RSC Advances

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Cell penetrating peptide decorated magnetic

Cite this: RSC Adv., 2022, 12, 11708
porous silicon nanorods for glioblastoma therapy
and imaging†
Arnaud Chaix, ‡a Audrey Griveau,‡b Thomas Defforge,a Virginie Grimal,a Brice Le
Borgne,a Gaël Gautier*a and Joël Eyer*b

Glioblastoma multiforme (GBM) is the most malignant primary brain tumor of the central nervous system.
Despite advances in therapy, it remains largely untreatable, in part due to the low permeability of
chemotherapeutic drugs across the blood–brain barrier (BBB) which significantly compromises their
effectiveness. To circumvent the lack of drug efficiency, we designed multifunctional nanoparticles
based on porous silicon. Herein, we propose an innovative synthesis technique for porous silicon
nanorods (pSiNRs) with three-dimensional (3D) shape-controlled nanostructure. In order to achieve an
efficient administration and improved treatment against GBM cells, a porous silicon nanoplatform is
designed with magnetic guidance, fluorescence tracking and a cell-penetrating peptide (CPP). A
NeuroFilament Light (NFL) subunit derived 24 amino acid tubulin binding site peptide called NFL-
TBS.40-63 peptide or NFL-peptide was reported to preferentially target human GBM cells compared to
healthy cells. Motivated by this approach, we investigated the use of magnetic-pSiNRs covered with
superparamagnetic iron oxide nanoparticles (SPIONs) for magnetic guidance, then decorated with the
NFL-peptide to facilitate targeting and enhance internalization into human GBM cells. Unexpectedly,
under confocal microscope imaging, the internalized multifunctional nanoparticles in GBM cells induce
a remarkable exaltation of green fluorescence instead of the red native fluorescence from the dye due
Received 24th January 2022
Accepted 30th March 2022
to a possible Förster resonance energy transfer (FRET). In addition, we showed that the uptake of NFL-
peptide decorated magnetic-pSiNRs was preferential towards human GBM cells. This study presents the
DOI: 10.1039/d2ra00508e
fabrication of magnetic-pSiNRs decorated with the NFL-peptide, which act as a remarkable candidate to
rsc.li/rsc-advances treat brain tumors. This is supported by in vitro results and confocal imaging.

their low specicity to the tumor, possibly causing irreversible

Introduction side effects and the risk of developing resistance to the treat-
Glioblastoma multiforme (GBM) is extremely invasive and ment.4 Moreover, the delivery of drugs into the brain can be
considered among the deadliest brain cancers.1 GBM is a high- limited by the low permeability of the blood–brain barrier
grade (grade IV) glioma according to the World Health Orga- (BBB), which causes the inefficiency of conventional therapeu-
nization, and is fast growing, spreads within the brain and may tics such as chemotherapeutic drugs. There is a dire need to
come back even if extensively treated. In 2018, the National search for new GBM treatment strategies and nowadays nano-
Institute of Health (NIH) predicted around 24 000 new cases of medicine appears as one of the most promising approaches.
GBM brain cancer in the United States with a constant increase. During the past few decades, nanomedicine has emerged as
Nowadays, the standard treatment required for GBM is surgical an alternative strategy to overcome the limitation mentioned
resection followed by radiotherapy and chemotherapy.2 above and address effective anti-cancer treatments.5 However,
However, the recovery of patients with GBM is less than 5% over to improve the GBM treatment, the development of new thera-
5 years.3 The persistent limitations for such treatments include peutic platforms such as functionalized nanoparticles (NPs) has
become crucial to cross the BBB, reach the brain tumor and
release an optimal dose of drugs. Interestingly, the development
a
GREMAN UMR-CNRS 7347, INSA Centre Val de Loire, Université de Tours, Tours, of those targeting NPs for drug delivery have certain advantages
France. E-mail: gael.gautier@univ-tours.fr such as the prevention of premature release or drug degrada-
b
MINT, INSERM, CNRS, SFR-ICAT, UNIV Angers, 49000 Angers, France. E-mail: joel.
tion or an increased cellular uptake. In fact, a wide range of
eyer@univ-angers.fr
nanoparticles have been developed and studied as anticancer
† Electronic supplementary information (ESI) available. See
https://doi.org/10.1039/d2ra00508e therapeutics against GBM cells including lipidic nano-
‡ These authors contributed equally. capsules,6,7 polymeric NPs,8,9 mesoporous silica nanoparticles

