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Pharmaceutical Care and Research: 2018; 18(1):11-15,20
DOI: 10.5428/pcar20180103
Construction and in vitro evaluation of peptide-modified carboxylated multiwalled carbon nanotube gene vector
1. XIA QingMing1(1.Department of Pharmacy,Changhai Hospital,Second Military Medical University,Shanghai 200433,China
2. GONG ChunAi1(1.Department of Pharmacy,Changhai Hospital,Second Military Medical University,Shanghai 200433,China )
3. GU FenFen2(2.Department of Pharmacy,Xinhua Hospital Affiliated to School of Medicine,Shanghai Jiaotong University,Shanghai 200093,China )
4. HU ChuLing3(3.Department of Pharmacy,Jiaxing Maternity and Child Health Care Hospital,Zhejiang Jiaxing 314050,China )
5. GAO Shen1(1.Department of Pharmacy,Changhai Hospital,Second Military Medical University,Shanghai 200433,China
ABSTRACT  Objective: To prepare peptide modified carboxylated multiwalled carbon nanotubes(MHR) gene vector,and to evaluate its transfection efficiency and cytotoxicity in HEK293 cells.Methods:MHR was obtained by linking the carboxylated multiwalled carbon nanotubes(MWCNTs) with peptide(HR) consisting of arginine(R) and histidine(H) through amide group in a certain ratio of mass.The construction of MHR was characterized by 1H-NMR,infrared spectrum and thermogravimetric analysis.The particle size and zeta potential of purified MHR were determined by laser particle size analyzer.The encapsulation ability of MHR to pEGFP was determined by gel electrophoresis.MHR/pEGFP nanoparticles were co-cultured with HEK293 cells and its cellular uptake and transfection efficiency were investigated.The cytotoxicity of MHR and MWCNTs-COOH to DU145 and RAW264.7 cells was also evaluated.Results: MHR synthesis was confirmed by its structural identification.The cellular uptake and transfection efficiency of HEK 293 cells to MHR/pEGFP at a N/P ratio of 20 was higher than that at other N/P ratios,and MHR/pGL3 compound uptake rate by RAW264.7 cells at a N/P ratio of 20 was about 2.4 folds higher than that of pGL3,and there was statistical significance when comparisons were made between the two (P<0.05).Cell toxicity test showed that the viability of DU145 and RAW264.7 cells treated by MHR for 24 h or 48 h was still more than 80%,even though the concentration of MHR reached 640 μg/ml.However,the viability of the cells decreased markedly when the concentration of MWCNTs-COOH decreased to 320 μg/ml,and when the concentration of MWCNTs-COOH was 640 μg/ml,the viability of the cell was lower than 20%.Conclusion: MHR seems to be a promising efficient gene vector with low cytotoxicity.
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XIA QingMing1,GONG ChunAi1,GU FenFen2,HU ChuLing3,GAO Shen1,. Construction and in vitro evaluation of peptide-modified carboxylated multiwalled carbon nanotube gene vector[J]. Pharmaceutical Care and Research / yao xue fu wu yu yan jiu. 2018; 18(1): 11-15,20.
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