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Tumor Discovery Dual-targeting of cancer receptors by nanoparticles

20 s, and 72°C for 20 s, and finally melt curve stage at 95°C A B
for 15 s, 60°C for 60 s, and 95°C for 15 s. Each sample was
assayed in duplicate, and the average cycle threshold (CT)
value was normalized to the housekeeping gene GAPDH
and the fold change expression was calculated by ΔΔCT
method .
[26]
3. Results and discussion

3.1. Characterization of Fe O -DPN-HA-FA
3 4
As described in our previous work, the surface of Fe O
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was modified using the dopamine moiety of “DPN.” Figure 2. FESEM images of (A) Fe O -DPN and (B) Fe O -DPN-HA-FA
Dopamine, which acts as an anchoring agent, could replace under 500 nm magnifications. The average size of Fe O -DPN was 13
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4
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the oleylamine on the surface of Fe O NPs . The final (±0.4) nm, while the average size of Fe O -DPN-HA-FA was 153 (±3.3)
[27]
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size of Fe O -DPN was 13 (±0.4) nm . Additional coating nm because folic acid and hyaluronic acid were conjugated to the surface
[28]
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layers (e.g., polyethylene glycol) were applied to prevent of MNPs.
the agglomeration of MNPs and to conjugate targeting
moieties through covalent bonding. uniform morphology of Fe O -DPN through an average
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Two favored candidates for active targeting of cancer size of 13 (±0.4) nm, in which the DPN group is well
cells are HA and FA, which have high affinity to the CD44 immobilized on the Fe O NPs. Furthermore, as shown
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and folate receptors, respectively. For this purpose, in in Figure 2B, the average size of the Fe O -DPN-
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two separate steps, the carboxyl groups of HA and FA HA-FA was increased after anchoring modifier groups
were activated utilizing molar excess of DCC and NHS to about 153 (±3.3) nm under 75k×. The size change
to react with free amine groups (−NH ). In one of them, of Fe O -DPN-HA-FA revealed in the FESEM images
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activated HA was conjugated to the amine end groups on well proves that surface modifications and conjugation
Fe O -DPN, and in another one, activated FA was linked of anionic polysaccharides, such as HA, significantly
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to added amines (EDA) on HA. All variations in the total increased the diameter of Fe O -DPN. Furthermore, the
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composition of synthesized MNPs were confirmed by presence of HA on the surface of Fe O -DPN improves
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FTIR spectroscopy (Figure S1). the circulation time and bioavailability on exposure to a
biological environment [29] .
All FTIR spectra display the absorption bands at 460,
580, and 640 cm , which belong to the Fe-O stretching 3.2. Loading capacities and release amounts
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vibrations. The absorption peak at 1487 cm indicates the TMX, the best clinical choice for breast cancer treatment,
presence of –NH at the end of Fe O -DPN (A at Figure S1), can trigger major dose-dependent side effects, which
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but it disappeared in the spectrum of Fe O -DPN-HA due are related to its physicochemical properties like poor
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to the amide bond created between carboxyl groups of HA solubility . Hence, the use of lower TMX doses utilizing
[18]
and amine group of Fe O -DPN. Two strong peaks at 1627 nano-based drug delivery systems can be beneficial to
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and 1669 cm confirmed the formation of amide groups reducing drug toxicity and providing more efficient
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(B at Figure S1). drug distribution . For nano-carrier systems, drug
[30]
After adding EDA to Fe O -DPN-HA, absorption peaks loading efficiency is a significant parameter, especially
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[31]
at 3317 and 3402 cm confirmed the existence of primary for hydrophobic drugs with common side effects . To
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amine groups on the surface of MNPs (C at Figure S1). reveal the potential of the Fe O -DPN-HA-FA NPs for
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On the other hand, the N-H bending mixed with the the delivery of controlled TMX, the loading capacity and
C-N stretching gives two absorption peaks at 1581 and in vitro release amount were calculated. Due to the large
1327 cm , respectively. size of the HA molecule and a large number of functional
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groups, the loading efficiency of TMX on modified MNPs
Finally, a strong peak at 1589 cm (D at Figure S1) was 73.9%. It is confirmed that modified HA possesses
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was assigned to the aromatic C=C bonds of FA. Besides more benefits, such as increased resistance versus
that, two strong peaks at 1631 and 1678 cm belonged to enzymatic degradation, and can be the ideal platform for
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the new amide bond after conjugation to FA and the free hydrophobic drug encapsulation and drug release . Our
[32]
carboxyl group of FA, respectively. observation showed that 120 h was needed to stop the UV
The morphology and size of the nanostructures absorption changes, and the maximum amount of released
were confirmed using FESEM. Figure 2A shows a TMX after this period was 81%. This proper and sustainable
Volume 1 Issue 1 (2022) 5 https://doi.org/10.36922/td.v1i1.41