3.3.9. Encapsulation efficiency of drug loaded NPs
The encapsulation efficiency (EE) was determined by weighing the amount of unloaded drug recovered from lyophilization of the supernatant obtained after centrifugation of NPs. The drug encapsulation efficiency was calculated according to the following equation:
Encapsulation efficiency (%) = (Winitial/Wfree)/Winitial *100%.
Where, Winitial and Wfree are the weight of initial drug and the weight of free drug determined in the supernatant after centrifugation, and lyophilization respectively. Each sample was determined in duplicate.
3.3.10. Determination of micelle size and zeta potential
The pellets were resuspended in phosphate buffered saline. The size and zeta potential of blank NPs and …show more content…
CCL-119), human ovarian carcinoma (SK-OV-3, ATCC no. HTB- 77), and human prostate cancer cell line (DU-145, ATCC no. HTB-81) were obtained from American Type Culture Collection. CCRF-CEM cells were grown on 75 cm2 cell culture flasks with Roswell Park Memorial Institute (RPMI)-16 medium. Eagle’s minimum essential medium (EMEM) was used for SK-OV-3 and DU-145 cells, supplemented with 10% fetal bovine serum (FBS), and 1% penicillin−streptomycin solution (10,000 units of penicillin and 10 mg of streptomycin in 0.9% NaCl) in a humidified atmosphere of 5% CO2, 95% air at 37 °C. All bioassays were performed in …show more content…
The results were compared with that of free PTX by MTS assay using CellTiter 96 Aqueous One Solution Cell Proliferation Assay Kit (Promega of USA). All cells were plated overnight in 96 well plates with a density of 5,000 cells per well in 100 µL appropriate growth media overnight in a 96 well plate. The cytotoxic drugs (Paclitaxel, ACIN, AMIN), ACIN/COS NPs, AMIN /COS NPs, ACIN/COS-SMCCGGRGDSK NPs, and AMIN/COS-SMCC-GGRGDSK NPs were added to the wells at drug concentration equivalent to 30 µM for the free drug or the drug loaded NPs. The cells were incubated at 37 oC with 5% carbon dioxide for 96 h. Then 20 µL of MTS reagent was added to each of the 96 well plates. The CCRF-CEM cells were incubated for 3 h, and DU-145 cells were incubated for 1 h at 37 oC with 5% carbon dioxide. The absorbance of the formazan product was measured at 490 nm using microplate reader. The percentage of cytotoxicity was calculated as [(OD value of cells treated with the tested compound) − (OD value of culture medium)]/[(OD value of control cells) − (OD value of culture medium)] ×
The encapsulation efficiency (EE) was determined by weighing the amount of unloaded drug recovered from lyophilization of the supernatant obtained after centrifugation of NPs. The drug encapsulation efficiency was calculated according to the following equation:
Encapsulation efficiency (%) = (Winitial/Wfree)/Winitial *100%.
Where, Winitial and Wfree are the weight of initial drug and the weight of free drug determined in the supernatant after centrifugation, and lyophilization respectively. Each sample was determined in duplicate.
3.3.10. Determination of micelle size and zeta potential
The pellets were resuspended in phosphate buffered saline. The size and zeta potential of blank NPs and …show more content…
CCL-119), human ovarian carcinoma (SK-OV-3, ATCC no. HTB- 77), and human prostate cancer cell line (DU-145, ATCC no. HTB-81) were obtained from American Type Culture Collection. CCRF-CEM cells were grown on 75 cm2 cell culture flasks with Roswell Park Memorial Institute (RPMI)-16 medium. Eagle’s minimum essential medium (EMEM) was used for SK-OV-3 and DU-145 cells, supplemented with 10% fetal bovine serum (FBS), and 1% penicillin−streptomycin solution (10,000 units of penicillin and 10 mg of streptomycin in 0.9% NaCl) in a humidified atmosphere of 5% CO2, 95% air at 37 °C. All bioassays were performed in …show more content…
The results were compared with that of free PTX by MTS assay using CellTiter 96 Aqueous One Solution Cell Proliferation Assay Kit (Promega of USA). All cells were plated overnight in 96 well plates with a density of 5,000 cells per well in 100 µL appropriate growth media overnight in a 96 well plate. The cytotoxic drugs (Paclitaxel, ACIN, AMIN), ACIN/COS NPs, AMIN /COS NPs, ACIN/COS-SMCCGGRGDSK NPs, and AMIN/COS-SMCC-GGRGDSK NPs were added to the wells at drug concentration equivalent to 30 µM for the free drug or the drug loaded NPs. The cells were incubated at 37 oC with 5% carbon dioxide for 96 h. Then 20 µL of MTS reagent was added to each of the 96 well plates. The CCRF-CEM cells were incubated for 3 h, and DU-145 cells were incubated for 1 h at 37 oC with 5% carbon dioxide. The absorbance of the formazan product was measured at 490 nm using microplate reader. The percentage of cytotoxicity was calculated as [(OD value of cells treated with the tested compound) − (OD value of culture medium)]/[(OD value of control cells) − (OD value of culture medium)] ×