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Supplementary data

Design and characterisation of novel sorafenib-loaded carbon nanotubes with distinct tumour-suppressive activity in hepatocellular carcinoma

Mahmoud M.A. El-Sayed1, Mahmoud E. Mostafa2, Eman Alaaeldin2,3, Hatem

A. A. Sarhan2, Montaser Sh. A. Shaykoon4, Shady Allam5, Ahmed R.H.

Ahmed6, and Bakheet E. M. Elsadek7,*

1Department of Pharmaceutics and Clinical Pharmacy, Faculty of Pharmacy,

Sohag University, Egypt2Department of Pharmaceutics, Faculty of Pharmacy, Minia University, Egypt3Department of Clinical Pharmacy, Deraya University, Minia, Egypt4Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Al-Azhar

University, Assiut Branch, Egypt5Department of Pharmacology and Toxicology, Faculty of

Pharmacy, Kafrelsheikh University, Egypt6Department of Pathology, Faculty of Medicine, Sohag University, Egypt7Department of Biochemistry and Molecular Biology, Faculty of Pharmacy, Al-

Azhar University, Assiut Branch, Egypt

*Corresponding author: To whom correspondence should be addressed:

Dr. Bakheet E. M. Elsadek, Associate Professor of Biochemistry and

Molecular Biology, Faculty of Pharmacy, Al–Azhar University, Assiut Branch,

P. O. Box 71524, Assiut, Egypt. E–mail: bakheet.elkot@azhar.edu.eg; Tel.:

+201110596270

ORCID: 0000-0001-5444-0245

Keywords: Carbon nanotubes, Sorafenib, DENA, Hepatocellular carcinoma,

Microcapsules

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Zeta potential estimations

Figure 1S. Zeta potential of (A) Pristine CNTs, (B) CNTs-COOH, (C) CNTs-

COCl, (D) CNTs-PEG (E) CNTs-SFN

2

DSC spectroscopy

Figure 2Sa: DSC thermogram of Calcium Chloride

Figure 2Sb: DSC thermogram of, Sodium Alginate

3

Figure 2Sc: DSC thermogram of, Sorafenib tosylate

Figure 2Sd: DSC thermogram of plain microcapsules

4

Figure 2Se: DSC thermogram of MWCNTs-SFN Microencapsulation.

5

Figure 3S: SEM images of the drug-loaded microcapsules after a three

months storage period at 25°C (A), 30°C (B), and 40°C (C) in a RH of

75±5%, at different magnification powers.

6

Morphological changes in the liver features

Figure 4S: Representative photographs of livers from the control group (A1–

5); DENA group (B1–4); SFN group (C1–4); and CNTs-SFN-MCs group

(D1–5) group.

7

Western blot results

Figure 5S: Representative western blot analysis of GP73 (A1-2), GPC-3 (B1-2) and HSP70 (C1-2) expressions in liver tissues of different groups.

8

Immunofluorescence staining

Figure 6S: Representative illustration of liver tissue sections immunofluorescence stained for detection of CTGF, FGF-2, HIF-1α, and mTOR as HCC-relevant biomarkers in untreated control (A), DENA (B), DENA/SFN (C), and DENA/CNTs-SFN-MCs (D) groups.

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Figure 7S: Representative illustration of liver tissue sections immunofluorescence stained for detection of CTGF, FGF-2, HIF-1α, and mTOR as HCC-relevant biomarkers in untreated control (A), DENA (B), DENA/SFN (C), and DENA/CNTs-SFN-MCs (D) groups.

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Histopathological examinations

Figure 8S: Sections of liver tissue obtained from untreated control rats (A), DENA treated rats (B), DENA/SFN treated rats (C) and DENA/ CNTs-SFN-MCs-treated rats (D). Magnification power is x10.

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Figure 9S: Sections of liver tissue obtained from untreated control rats (A), DENA treated rats (B), DENA/SFN treated rats (C) and DENA/CNTs-SFN-MCs-treated rats (D). Magnification power is x20.

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