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. 2024 Oct 5;10(19):e38904.
doi: 10.1016/j.heliyon.2024.e38904. eCollection 2024 Oct 15.

Synthesis and in vitro study of surface-modified and anti-EGFR DNA aptamer -conjugated chitosan nanoparticles as a potential targeted drug delivery system

Maryam Rahmani Kheyrollahi  1 , Javad Mohammadnejad  2 , Akram Eidi  1 , Hanieh Jafary  1
Affiliations

Synthesis and in vitro study of surface-modified and anti-EGFR DNA aptamer -conjugated chitosan nanoparticles as a potential targeted drug delivery system

Maryam Rahmani Kheyrollahi et al. Heliyon. .

Abstract

Nowadays, finding effective approaches for cancer therapy is one of the significant issues related to human health all over the world. Hence, in this research, we designed and synthesized a novel targeted DDS based on surface-modified chitosan (CS) for the effective delivery of noscapine (NO). As the surface of CS nanoparticles was firstly modified with carboxyl groups and followed by covalent conjugation of DNA-aptamer (Ap) as targeting and receptor blocker agent. Secondly, NO, as a chemotherapeutic agent, was loaded into prepared nano-complex and synthetics were effectively characterized via various analytical devices, including FT-IR, 1H NMR, DLS, Zeta potential analyzer, TGA, TEM, and SEM to verify quality and quantity of synthetics. Drug loading was obtained about 25 % and sustained drug release was observed for nano-complex at different pHs. Then, the cell viability assay was performed on MCF-7 (as breast cancer cell) and HFF-1 (as normal cell) cell lines to investigate cancer cell inhibition potency of nano-complex. Cell viability of cancer cells was 19.84 ± 1.87 % (for C-CS-Ap-NO) and 75.43 ± 2.64 % (for C-CS-Ap) after 72 h of treatment with 400 nM concentration. These results have been confirming the excellent potency of synthesized novel nano-complex as practical DDS in cancer therapy.