11708 | RSC Adv., 2022, 12, 11708–11714 © 2022 The Author(s). Published by the Royal Society of Chemistry
Paper RSC Advances

(MSN),10,11 and Metal–Organic Frameworks (MOFs) NPs.12,13 cells instead of healthy human cells.7 To this mean, the physico-
Among these materials, porous silicon nanoparticles (pSiNPs) chemical properties of the functionalized nanoparticles were
are being considered as especially promising owing to their fully characterized and in vitro studies, cellular internalization
physico-chemical properties, biocompatibility, biodegradability of the nanoparticles by transmission electron microscopy (TEM)
and the low toxicity of the degradation by-products in vivo.14 In and confocal microscope imaging were performed.
particular, orthosilicate, which is non-toxic and inert for human
cells, is the major degradation by-product of pSi.15 Besides, Results and discussion
silicon is an oligo-element naturally present in signicant
amount in the human tissues, whether bone, organic or Preparation of porous silicon nanorods (pSiNRs)
connective. One of the main advantages of pSi materials pSiNPs size, shape, pore size and surface chemistry are of
compared to other nanomaterials is the possibility of accurately highest importance to the success of the study. Herein, we
adjusting the structures, shapes, porosity, and pore size summarize the elaboration of pSiNRs controlled in shape and
according to the intended application. For instance, pSi mate- size using a two steps process. To this aim, pSiNRs were fabri-
rials can exhibit high specic surface area (up to 1125 m2 g
1), cated from an innovative way combining electrochemical
high porosity (in the 30–95% range) and large pores size.16–18 etching14 of silicon wafer followed by Metal-assisted Chemical
These morphological properties combined with the silicon Etching (MaCE)45,46 obtaining a nanorod shape depicted in
surface chemistry enable versatile surface modication oppor- Fig. 1. This fabrication technique takes its origin to the need of
tunities and therefore a large range of targeting ligand func- the synthesis of particles with homogeneous dimensions in 3D
tionalizations such as peptides,7,19 antibodies,20,21 sugars,22,23 over the single direction pSiNPs that we actually nd in most of
and molecules.24 Moreover, pSiNPs have been extensively the scientic literature.47,48 First, pSi was fabricated by electro-
investigated for drug encapsulation with an exceptional loading chemical etching of boron doped p type silicon wafer in an
capacity25 and a variety of therapeutic drugs for instance electrolytic solution of deionized water, acetic acid, and
chemotherapeutics,26,27 oligonucleotides,19 and nucleic acid.28,29 hydrouoric acid (5% HF). During the electrochemical etching
To our knowledge some studies have already been reported leading to pSi formation, periodic “perforations” were carried
using pSiNPs as a nanoplatform for targeting and therapy of out alternating “low” and “high” current density periods (for
GBM.30–33 Subsequently, composite nanomaterials based on detailed information: see the ESI†). Hence, porous layers with
pSiNPs have been also reported for biological applications in alternating low and high porosity strata in depth were obtained
particular with the decoration with smaller size nanoparticles with a periodicity of 200 nm. Scanning electron microscopy
typically gold nanoparticles NPs,34 silver NPs,35 and super- (SEM) conrmed the observation of this stratied layers
paramagnetic iron oxide NPs (SPIONs).27 To this aim, we (Fig. S1, ESI†). Thereaer perforated pSi layer was subjected to
focused our effort on novelties including (3D) shape-controlled MaCE treatment in presence of HF and Ag+ ions in order to
nanostructure fabrication of pSi and multiple functionalities of fabricate perforated porous silicon nanowires (pSiNWs) (Fig. S2,
the nanoplatform such as magnetic guidance, uorescent ESI†). The Fig. S3 in the ESI† revealed the different steps of the
tracking, and cell-penetrating peptide (CPP) carrying, the latter preparation of the perforated pSiNWs on 600 silicon wafers. The
leading to GBM cellular death. Then, SPIONs were used to pSiNWs were then mechanically peeled off the parent substrate
decorate the fabricated nanorods and appear as a useful and dispersed into deionized (DI) water solution (Fig. S4, ESI†)
strategy to reach the human GBM cells by transportation under and fractured by ultrasonication for 24 hours (Fig. S5, ESI†).
magnetic eld. Specically, SPIONs already demonstrated to be During the ultrasonication step, the nanowires preferentially
a powerful nanomaterial for biological applications due to their break at the level of the most fragile zone (the most porous)
low-toxicity,36–38 Magnetic Resonance Imaging (MRI) contrast region of the strata therefore producing nanoparticles with
agent39,40 efficiency and their magnetic eld guidance ability.27 repeatable dimensions. From this fabrication process, the
In addition, uorescent dye (DiD, see ESI†) was loaded into the nanoparticles were calibrated in the three directions: x direc-
mesoporous structure of pSiNRs for uorescent tracking under tion (in the depth of the silicon wafer direction) during the
confocal microscope imaging. In this study, a peptide corre- electrochemical etching (stratied layer), and y and z directions
sponding to the sequence of the NeuroFilament Light subunit during the MaCE leading to nanowires formation as described
which binds tubulin (NFL-TBS.40-63 peptide for NeuroFilament on the Fig. 1. Aer sonication and a series of centrifugation,
Low Subunit-Tubulin Binding Site 40-63), also called NFL- a desired size of pSiNRs (between 250–500 nm) was obtained
peptide was used. This peptide interacts or penetrates speci- and characterized by TEM, SEM and dynamic light scattering
cally in all glioblastoma cell lines tested (rat, mouse, human (DLS). The SEM and TEM also validated the 3D controlled
and dog), and by blocking the polymerization of microtubules, nanostructures, their monodispersity and mesoporosity with an
it inhibits cell division in vitro and in vivo.41,42 To date, some average pore size of 10 to 15 nm (Fig. 2a, b, S6 and S7, ESI†). The
studies report the used of NFL-peptide as targeting agent and apparent nitrogen adsorption and desorption measurements
Eyer and co-workers pioneering work led to the development of revealed the mesoporous structure of pSiNRs with a Brunauer–
targeted-nanoparticles based on NFL-peptide.38,43,44 Indeed, Emmett–Teller surface area (SBET) of 293 m2 g
1 and a BJH
a recent study reported by Karim et al. demonstrates the use of (Barrett–Joyner–Halenda) desorption average pore diameter of
lipid nanocapsules decorated with NFL-peptide which were 10 nm (Fig. S8a, ESI†). The pSiNRs display large pore volume
preferentially internalized into human, rat and mouse's GBM and high specic surface area, suitable for graing and loading

