC-1-NH2-FA nanoparticles were added to the all
1) ]
n
nanomaterials. Which n and z mean to the amount of KCC-1
and charge of KCC-1 composite, respectively.
2
EDX analyze shows the atomic composition of the KCC-1-NH -
6
-well of plate and incubated for 2, 5, 10, 15 and 24 hours. Next,
2
FA and Tb@KCC-1-NH -FA (Figure 2) which combinations percent
RPMI throw away and PBS solution were used for washing the wells.
Afterward, the uptakes of Tb@KCC-1-NH2-FA nanomaterials with
MDA and HT 29 cancer cells were studied with the fluorescence
Olympus microscope to record fluorescence images.
are seen in Table 1. After functionalization of the composite with ter-
bium ions its mass percent was excelled which confirm the exact join
of terbium ions into the three-dimensional composition of KCC-
1-NH
ions is near 1% of the prepared Tb@KCC-1-NH
For the recognition of the surface of Tb@KCC-1-NH
2
composite. Moreover, it revealed that the percent of terbium
2
-FA.
2
-FA nano-
3
3
|
RESULTS AND DISCUSSION
material we used the AFM images which shown in Figure 3. As could
be seen, the images conformation confirmed the FESEM and TEM
images information which size and the fibrous construction of the raw
KCC-1 were altered by doping of Terbium ions.
.1
|
Characterization of Tb@KCC-1-NH -FA
2
nanoparticles
FTIR analyze was employed to confirm the correctness synthesis
TEM images were used to realize the particle size and shape of KCC-
2
of Tb@KCC-1-NH -FA. As seen in Figure 4, the typical transmittance
1
-NH
Figure 1A-D). Therefore, the TEM images revealed an important dif-
ference in the formation of Tb@KCC-1-NH -FA nanomaterial from
2
-FA composite and Tb@KCC-1-NH
2
-FA nanomaterials
peaks of silica-based composites could be seen in the range of 1020
to 1110 cm− signifying the asymmetric stretching of Si-O-Si mode.
Also, Si-OH spectrum which located at 960 cm− denotes the asym-
metric bending and stretching vibration. The spectra could be seen at
1
(
1
2
fibrous to the nanocomposite shaped with changed crystal morphol-
ogy. Although terbium ions doted to the nanocomposite, the porosity
the range 452 and 411 cm− are related to Tb-O bonds. However,
1
32
1
of the Tb@KCC-1-NH
FESEM images shows morphological and structural features of
the KCC-1-NH -FA and Tb@KCC-1-NH -FA with different scales
Figure 1E-H). As seen, the fibrous-like structure of KCC-1-NH -FA is
altered to the crystal-like in Tb@KCC-1-NH -FA with different sizes
2
-FA composite does not change.
peak at around 1500 cm− is related with amide II bonds between car-
boxyl's of FA molecules and amine's of KCC-1-NH
The BJH, BET analyzes and the N adsorption/desorption iso-
therms of Tb@KCC-1-NH -FA and KCC-1-NH -FA were applied to
2
.
2
2
2
(
2
2
2
2
evaluate the porous essence, pore volume and specific surface area
(Figure S1). As shown in Table 2, the surface area, pore volume and
and shapes. Actually, the addition of terbium ions effect to the coagu-
lation of the mono-dispersed KCC-1 molecules to the composite with
enormous amount of KCC-1 components to synthesize of [Tb(KCC-
pore size of Tb@KCC-1-NH
2
-FA were reduced with adding terbium.
Also, the porous volumes of KCC-1-NH
2
-FA were changed from 1.3 to

AZIZI ET AL.
5 of 9
FIGURE 2 EDX analysis of
2
Tb@KCC-1-NH -FA (A) and KCC-
1
-NH -FA (B)
2
TABLE 1 EDX analysis data for KCC-NH
2
-FA and Tb@KCC-NH
2
-
3
0
.25 cm /g for Tb@KCC-1-NH
2
-FA nanomaterial which proved that
FA with weight and atomic percentages
3+
the surface of composite was excellently occupied with Tb ions. Also,
a
KCC-NH2-FA
Tb@KCC-NH2-FA
the data confirmed by pore size values. As could be seen, the revealed
3+
results proved that surface area in the nanocomposite with Tb ions
Element
W%
2.46
A%
3.88
W%
A%
2
doping has not change and it reduced from 397 to 311 m /g which is
C
15.57
2.90
23.54
3.77
2
as an improvement of the prepared Tb@KCC-1-NH -FA to prepare
N
1.43
57.46
38.65
0.00
1.94
68.09
26.09
0.00
14,33
great range of doping terbium and surface area.