Keywords: Cancer cell inhibition; Chitosan nanoparticles; Controlled noscapine release; DNA-Aptamer; Targeted drug delivery.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Fig. 1
Fig. 1
The FT-IR spectra of CS and its derivatives. The absorption peaks are presenting the appropriate surface modification, conjugation of Ap to C-CS, and presence of NO drug in nano-complex which confirms the excellent qualification of syntheses. For the CS sample, the peaks were detected at around 3437 cm−1 (–NH2 and –OH groups stretching vibration), 3337 cm−1 (O−H and N−H stretching vibrations), 1151 cm−1 (asymmetric stretching of −C−O−C− bridge), and 1034 cm−1 (C−N bond). Also, the sharp absorption peaks at around 1420 cm−1 to 1495 cm−1 could be due to the formation of amides bending (CH−NH−C=O) between amine groups of Ap and carboxyl groups of C-CS in the structure of Ap-C-CS nano-complex.
Fig. 2
Fig. 2
1H NMR spectra of CS and its derivatives. (A) 1H NMR of C-CS indicates resonance peaks from 3.00 ppm to 5.00 ppm which are assigned to the protons of glucosamine unit and the peaks at around 1.9 ppm and 2.23 ppm are corresponding to the methyl protons of the N-acetyl group in CS. Also, the peak around 1.2 ppm demonstrates the presence of CH3 of the ester group. (B) 1H NMR of C-CS-Ap shows the methyl proton peaks of Ap between 0.8 ppm and 1.96 ppm and as well as ethyl protons from 2.26 ppm to 2.75 ppm due to the presence of Ap in the structure of nano-complex. (C) 1H NMR of C-CS-Ap-NO nano-complex.
Fig. 3
Fig. 3
Zeta potential values of synthetic nano-complexes. The surface charge of C-CS-Ap (A) and C-CS-Ap-NO (B) formulations are at about +27.7 mV and +26.0 mV, respectively.
Fig. 4
Fig. 4
TEM images of CS derivatives, including C-CS-Ap (A) and C-CS-Ap-NO (B). These images are showing the nano-size, spherical, and globular structures with good dispersion for CS derivatives. The SEM images are also presented a suitable globular morphology and appropriate dispersion for C-CS-Ap (C) and C-CS-Ap-NO (D) nano-complexes.
Fig. 5
Fig. 5
TGA analyses of the C-CS-Ap and C-CS-Ap-NO samples. As shown, the TGA curve of C-CS-Ap specimen has two main weight loss phases: 5 % at 110 °C and 28 % at around 250 °C–290 °C. The weight loss curve of the C-CS-Ap-NO sample was earned in the main phase between 185 °C and 287 °C. Based on these two curves, the drug content of nano-complex was calculated at about 25 %.
Fig. 6
Fig. 6
The drug release profile of pure NO, NO of C-CS-Ap-NO, and CS-NO nano-complexes under pH values of 7.4, 6.0, and 5.0. C-CS-Ap-NO sample shows a controlled release manner at pH 7.4 (as a mimic of physiologic medium) than pHs 6.0 and 5.0. The protonation of the CS chains at pH 6.0 and 5.0 is the main reason for the fast drug release rate that is appropriate for the effective delivery of NO to cancer cells since their environments and especially their endosomal organelles have the acidic condition. Besides, C-CS-Ap-NO indicates the controlled drug release manner than CS-NO specimen which could be due to its surface functionalization with Ap and as well as the surface modification.
Fig. 7
Fig. 7
Effects of CS, C-CS, Ap-C-CS, Ap-C-CS-NO, and pure NO nano-complexes on cell viability of MCF-7 and HFF-1 cell lines after 24 h, 48 h, and 72 h of treatment with various concentrations (100 nM. 200 nM, 400 nM) of samples. The excellent cancer cell growth inhibition is exhibited for the Ap-C-CS-NO specimen. On the other hand, low cytotoxicity is attained for C-CS (for cancer cells) and Ap-C-CS (for normal cells) which is indicating excellent biocompatibility, biosafety, and biodegradability of the synthetic nano-complex.
Fig. 7
Fig. 7
Effects of CS, C-CS, Ap-C-CS, Ap-C-CS-NO, and pure NO nano-complexes on cell viability of MCF-7 and HFF-1 cell lines after 24 h, 48 h, and 72 h of treatment with various concentrations (100 nM. 200 nM, 400 nM) of samples. The excellent cancer cell growth inhibition is exhibited for the Ap-C-CS-NO specimen. On the other hand, low cytotoxicity is attained for C-CS (for cancer cells) and Ap-C-CS (for normal cells) which is indicating excellent biocompatibility, biosafety, and biodegradability of the synthetic nano-complex.
Fig. 8
Fig. 8
Cellular uptake of pure CS nanoparticle (B) and Ap-C-CS nano-complex (C) after 5 h of treatment on MCF7 cell line. A graph is control. The fluorescence intensity of MCF-7 cells showed after treatment with pure CS nanoparticles was acquired 1.65 % on cancer cells while it remarkably increased to more than 50 folds (90.5 %) on cancer cells after treatment with Ap-C-CS nanocarrier at the same circumstances.
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References

    1. Siegel R.L., Miller K.D., Jemal A. Cancer statistics. CA Cancer J Clin. 2017;67:7–30. - PubMed
    1. Ferlay I. Soerjomataram, Dikshit R., Eser S., Mathers C., Rebelo M., et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int. J. Cancer. 2015;136:E359–E386. - PubMed
    1. Yousefi M., Narmani A., Jafari S.M. Dendrimers as efficient nanocarriers for the protection and delivery of bioactive phytochemicals. Adv. Colloid Interface Sci. 2020;278 - PubMed
    1. Narmani A., Yavari K., Mohammadnejad J. Imaging, biodistribution and in vitro study of smart 99mTc-PAMAM G4 dendrimer as novel nano-complex. Colloids Surf., B. 2017;159:232–240. - PubMed
    1. Lee S.H., Bajracharya R., Min J.Y., Han J.-W., Park B.J., Han H.-K. Strategic approaches for colon targeted drug delivery: an overview of recent advancements. Pharmaceutics. 2020;12:68. - PMC - PubMed

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