© 2022 The Author(s). Published by the Royal Society of Chemistry RSC Adv., 2022, 12, 11708–11714 | 11709
RSC Advances Paper

Fig. 1 Schematic representation of the preparation of porous silicon nanorods (pSiNRs): (i) etching, (ii) MaCE, (iii) peel off, and (iv) ultrasonication.

Preparation of the multifunctional nanoparticles

In this research project, pSi-based nanoplatforms against
human GBM cells were designed. Eight different formulations
were prepared to determine the inuence of each new func-
tionalization on GBM cell eradication. Those eight formations
are designed as: fresh pSiNRs, the ones containing the uo-
rescent dye loaded into the mesoporous structure (pSiNRs-DiD),
the ones containing SPIONs (pSiNRs@SPIONs, pSiNRs-
DiD@SPIONs) and four with the BIOT-NFL-peptide [bio-
tinylated] or with FAM-NFL-peptide [tag with uorescein]
(pSiNRs@SPIONs-BIOT-NFL, pSiNRs@SPIONs-FAM-NFL,

Fig. 2 (a) Transmission electron microscopy image of pSiNRs. Scale

bar: 500 nm (b) scanning electron microscopy image of pSiNRs. Scale
bar: 200 nm (c) z-potential measurements at each step of modifica-
tions. Bars represent mean  SD (n ¼ 3). (d) Photography of magnetic-
pSiNRs under magnetic field.

of molecules. The (DLS) conrmed an average hydrodynamic

diameter of pSiNRs at 250 nm with a polydispersity index of
0.17, which is in agreement with the observation of TEM and
SEM images (Fig. S8b, ESI†). As expected, surface charge of the
fresh pSiNRs was determined by zeta potential (ZP) measure-
ment with negative value of
40 mV in deionized water (Fig. 2c).
The negative charge value of the pSiNRs was ascribed to the
surface oxidation during the MaCE. Additionally, powder X-ray
diffraction (PXRD) conrmed the crystalline structure of the
pSiNRs network (Fig. S8d, ESI†).
Fig. 3 Schematic representation of the nanoparticle formulations.