The zeta potential analysis of Tb@KCC-1-NH
was determined and contrasted with KCC-1-NH
O
41.89
38.69
0.95
47.56
25.02
0.11
2
-FA nanocomposite
Si
2
-FA nanomaterial at
Tb
Total
pH 7.5 to checked the influence of Tb ions which doped on the surface
charge and show the exact doping of KCC-1-NH -FA nanocomposite
with Tb ions. Thus, both of them (Tb@KCC-1-NH -FA and KCC-
1-NH -FA) exhibited positive external charge in contrasted with pure
100.00
100.00
100.00
100.00
2
3+
a
31
2
Synthesis procedure was reported at our previously reported paper
which is different from the synthesis approach of Tb@KCC-NH2-FA.
2

6
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AZIZI ET AL.
2
FIGURE 3 AFM topography images of Tb@KCC-1-NH -FA
2
FIGURE 4 FTIR spectra of the KCC-1, Tb@KCC-1-NH -FA
2
FIGURE 5 Fluorescence spectra of Tb@KCC-1-NH -FA,
and FA
Tb@KCC-1-NH2, Tb and FA
TABLE 2 Surface area, average pore size, and pore volume of
Tb@KCC-1-NH
-NH which generally concerned to cavity and surface functionalization
of the nanomaterial with FA molecules and Tb ions.
2
-FA increased in compared with the KCC-1 and KCC-
KCC-1, KCC-1-NH
2
and KCC-1-NH
2
-FA
1
2
3+
34
Pore size
Pore volume
Surface Area
a
3
b
2
Material type
(nm)
(cm /g)
(m /g)
In addition, fluorescence analyze of the prepared composites was
investigated to recognize the resource fluoresce peaks and the efficacious
KCC-1-NH
2
-FA
13.2
1.3
397
311
3+
Tb@KCC-1-NH
2
-FA
3.9
0.25
preparation of nanomaterial. As could be seen in Figure 5, Tb ions and
Tb@KCC-1-NH nanomaterial have some original emission peak at
491, 545, 585 and 622 nm where are referred to D
a
Pore size was calculated by BET.
2
b
5
7
5
7
Pore volume determined by BJH.
4
! F
6
, D
4
! F5,
5
7
5
7
35,36
D
4
! F
4
and D
4
! F
3
electron transmission, sequential.
However,
3+
the spectral peaks of Tb@KCC-1-NH
2
were altered to the Tb ion fluores-
3+
3
+ 33
KCC-1 nanoparticles, which were mainly as the result of FA and Tb
.
cence peak after functionalization with Tb ions. The related spectral peak
Also, the positive surface charge of synthesized Tb@KCC-1-NH
2
-FA
of Tb@KCC-1-NH
trum (450 nm) to 497 nm where showed the successful and accurate func-
tionalization of Tb@KCC-1-NH nanocomposite with FA particles. On the
2
-FA nanocomposite was altered from FA specific spec-
3+
nanocomposite confirm the fixing FA and Tb ion groups on the superfi-
33
cial spaces of the fibrous composites, respectively. Moreover, DLS
analysis of Tb@KCC-1-NH -FA confirmed that its hydrodynamic dimen-
sion was 820 nm which has agreement with AFM pictures. The size of
2
3+
2
other hand, the Tb ions could absorb with hydroxyl groups of KCC-1
porous materials to expand their spectroscopic properties.

AZIZI ET AL.
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2
FIGURE 6 MTT cell viability assay of synthesized Tb@KCC-1-NH -FA on HT-29 cancer cell line were measured at 24 hours
3
.2
|
Evaluation of cytotoxicity activity of
4
|
CONCLUSION
Tb@KCC-1-NH -FA
2
2
In the present research, Tb@KCC-1-NH -FA nanocomposite was
MTT survey is a well-known colorimetric test where using the reduc-
tion of 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
by the succinate dehydrogenase enzyme of alive cell mitochondria.
