Cytotoxicity of PFOB Nanoparticle Coupled with ICAM-1 Antibody on Cardiomyocytes and its Targeted Adhesion to Injured Cardiomyocytes in vitro

WEI Xin, TANG Hong, SHANG Yan-wen, LI Guo-hua, LI Ao, WANG Lian, SONG Hai-bo, FANG Hua

Abstract

To synthesize perfluorooctylbromide (PFOB) nanoparticle coupled with intercellular adhesion molecule-1 (ICAM-1) monoclonal antibody, and to investigate the characteristics of the nonoparticle and its cytotoxcity on and targeted adhesion to injured cardiomocytes in vitro. Methods PFOB nanoparticle (control group) biotinylated PFOB nanoparticle (experimental group) were coupled with biotinylated ICAM-1 antibody. The combination of ICAM-1 antibody and the nanoparticle was detected by immunofluorescent assay. The cytotoxicity of the nanoparticle on rat cardiomyocytes was determined with MTT assay in vitro. The adhesion of the nanoparticle to normal and TNF-α injured cardiomyocytes were observed and semiquantified with optical microscope. Results ICAM-1 antibody was successfully coupled with biotinylated PFOB nanoparticle at a rate around 95%, which showed green fluorescence under the laser Confocal Scanning Microscope, with (385.3±88.9) nm in size, -(60.3±6.11) mV in electric potential, and 7.0×108/mL in concentrations. No fluorescence was observed with the nonoparticle in the control group. No cytotoxicity of the nonoparticle on rat cardiomyocytes was found. There was limited adhesion of the nanoparticle in the control group to cardiomyocytes, normal or injured. A 10-fold increase in adhesion was detected when the nanopaticle in the experimental group was exposed to the injured cardiomyocytes compared with those exposed to the normal cardiomyocytes[(5.1 ±0.22) vs. (0.5±0.3) nanopaticle per cell, P<0.05]. Conclusion ICAM-1 monoclonal antibody is successfully coupled with PFOB nanoparticle, which can effectively bind to the cardiomyocytes overexpressing ICAM-1 without showing ctytotoxicity in vitro.

 

Keywords: PFOB, Cardiomyocyte, Targeting / ICAM-1, Nanometer

 

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References


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