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International Journal of Nanomedicine Volume 12, 2017 - Issue
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Original Research

Delivery of sodium morrhuate to hemangioma endothelial cells using immunoliposomes conjugated with anti-VEGFR2/KDR antibody

Xiaoli Li1 Department of Dermatology, The Second Affiliated Hospital, Xi’an Jiaotong University, Xi’an, People’s Republic of China
,
Xiaoyong Ren2 Department of Otolaryngology-Head and Neck Surgery, The Second Affiliated Hospital, Xi’an Jiaotong University, Xi’an, People’s Republic of China
,
Jianmin Liang2 Department of Otolaryngology-Head and Neck Surgery, The Second Affiliated Hospital, Xi’an Jiaotong University, Xi’an, People’s Republic of China
,
Weijun Ma2 Department of Otolaryngology-Head and Neck Surgery, The Second Affiliated Hospital, Xi’an Jiaotong University, Xi’an, People’s Republic of China
,
Zhenghui Wang2 Department of Otolaryngology-Head and Neck Surgery, The Second Affiliated Hospital, Xi’an Jiaotong University, Xi’an, People’s Republic of ChinaCorrespondence[email protected] [email protected]
&
Zhuangqun Yang3 Department of Plastic and Burns Surgery, The First Affiliated Hospital, Xi’an Jiaotong University, Xi’an, People’s Republic of ChinaCorrespondence[email protected] [email protected]
Pages 6963-6972 | Published online: 19 Sep 2017

Figures & data

Figure 1 Preparation of liposomes coupled with anti-VEGF2. (A) Schematic illustration of the coupling of anti-VEGF2 to liposomes via maleimide-thiol reaction. (B) Particle size distribution of liposomes with anti-VEGF2. (C) TEM examination revealed a uniform spherical shape for the liposomes coupled with anti-VEGF2 (magnification ×5,000).

Abbreviations: Diam, diameter; Mal, maleimide; SPDP, N-succinimidyl-3-(2-pyridyldithio)propionate; TEM, transmission electron microscopy; VEGF2, vascular endothelial growth factor 2; WT, wild type.

Figure 1 Preparation of liposomes coupled with anti-VEGF2. (A) Schematic illustration of the coupling of anti-VEGF2 to liposomes via maleimide-thiol reaction. (B) Particle size distribution of liposomes with anti-VEGF2. (C) TEM examination revealed a uniform spherical shape for the liposomes coupled with anti-VEGF2 (magnification ×5,000).Abbreviations: Diam, diameter; Mal, maleimide; SPDP, N-succinimidyl-3-(2-pyridyldithio)propionate; TEM, transmission electron microscopy; VEGF2, vascular endothelial growth factor 2; WT, wild type.

Figure 2 Characterization of HECs. (A) Morphology of HECs. (B) Immunocytochemistry showed that the cell cultures were positive for factor VIII. (C) Reverse transcription PCR analysis demonstrated that the isolated cells expressed factor VIII and VEGF2 mRNA. Bar =50 μm.

Abbreviations: HEC, hemangioma endothelial cell; PCR, polymerase chain reaction; VEGF2, vascular endothelial growth factor 2.

Figure 2 Characterization of HECs. (A) Morphology of HECs. (B) Immunocytochemistry showed that the cell cultures were positive for factor VIII. (C) Reverse transcription PCR analysis demonstrated that the isolated cells expressed factor VIII and VEGF2 mRNA. Bar =50 μm.Abbreviations: HEC, hemangioma endothelial cell; PCR, polymerase chain reaction; VEGF2, vascular endothelial growth factor 2.

Figure 3 Morphological characteristics of HECs (microscope and Giemsa staining). (A) Control group; (B and F) sodium morrhuate group; (C and G) blank liposome group; (D and H) liposomal sodium morrhuate group; (E and I) sodium morrhuate immunoliposome group. Scale bars =50 μm.

Abbreviation: HEC, hemangioma endothelial cell.

Figure 3 Morphological characteristics of HECs (microscope and Giemsa staining). (A) Control group; (B and F) sodium morrhuate group; (C and G) blank liposome group; (D and H) liposomal sodium morrhuate group; (E and I) sodium morrhuate immunoliposome group. Scale bars =50 μm.Abbreviation: HEC, hemangioma endothelial cell.

Figure 4 Confocal microscopy reveals the targeting of sodium morrhuate immunoliposomes to HECs. (A and a) Control group; (B and b) liposomal sodium morrhuate group; (C and c) sodium morrhuate immunoliposomes group. Scale bars =50 μm.

Abbreviation: HEC, hemangioma endothelial cell.

Figure 4 Confocal microscopy reveals the targeting of sodium morrhuate immunoliposomes to HECs. (A and a) Control group; (B and b) liposomal sodium morrhuate group; (C and c) sodium morrhuate immunoliposomes group. Scale bars =50 μm.Abbreviation: HEC, hemangioma endothelial cell.

Figure 5 HECs were treated with indicated agents for different times and tested for viability by MTT assay.

Abbreviations: HEC, hemangioma endothelial cell; MTT, 3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide; OD, optical density; SM, sodium morrhuate.

Figure 5 HECs were treated with indicated agents for different times and tested for viability by MTT assay.Abbreviations: HEC, hemangioma endothelial cell; MTT, 3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide; OD, optical density; SM, sodium morrhuate.

Figure 6 Apoptosis analysis by flow cytometry. (A) Control group; (B) liposomal sodium morrhuate group; (C) sodium morrhuate immunoliposomes group; (D) quantification of apoptosis.

Note: The sample number is show above each analysis.

Abbreviations: Li, liposome; SM, sodium morrhuate.

Figure 6 Apoptosis analysis by flow cytometry. (A) Control group; (B) liposomal sodium morrhuate group; (C) sodium morrhuate immunoliposomes group; (D) quantification of apoptosis.Note: The sample number is show above each analysis.Abbreviations: Li, liposome; SM, sodium morrhuate.

Figure 7 TEM examination of HECs treated with indicated agents. (A and B) Sodium morrhuate group (×10,000); (C) liposomal sodium morrhuate group (×4,000); (D) sodium morrhuate immunoliposome group (×4,000); (E) control group (×4,000).

Abbreviations: HEC, hemangioma endothelial cell; TEM, transmission electron microscopy.

Figure 7 TEM examination of HECs treated with indicated agents. (A and B) Sodium morrhuate group (×10,000); (C) liposomal sodium morrhuate group (×4,000); (D) sodium morrhuate immunoliposome group (×4,000); (E) control group (×4,000).Abbreviations: HEC, hemangioma endothelial cell; TEM, transmission electron microscopy.

Figure 8 Western blot analysis of indicated proteins from different groups. Representative blots of three independent experiments with similar results are shown.

Abbreviations: Li, liposome; SM, sodium morrhuate.

Figure 8 Western blot analysis of indicated proteins from different groups. Representative blots of three independent experiments with similar results are shown.Abbreviations: Li, liposome; SM, sodium morrhuate.

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