MTT assay is utilized for cytotoxic investigations (minimum of living
cell) of materials. The mechanism action of MTT assay are including
the MTT color entrance in to the normal and cancer cells and pierces
into the mitochondria, which reduced to the insoluble purple color
formazan. For this purpose, we dissolve the mention formazans in
DMSO, and its absorbance is recorded at 500 to 600 nm wavelength
through a spectrophotometer. In original, these kind of color materials
successfully synthesized using hydrothermal method. The fluorescence
signal of this Tb-doted nanomaterial was emitted at 497 nm wavelength.
2
Remarkable feature of Tb@KCC-1-NH -FA nanocomposite including
high mesoporous surface area, easily modified surface, excellent biocom-
patible properties and desirable physical and chemical stability. The
morphology and structure of synthesized Tb@KCC-1-NH
investigated which the results approve the formation of Tb@KCC-
1-NH -FA porous nanomaterial with an average molecule size (3.9 nm).
2
-FA were
2
The toxicity assay of nanocomposites on the normal, colon and breast
cancer cell line did not have significant toxicity even for 900 μg/mL con-
(
MTT) which are a tetrazolium compounds reductions are related on
2
centration. Hence, it suggested that synthesized Tb@KCC-1-NH -FA
metabolism of checked cells where if this metabolism be in low rate,
can be utilized in biological mediums. Also, Tb@KCC-1-NH2-FA
nanoparticles were used as a proper desired uptake to FR-positive
cancer cells to specific determination of MDA breast and HT 29 colon
cancer cells.
37,38
the reduction of MTT will be low too and vice versa.
In our work, MTT measurement was used to studding the cell via-
bility of Tb@KCC-1-NH -FA treated HEK 293 normal, HT 29 colon
and MDA breast cancer cells. Figure 6 include the viability percent of
numerous concentration (0-900 μg/mL) of Tb@KCC-1-NH -FA. It is
clear that there is no remarkable toxic result for synthesized
2
2
ACKNOWLEDGEMENTS
This Research was supported by University of Tabriz and Tabriz
University of Medical Sciences. All authors have read, approved
and made substantial contributions for the manuscript. None of
the original material contained in this manuscript has been previ-
ously published nor is currently under review for publication
elsewhere.
2
Tb@KCC-1-NH -FA even for high concentration (900 μg/mL) after
2
4 hours incubation and cell viabilities are more than 82%. Thus,
2
Tb@KCC-1-NH -FA nanomaterials have low cytotoxic result on nor-
mal and cancer cells which could be utilized as a biocompatible nano-
material in biological mediums.

8
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AZIZI ET AL.
CONFLICT OF INTEREST
16. Liu T, Li L, Fu C, Liu H, Chen D, Tang F. Pathological mechanisms of
liver injury caused by continuous intraperitoneal injection of silica
nanoparticles. Biomaterials. 2012;33:2399-2407.
The authors declare no conflict of interests and/or commercial prod-
ucts or companies.
1
7. Huang X, Tao Z, Praskavich JC Jr, et al. Dendritic silica nanomaterials
KCC-1) with fibrous pore structure possess high DNA adsorption capacity
(
and effectively deliver genes in vitro. Langmuir. 2014;30:10886-10898.
8. Huang Y, Liu Q, Wang Y, et al. Gold nanorods functionalized by a gluta-
thione response near-infrared fluorescent probe as a promising nan-
oplatform for fluorescence imaging guided precision therapy. Nanoscale.
ORCID
1
Jafar Soleymani
Nasrin Shadjou
https://orcid.org/0000-0002-8316-5104
https://orcid.org/0000-0001-5961-3686
2
019;11:12220-12229.
1
9. Mahmoud ME, Osman MM, Yakout AA, Abdelfattah AM. Green
nanosilica@folic acid (VB9) nanocomposite for engineered adsorptive
water remediation of bivalent lead, cadmium and copper. Powder Tech.
2019;344:719-729.
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