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International Journal of Nanomedicine Volume 10, 2015 - Issue
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Original Research

Transactivator of transcription (TAT) peptide– chitosan functionalized multiwalled carbon nanotubes as a potential drug delivery vehicle for cancer therapy

Xia DongLaboratory of Bioengineering, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin Key Laboratory of Biomedical Materials, Tianjin, People’s Republic of China
,
Lanxia LiuLaboratory of Bioengineering, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin Key Laboratory of Biomedical Materials, Tianjin, People’s Republic of China
,
Dunwan ZhuLaboratory of Bioengineering, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin Key Laboratory of Biomedical Materials, Tianjin, People’s Republic of China
,
Hailing ZhangLaboratory of Bioengineering, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin Key Laboratory of Biomedical Materials, Tianjin, People’s Republic of China
&
Xigang LengLaboratory of Bioengineering, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin Key Laboratory of Biomedical Materials, Tianjin, People’s Republic of ChinaCorrespondence[email protected]
Pages 3829-3841 | Published online: 03 Jun 2015

Figures & data

Figure 1 1H-NMR spectra analysis.

Abbreviations: TC, transactivator of transcription–chitosan conjugate; CS, chitosan; MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; 1H-NMR, proton nuclear magnetic resonance.

Figure 1 1H-NMR spectra analysis.Abbreviations: TC, transactivator of transcription–chitosan conjugate; CS, chitosan; MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; 1H-NMR, proton nuclear magnetic resonance.

Figure 2 Raman spectra of the p-MWCNTs, MWCNTs-CS, and MWCNTs-TC.

Abbreviations: ID/IG, peak intensity ratio of D and G band; MWCNTs, multiwalled carbon nanotubes; MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; p-MWCNTs, pristine multiwalled carbon nanotubes.

Figure 2 Raman spectra of the p-MWCNTs, MWCNTs-CS, and MWCNTs-TC.Abbreviations: ID/IG, peak intensity ratio of D and G band; MWCNTs, multiwalled carbon nanotubes; MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; p-MWCNTs, pristine multiwalled carbon nanotubes.

Figure 3 Transmission electron microscopic images of MWCNTs-TC, MWCNTs-CS, and p-MWCNTs after storage at 4°C for 2 months.

Notes: (A) MWCNTs-TC, (B) MWCNTs-CS, and (C) p-MWCNTs. The insets show a photo of MWCNTs-TC, MWCNTs-CS, and p-MWCNTs solution; respectively.

Abbreviations: MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; p-MWCNTs, pristine multiwalled carbon nanotubes.

Figure 3 Transmission electron microscopic images of MWCNTs-TC, MWCNTs-CS, and p-MWCNTs after storage at 4°C for 2 months.Notes: (A) MWCNTs-TC, (B) MWCNTs-CS, and (C) p-MWCNTs. The insets show a photo of MWCNTs-TC, MWCNTs-CS, and p-MWCNTs solution; respectively.Abbreviations: MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; p-MWCNTs, pristine multiwalled carbon nanotubes.

Figure 4 FTIR spectra of MWCNTs-TC, MWCNTs-CS, and p-MWCNTs.

Notes: (A) MWCNTs-TC, (B) MWCNTs-CS, and (C) p-MWCNTs.

Abbreviations: FTIR, Fourier transform infrared spectroscopy; MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; p-MWCNTs, pristine multiwalled carbon nanotubes.

Figure 4 FTIR spectra of MWCNTs-TC, MWCNTs-CS, and p-MWCNTs.Notes: (A) MWCNTs-TC, (B) MWCNTs-CS, and (C) p-MWCNTs.Abbreviations: FTIR, Fourier transform infrared spectroscopy; MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; p-MWCNTs, pristine multiwalled carbon nanotubes.

Figure 5 Analysis of p-MWCNTs, MWCNTs-TC, and MWCNTs-CS by TGA.

Abbreviations: MWCNTs, multiwalled carbon nanotubes; MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; p-MWCNTs, pristine multiwalled carbon nanotubes; TGA, Thermogravimetric Analyzer.

Figure 5 Analysis of p-MWCNTs, MWCNTs-TC, and MWCNTs-CS by TGA.Abbreviations: MWCNTs, multiwalled carbon nanotubes; MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; p-MWCNTs, pristine multiwalled carbon nanotubes; TGA, Thermogravimetric Analyzer.

