Abstract:AIM: To investigate the effect of zebularine(Zeb)loaded Poly(ethylene glycol)-block-poly(ε-caprolactone)methyl ether(MePEG-PCL)nanoparticles(NPs)on the viability, attachment, and apoptosis of in vitro cultured lens epithelial cells(LECs).
METHODS: In vitro cultured infant human lens tissue HLE B-3 immortalized cells were distributed randomly divided into six groups. Each group was administered with free Zeb 50μmol/L(ZebF1 group), 100μmol/L(ZebF2 group), Zeb -loaded MePEG-PCL NPs 50μmol/L(ZebNP1 group), Zeb -loaded MePEG-PCL NPs 100μmol/L(ZebNP2 group), MePEG-PCL empty NPs(NPs group)or blank medium(group C)respectively. A tetrazolium dye assay(MTT)test and modified MTT test were performed to determine cell viability and cell attachment. DNA ladder was used to detect the cell apoptosis.
RESULTS: Determined by MTT colorimetric method: Cell proliferation rate of LECs were suppressed by all Zeb administration groups in a concentration-time dependent manner(P<0.05). Compared with the free Zeb groups, the viability of LECs were suppressed more effectively by the same dose of Zeb loaded MePEG-PCL NPs after 24, 48, 96h(P<0.05). Determined by improved MTT colorimetric method: The attachment of LECs were decreased in all Zeb administration groups, Zeb loaded MePEG-PCL NPs had better effect on suppressing the attachment of LECs than the free Zeb groups with same dose(P<0.05). The DNA ladder confirmed that after administration of 96h, group C, NPs group and ZebF1 group showed no DNA fragment, however the DNA fragment were performed in ZebF2, ZebNP1, ZebNP2 groups and displays the trend of ZebNP2> ZebNP1>ZebF2(P<0.05).
CONCLUSION: Zeb loaded MePEG-PCL NPs had better effect on suppressing the viability and attachment of in vitro cultured LECs than the free Zeb groups, as well as enhancing the apoptosis.