Bio-multifunctional noncovalent porphyrin functionalized carbon-based nanocomposite

Abstract

Herein, in a one-pot method, the reduced graphene oxide layers with the assistance of multiwalled carbon nanotubes were decorated to provide a suitable space for the in situ growth of CoNi₂S₄, and the porphyrins were incorporated into the layers as well to increase the sensitivity of the prepared nanostructure. The prepared nanocomposite can establish π-π interactions between the genetic material and on the surface of porphyrin rings. Also, hydrogen bonds between genetic domains and the porphyrin' nitrogen and the surface hydroxyl groups are probable. Furthermore, the potential donor-acceptor relationship between the d⁷ transition metal, cobalt, and the genetic material provides a suitable way to increase the interaction and gene loading , and transfections. The reason for this phenomenon was optimized to increase the EGFP by up to 17.9%. Furthermore, the sensing ability of the nanocomposite towards H₂O₂ was investigated. In this regard, the limit of detection of the H₂O₂ obtained 10 µM. Also, the in situ biosensing ability in the HEK-293 and PC12 cell lines was evaluated by the addition of PMA. The nanocomposite showed the ability to detect the released H₂O₂ after adding the minimum amount of 120 ng/mL of the PMA.

Figure 1

The schematic illustration of the synthesized nanosystem.

Figure 2

FTIR (a), XRD (b), FESEM (c–e), AFM (f,g) ,and TEM (h–k) of the synthesized nanocomposite.

Figure 3

Fluorescence emission spectra of the synthesized 3D nanocomposite in the presence of different concentrations of hydrogen peroxide (a), FESEM (b–e), and AFM (f–i) images of the synthesized 3D nanocomposite after exposure to the hydrogen peroxide.

Figure 4

Fluorescence emission spectra of the synthesized 3D nanocomposite in the presence of different concentrations of PMA added to the HEK-293 (a) and PC12 (b) cell lines. 2D (c) and 3D (d) AFM images and (g and h) FESEM images of the synthesized 3D nanocomposite after removing from the PC12 culture and exposure to the hydrogen peroxide; 2D (e) and 3D (f) AFM images and (i and j) FESEM images of the synthesized 3D nanocomposite after removing from the HEK-293 culture and exposure to the hydrogen peroxide.

Figure 5

MTT results of the rGO/MWCNT/CoNi₂S₄, rGO/MWCNT/CoNi₂S₄/H₂TMP, rGO/MWCNT/CoNi₂S₄/H₂TMP + CoNi₂S₄ (a) and rGO/MWCNT/CoNi₂S₄/H₂TMP + CoNi₂S₄ (b) on the (a) HEK-293, (b) HeLa, (c) HepG2 and (d) PC12 cell lines after treatment time of 48 h; and rGO/MWCNT/CoNi₂S₄, rGO/MWCNT/CoNi₂S₄/H₂TMP, rGO/MWCNT/CoNi₂S₄/H₂TMP + CoNi₂S₄ (a) and rGO/MWCNT/CoNi₂S₄/H₂TMP + CoNi₂S₄ (b) on the (e) HEK-293, (f) HeLa, (g) HepG2 and (h) PC12 cell lines after treatment time of 24 h; (a) represents for 10 mg mL⁻¹ and (b) 20 mg mL⁻¹ concentrations of the excess CoNi₂S₄. *p value < 0.05 and **p value < 0.01.

Figure 6

(a) The zeta potential results of the synthesized 3D nanocomposite along with each compartment of that and by addition of the excess amount of CoNi₂S₄ (through blending with the nanocomposite) with (a) 10 mg mL⁻¹ and (b) 20 mg mL⁻¹ concentrations; and (b) the particle size results of the synthesized 3D nanocomposite along with each compartment blended with CRISPR/Cas9 of that and by addition of the excess amount of CoNi₂S₄ (through blending with the nanocomposite) with (a) 10 mg mL⁻¹ and (b) 20 mg Ml^–1 concentrations. One point regarding the zeta potential and particle size is that they were investigated in the presence of CRISPR/Cas9 and its biological matrix.

Figure 7

The results of (a–f) 2D fluorescence microscopy, and (g) GFP positive cells for the synthesized nanocomposite at different WR’s of nanocomposite (nC)/CRISPR/Cas9 (CC) on HEK-293 cell line. The data indicate the 2D fluorescence microscopy and EGFP read are presented as the mean (± SD) from three independent experiments. The Scale bar of (a,b) is 50 µm, and for (c–f) is 10 µm. The normal (h–i) FESEM analysis of the synthesized 3D nanocomposite degraded in the presence of different concentrations of hydrogen peroxide (same as the procedure conducted and shown in Fig. 3), and the normal (j–k) FESEM analysis of the synthesized 3D nanocomposite were removed from the cell culture after the pCRISPR gene transfection experiments.