In vivoantitumor activity assessment in localized human NHL xeno-transplant models Daudi cells Selleckchem ZD1839 (1 × 107) in 100 μL of PBS buffer were inoculated subcutaneously into the lateral flank of 6-week-old SCID mice. When the tumors reached about 50 to 60 mm3 in volume, the inoculated mice were randomly assigned to four groups with four each for the treatment of PBS, free ADR, PC-ADR-BSA, and PC-ADR-Fab (with an equivalent amount of 5 mg/kg ADR) via the tail vein weekly for three times. Post-operation monitoring was exercised at least once a day, and the tumor size was measured in two perpendicular diameters
with precision calipers every 3 days and calculated in a range of 60 days. Tumor volume was measured according to the following formula [25]: where length
and width Selleck MK0683 refers to the longest and the shortest diameters of tumors, respectively. Statistical analysis Data were expressed as the means ± standard deviation (SD). Statistical analysis was performed by Student’s t test or one way ANOVA to identify significant differences unless otherwise indicated. Differences were considered significant at a P value of <0.05. Results Characterization of the liposome It has been firmly established that size distribution of a liposome strongly affect its in vitro and in vivo performances [17, 25]. Therefore, we find more firstly assessed the size distribution of our liposome after the successful fabrication. Figure 2A shows the size distribution of irrad and non-irrad liposomes. It was illustrated that an 11% decrease in mean size was occurred after UV irradiation (from approximately 321 nm before irradiation to 285 nm after irradiation). This interesting physical change was validated by morphology analysis using
a TEM, of which the results suggested that both the irrad and non-irrad liposome showed a regular spherical morphology with different diameters (Figure 2B). Figure 2 Properties of CD20 targeting liposomes. (A) Size distribution of liposomes before or after UV irradiation. (B) The TEM morphology of the liposomes before or after UV irradiation, scale bar 0.5 μm. (C) The drug release profile of ADR-loaded liposomes before or after UV irradiation. (D) The cytotoxicity profile of the empty liposomes PC-BSA Decitabine and PC-Fab incubated with CD20 overexpressed Raji cells. Fab fragment loading The number of Fab fragments per liposome was estimated on the basis of Kozlowska’s ideas according to the following equation [35]: Firstly, the liposomal M w was estimated to be 1.22 × 107 g/mol by SLS analysis (Table 1), and the Fab concentration in liposome solution was quantified to be 52.2 μg/mL by determining the A260/A280 by Nano VueTM. Besides, the total mass of liposomes in the suspensions (total volume 2.85 mL) can be calculated from the original polymer amount of 2 mg PC and 0.25 mg Mal-PEG plus the detected amount of Fab (52.2 μg/mL × 2.85 mL) to be approximately 2,398.8 μg. The total mass of liposomes per milliliter can be calculated to be 841.7 μg (2,398.