Figure 6 Zeta potential of MWCNTs-TC, MWCNTs-CS, and p-MWCNTs.

Notes: (A) MWCNTs-TC, (B) MWCNTs-CS, and (C) p-MWCNTs. (D) represents the statistical analysis of Zeta potential for three groups of MWCNTs. *P<0.05 versus p-MWCNTs.

Abbreviations: MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; MWCNTs, multiwalled carbon nanotubes; p-MWCNTs, pristine multiwalled carbon nanotubes.

Figure 6 Zeta potential of MWCNTs-TC, MWCNTs-CS, and p-MWCNTs.Notes: (A) MWCNTs-TC, (B) MWCNTs-CS, and (C) p-MWCNTs. (D) represents the statistical analysis of Zeta potential for three groups of MWCNTs. *P<0.05 versus p-MWCNTs.Abbreviations: MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; MWCNTs, multiwalled carbon nanotubes; p-MWCNTs, pristine multiwalled carbon nanotubes.

Figure 7 Toxicity of MWCNTs-TC or MWCNTs-CS against MD-MBA-231, HUVEC, and L929.

Notes: (A) MD-MBA-231; (B) HUVEC; and (C) L929. *P<0.05.

Abbreviations: MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; MWCNTs, multiwalled carbon nanotubes; HUVEC, human umbilical vein endothelial cell.

Figure 7 Toxicity of MWCNTs-TC or MWCNTs-CS against MD-MBA-231, HUVEC, and L929.Notes: (A) MD-MBA-231; (B) HUVEC; and (C) L929. *P<0.05.Abbreviations: MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; MWCNTs, multiwalled carbon nanotubes; HUVEC, human umbilical vein endothelial cell.

Figure 8 Laser confocal microscopy for MWCNTs-TC and MWCNTs-CS.

Notes: (A and E) Cell nucleus (DAPI); (B and F) MWCNTs (Alexa Fluor® 488); (C and G) microtubules (Alexa Fluor® 555); and (D and H) merge.

Abbreviations: MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; DAPI, 4′,6-diamidino-2-phenylindole dihydrochloride; MWCNTs, multiwalled carbon nanotubes.

Figure 8 Laser confocal microscopy for MWCNTs-TC and MWCNTs-CS.Notes: (A and E) Cell nucleus (DAPI); (B and F) MWCNTs (Alexa Fluor® 488); (C and G) microtubules (Alexa Fluor® 555); and (D and H) merge.Abbreviations: MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; DAPI, 4′,6-diamidino-2-phenylindole dihydrochloride; MWCNTs, multiwalled carbon nanotubes.

Figure 9 Flow cytometric analysis of cellular uptake.

Notes: (A) Cellular uptake rate; (B) mean fluorescent intensity. *P<0.05 versus MWCNTs-CS.

Abbreviations: MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; FITC, fluorescein isothiocyanate; UL, upper left; UR, upper right; LL, lower left; LR, lower right; geo, geometric mean fluorescence intensity; N/A, not applicable.

Figure 9 Flow cytometric analysis of cellular uptake.Notes: (A) Cellular uptake rate; (B) mean fluorescent intensity. *P<0.05 versus MWCNTs-CS.Abbreviations: MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; FITC, fluorescein isothiocyanate; UL, upper left; UR, upper right; LL, lower left; LR, lower right; geo, geometric mean fluorescence intensity; N/A, not applicable.

Figure 10 Living animal imaging on breast cancer-bearing mice administered with MWCNTs-TC, MWCNTs-CS, or NS.

Notes: (A) 24 h; (B) 48 h; and (C) 72 h.

Abbreviations: MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; h, hours; NS, normal saline.

Figure 10 Living animal imaging on breast cancer-bearing mice administered with MWCNTs-TC, MWCNTs-CS, or NS.Notes: (A) 24 h; (B) 48 h; and (C) 72 h.Abbreviations: MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; h, hours; NS, normal saline.

Figure 11 Direct imaging on the excised organs and tissues.

Notes: (A) 24 h; (B) 48 h; and (C) 72 h.

Abbreviations: MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; h, hours.

Figure 11 Direct imaging on the excised organs and tissues.Notes: (A) 24 h; (B) 48 h; and (C) 72 h.Abbreviations: MWCNTs-CS, chitosan-conjugated multiwalled carbon nanotubes; MWCNTs-TC, transactivator of transcription–chitosan-conjugated multiwalled carbon nanotubes; h, hours.

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