11710 | RSC Adv., 2022, 12, 11708–11714 © 2022 The Author(s). Published by the Royal Society of Chemistry
Paper RSC Advances

pSiNRs-DiD@SPIONs-BIOT-NFL, and pSiNRs-DiD@SPIONs- of the nanorods (Fig. S20 and S22, ESI†). The ATR-FTIR pre-
FAM-NFL) (Fig. 3). sented new intense band appeared at 2875–2920 cm
1, which
The rst step consists in graing of (3-aminopropyl)trie- corresponds to the stretching vibration of the C–H band
thoxysilane (APTES) onto fresh pSiNRs in ethanoic solution (signature of the aliphatic chain of the DiD) (Fig. S21, ESI†). The
under reux condition (Scheme S1, ESI†). The post chemical UV-vis spectroscopy presented an absorption peak at 660 nm
modication of fresh pSiNRs with an amine as a terminal group which corresponds to the absorbance pic of the DiD molecules
has been an obvious approach to provide a higher interaction (Fig. S22, ESI†). The presence of DiD dye into the framework of
with the negative charges of the SPIONs. Zeta potential and DLS the nanoparticles allows us an optical tracking into the human
measurements conrmed the attachment of the amine groups GBM cells with the help of a confocal imaging microscope.
onto the surface via silanisation reaction (Fig. 2c and S9, ESI†). Finally, CPP-magnetic-pSiNRs with or without uorescent dye
The DLS exhibits an increase of the hydrodynamic diameter were prepared as described on the Schemes S5 and S6 (ESI†),
from 250 nm to 381 nm (Fig. S9a, ESI†) attributed to a strong along for enabling selective targeting and lead cellular death.
aggregation of the pSiNRs-NH2 owed to the amine protonation The nanoparticles formulation was decorated with NFL-
in deionized water, which is in accordance with the literature.29 peptides (coupled with BIOT and FAM). Regarding the incor-
Aer the chemical graing of the APTES, the zeta potential poration of CPP to the pSiNRs, we have privileged the decora-
value change from negative charge (
40.5 mV) to positive tion onto magnetic-pSiNRs instead of covalent bonding due to
charge (+12 mV) (Fig. S9b, ESI†). ATR-FTIR spectroscopy was the loss of efficiency, which was previously demonstrated.7 The
also performed with pSiNRs-NH2 with the observation of the presence of the NFL-peptide onto the surface of the magnetic-
narrow band centered at 800 cm
1 ascribed to the amine pSiNRs was also investigated by a complete characterization
function from the APTES moiety and an increase of surface of the nanoparticles. The nanoparticles formulations contain-
oxidation compared to the fresh pSiNRs (Fig. S10, ESI†). The ing the BIOT – or FAM-NFL-peptides onto the surface were also
quantication of the attached APTES moiety was determined by fully characterized by several techniques such as TEM, SEM,
thermogravimetric analysis (TGA) with a graed amount of DLS and ZP (Fig. S23 and S27, ESI†). The DLS and ZP conrmed
0.014 mmol per mg of nanoparticles (Fig. S11, ESI†). To guide the successful decoration of the peptides with an increase in
pSiNRs under magnetic eld, 10–15 nm sized SPIONs (Fig. S12, size diameter and surface charge of the formulations (Fig. S26
ESI†) were attached to the particles (Scheme S2, ESI†). The and S27, ESI†). This is further supported by ATR-FTIR charac-
magnetic-pSiNRs (pSiNRs@SPIONs) were harvested by centri- terizations with new intense band appeared at 2920–2875 cm
1,
fugation without need of magnetic washed attraction of the assignable to the stretching vibration of the C–H band (aliphatic
non-absorbed SPIONs, which showed high affinity between chain of the NFL-peptide and aliphatic chain of DiD). In addi-
aminated-pSiNRs and SPIONs (Fig. S13, ESI†). The high amount tion, new peaks at 1525 cm
1 and 1625 cm
1 were observed,
of SPIONs decorated provide efficient transportation of the which are attributed to the functional groups of the NFL-
pSiNRs@SPIONs by using neodymium magnet (Fig. 2d and peptide (amide and carboxylic acid groups) (Fig. S28 and S30,
ESI†). The presence of pSiNRs@SPIONs was also characterized ESI†). The amount of BIOT-NFL-peptide decorated onto the
via several techniques. TEM and SEM conrmed the preferen- magnetic-pSiNRs was determined by TGA analysis with
tial self-assembly of the SPIONs onto the surface of pSiNRs-NH2 a loading amount of 0.04 mmol of BIOT-NFL per mg of nano-
and pointed out a homogeneous distribution (Fig. S14 and S15, particles (Fig. S31, ESI†). In addition, a characteristic absor-
ESI†). EDX analysis and mapping showed the presence of bance peak of uorescein from FAM-NFL-peptide was observed
SPIONs onto the surface of the nanorods with the presence of at 490 nm by UV-vis spectroscopy (Fig. S32, ESI†). Therefore,
the rays (Fe La, Fe Ka and Fe Kb) and with approximatively 15% these results suggest that the NFL-peptides were strongly
of iron element (Fig. S16 and S17, ESI†). DLS and ZP measure- absorbed to the surface of the nanoparticles. To conclude, the
ments gave additional indications on the dimensions and nal nanoconstructs maintained their magnetic properties
surface chemistry of pSiNRs@SPIONs (Fig. S18, ESI†). The aer the peptides modications as showed on the Fig. S33 in
hydrodynamic diameter of the formulations showed non- the ESI.†
aggregated nanoparticles on contrary to pSiNRs-NH2. This
phenomenon can be explained by the negative charge value of
the SPIONs attached the pSiNRs, which favors a higher solu- In vitro studies of the nanoparticles
bility and dispersibility in deionized water. The crystal structure The aim of this study was to evaluate and compare the effect on
of pSiNRs@SPIONs was investigated by powder X-ray diffraction mitochondrial activity of the formulations. To evaluate the
(PXRD) technique and displayed typical peaks that correspond viability of F98 rat GBM cells, their mitochondrial activity was
to the SPIONs (Fig. S19, ESI†). As mentioned previously, in this evaluated in presence of pSiNRs, pSiNRs@SPIONs,
research study, an additional functionality to the nanosystem pSiNRs@SPIONs-BIOT-NFL, or pSiNRs@SPIONs-FAM-NFL at
has been added by the loading of uorescent dye (DiD), which different concentrations (25, 50, 100 and 200 mg mL
1) for 72
was realized on pSiNRs for pSiNRs-DiD (Scheme S3, ESI†) and hours. Cells were also treated with 1 mg mL
1 of colchicine, used
simultaneously with the SPIONs for pSiNRs-DiD@SPIONs as a positive control, because colchicine interacts with tubulin
(Scheme S4, ESI†). Several characterizations techniques and disrupts the assembly of microtubules.49 In Fig. 4, MTS
including DLS, ZP, FTIR, and UV-vis spectroscopy conrmed the assay showed a strong decrease of F98 cells viability in presence
successful loading of the DiD dye into the mesoporous structure of colchicine. However, no cellular toxicity of pSiNRs and

