It was reported that the cytotoxicity of beta-catenin mutation PEI-grafted MWNTs is higher than 25-kDa PEI alone in human lung cancer cells (H1299), suggesting that MWNTs enhance the cytotoxicity of PEI . Studies on Daphnia magna also demonstrated that PEI coating increased MWNT
toxicity, which was associated with the size of PEI coating, but not the surface charge of PEI . In contrast, our results suggest that cell viability was higher in the presence of PEI-NH-SWNTs and PEI-NH-MWNTs compared to pure 25-kDa PEI (Figure 9). Liu et al. applied a different Pitavastatin in vitro approach to obtain PEI-grafted MWNTs but reached a similar conclusion to this study by demonstrating that, at concentrations higher than 15 μg/ml, 25-kDa PEI alone is more toxic to 293, HepG2, and COS7 cells compared to PEI-grafted MWNTs . In addition, Wang et al. indicated that PEI-functionalized SWNTs exhibited no significant cytotoxicity to PC-3 cells at concentrations lower than 30 μg/ml but may lead to an increase in apoptosis . In addition to concentration, cytotoxicity of carbon nanotubes
is correlated with the type of functionalization [43, 44], the degree of agglomeration [32, 33], as well as selleck compound nanotube length . Pathways leading to carbon nanotube cytotoxicity were mainly related to DNA damage and the induction of reactive oxygen species . Nevertheless, due to the difference in the types and synthetic procedures of PEI-functionalized carbon nanotubes between this and previous studies and the tolerance of various cells or tissues to the nanomaterial, the cause of carbon nanotube cytotoxicity remains to be investigated. Results from EMSA
showed that at PEI-NH-SWNT/siGAPDH and PEI-NH-MWNT/siGAPDH mass ratios of 80:1 and 160:1, respectively, siGAPDH was completely complexed with PEI-NH-CNTs (Figure 8). However, suppression of GAPDH mRNA expression was observed at relatively lower mass ratios of 1:1 to 1:20 (Figure 10). Such discrepancy in the effective ratios of functionalized carbon nanotubes to siRNAs or DNAs in EMSA and in gene delivery is also presented in previous studies [18, 20, 23]. Amino-functionalized Non-specific serine/threonine protein kinase MWNTs (MWNT-NH3 +) is unable to completely retard the migration of siRNAs in EMSA at a MWNT-NH3 +/siRNA mass ratio of 80:1, but the cationic MWNTs successfully delayed tumor growth in animal models when complexed with siRNAs at a mass ratio of 8:1 . These findings implicate that complete binding of siRNAs by PEI-NH-CNTs may not be necessary for a successful intracellular siRNA delivery. Increasing the amount of PEI-NH-CNTs relative to siRNAs may provide more stable complexes of PEI-NH-CNT/siRNA but may possibly hinder the dissociation of siRNAs from PEI-NH-CNTs once the complex enters the cytosol. Carbon nanotubes are considered an efficient carrier for nonviral gene delivery.