Micelles with a molar ratio (CA-PEI) of 1:4 had the maximum doxorubicin release after 6 days. The micelles exhibited a sustained release pattern of doxorubicin, which was characterized by an initial burst release followed by a slow and continuous drug release. In fact, this is a frequent observation for doxorubicin release reported by a number of researchers [25–29]. Doxorubicin is recognized to form a dimer in aqueous selleck compound media due to the chemical reaction between the 30-NH2 group and the C9 α-ketol side chain. Given that the doxorubicin dimer is almost water insoluble and that its azomethine

bond may readily be cleaved to restore the doxorubicin monomer, the later stage of sustained drug release may involve regenerated doxorubicin in addition to the doxorubicin dimer itself [30]. Table 1 DLC and EE of doxorubicin-loaded micelles CA/PEI DLC (% w/w) EE (% w/w) 1:4 0.89 56.52 1:2 0.96 59.44 1:1 1.06 61.31 3:1 1.28 67.57 4:1 1.19 64.22 Figure 8 Doxorubicin release from CA-PEI micelles at pH

7.4. In vitro cell cytotoxicity As shown in Figure 9, the percent inhibition of cancer cells by the doxorubicin-loaded micelles improved from the 1:4 to the 4:1 combinations. Incorporation of doxorubicin into the CA-PEI micelles increased its cytotoxicity toward cancer cells. The half-maximal inhibitory concentration (IC50) values VX-680 supplier for the doxorubicin-loaded micelles were lower than those for free doxorubicin. The lower percentage inhibition and superior IC50 of doxorubicin compared with those of the doxorubicin-loaded

micelles may well be accredited to the formation of aggregates, which deter drug entry into the cells. In addition, doxorubicin could be removed from tumor sites by drug efflux pumps [31]. In contrast, the enhanced cytotoxicity of the doxorubicin-loaded micelles could be explained by the higher permeability and retention of micelles in tumor cells. In addition, increased penetration of the doxorubicin-loaded micelles makes it Florfenicol possible for the drug to be delivered to the site of action, which is located in the nucleus, and therefore gives more time for doxorubicin to interact with its substrate. The increased cytotoxicity observed toward cancer cells could be linked to an increased production of reactive oxygen species and enhanced apoptosis. The ability of CA to modulate the number of aberrant crypt foci by restraining their development and growth and by eliminating a selected population may also contribute to the cytotoxicity of the doxorubicin-loaded micelles [32]. Both free doxorubicin and entrapped doxorubicin caused cell death in a dose-dependent manner. The cytotoxicity of doxorubicin is likely to increase further in vivo due to the enhanced permeation and retention effects of the loaded micelles. These findings imply that the selective uptake of micelles by cancer cells could reduce the toxicity and adverse effects of doxorubicin.