© 2022 The Author(s). Published by the Royal Society of Chemistry RSC Adv., 2022, 12, 11708–11714 | 11711
RSC Advances Paper

Fig. 4 In vitro effects of the pSiNRs without or with SPIONs, and

without or with NFL-peptides (BIOT or FAM) on mitochondrial activity
of rat glioblastoma cells (F98). Cells were treated with the formulations
(25, 50, 100 or 200 mg mL
1) or with the positive control colchicine
(col, 1 mg mL
1), for 72 hours, and mitochondrial activity was evaluated
by MTS assay. Experiments were performed at least in triplicate. Data
are represented as mean  SEM. Statistical analysis was performed
with Student's t-test (**p < 0.005 and ***p < 0.001).

pSiNRs@SPIONs, even at the highest concentrations, was

observed. When the NFL-peptide (BIOT or FAM) was added on
pSiNRs@SPIONs, a toxicity was observed with the highest
Fig. 5 Biological transmission electronic microscopy images illus-
concentration (200 mg mL
1). The BIOT-NFL-peptide seems to trating cellular internalization of pSiNRs@SPIONs (a), pSiNRs@SPIONs-
be more effective than the FAM-NFL-peptide. The same experi- BIOT-NFL (b), or pSiNRs@SPIONs-FAM-NFL (c) in rat glioblastoma
ments were carried out on neuroblastoma cells (SH-SY5Y), and cells. F98 cells were incubated 72 hours at 37  C with nanoparticles at
no effect on mitochondrial activity was observed (Fig. S34, ESI†). 200 mg mL
1. Images were taken with a TEM. Scale bars: 1 mm (left
images) and 0.5 mm (right images). N for nucleus and V for vacuoles.
These results demonstrate that the NFL-peptide retains its
biological activity on GBM cells.

DiD@SPIONs-BIOT-NFL, and pSiNRs-DiD@SPIONs-FAM-NFL)

Cellular internalization of nanoparticles by transmission at 10 mg mL
1 were tested on F98 cells for 24 hours. The
electronic microscopy concentration of 10 mg mL
1 was chosen for confocal experi-
In the study, we evaluated the capacity of pSiNRs@SPIONs ment to prevent any signal saturation of DiD. Orthogonal
coupled or not with the NFL-peptide (BIOT or FAM) to be projections obtained with confocal microscope images are
internalized in GBM cells. F98 cells were treated with presented in the Fig. 6. In the Fig. 6, we observed a DiD uo-
pSiNRs@SPIONS, pSiNRs@SPIONs-BIOT-NFL, or rescence in the GBM cells with all conditions demonstrating an
pSiNRs@SPIONs-FAM-NFL at 200 mg mL
1 for 72 hours. Aer internalization of the nanoparticles in F98 cells. An important
xation and inclusion in the resin, cells were sectioned and variation of DiD uorescence was observed, even though the
observed with a TEM. A treatment with pSiNRs@SPIONs cells were treated under the same protocol. When F98 cells are
showed some nanoparticles are present in cells (Fig. 5a). When treated with pSiNRs-DiD, some cells do not show DiD uores-
the NFL-peptide are added to pSiNRs@SPIONs, the cellular cence. When SPIONs are added to the pSiNRs, the DiD uo-
morphology changed, cells present more vacuoles (V), and more rescence is stronger (Fig. 6) and more surprisingly a green
nanoparticles are detected in the GBM cells (Fig. 5b and c). No uorescence appears (Fig. S35†).
difference is observed between pSiNRs@SPIONs-BIOT-NFL The addition of the NFL-peptide to the pSiNRs-DiD increase
(Fig. 5b) and pSiNRs@SPIONs-FAM-NFL (Fig. 5c). The pres- the DiD uorescence. The confocal observations have demon-
ence of pSiNRs@SPIONs does not affect the biological activity of strated the capacity of pSiNRs-DiD@SPIONs with the NFL-
the NFL-peptide. peptides (BIOT or FAM) to internalize in the GBM cells.
However, the presence of SPIONs combined with pSiNRs
induced a green and a red uorescence. As the green uores-
Cellular internalization of nanoparticles by confocal cence appears when the SPIONs are added, to go further,
microscopy another type of nanoparticles was tested: lipid nanocapsules
To evaluate the cellular internalization of the nanoparticles, (LNC) loaded with DiD, without and with SPIONs (LNC-DiD-
cells were treated with formulations loaded with DiD and the SPIONs). Rat GBM cells were treated with LNC-DiD or with
images were taken with confocal microscope. Four formula- LNC-DiD-SPIONs at 2 mg mL
1 for 24 hours, then cells were
tions (pSiNRs-DiD, pSiNRs-DiD@SPIONs, pSiNRs-

11712 | RSC Adv., 2022, 12, 11708–11714 © 2022 The Author(s). Published by the Royal Society of Chemistry
Paper RSC Advances

effects were demonstrated in particular with the targeted

nanoparticles. The obtained results are promising for the
improvement of GBM therapy, and the work is in progress to
further extend the research on in vivo treatment.

Author contributions
A. C. synthesized and characterized the nanoparticles formu-
lations. A. G. performed and analyzed in vitro experiments. G. G.
and J. E. conceived the idea and led the project. A. C., and A. G.,
prepared the manuscript, and all authors contributed to the
nal version.

Conflicts of interest
The authors declare that they have no conict of interest, with
respect to the research, authorship, and/or publication of this
article.

Acknowledgements
Fig. 6 Confocal experiments show cellular uptake of pSiNRs alone or
functionalized with SPIONs, and with the NFL-peptide (BIOT or FAM) in This project has been supported by ‘Plan Cancer Inserm’. The
rat glioblastoma cells. F98 cells were incubated 24 hours at 37  C with authors gratefully thank to Daniela Neacsa for the XRD
nanoparticles at 10 mg mL
1. Images were taken with a confocal measurements, Carine Maaliki for the TGA measurements,
microscope, nanoparticles loaded with DiD were visualized in red, and
nucleus in blue. Pictures illustrating the orthogonal projections of cells
Rodolphe Perrot from SCIAM (Service Commun d'Imageries et
treated. Experiments were performed at least triplicate. Scale bars: 20 d'Analyses Microscopiques, Angers) for the confocal imaging,
mm. and Florence Manero from SCIAM for electron microscopy.

observed with confocal microscope. Orthogonal projections

Notes and references
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11714 | RSC Adv., 2022, 12, 11708–11714 © 2022 The Author(s). Published by the Royal Society of Chemistry