Bhat M and Hickey AJ Effect of Chloroquine on Phagolysosomal Fusion in Cultured Guinea Pig Alveolar Macrophages: Implications in Drug Delivery AAPS PharmSci 2000; 2 (4) article 34 (https://www.pharmsci.org/scientificjournals/pharmsci/journal/34.html).

Figures and Tables

Figure 1.Log-probability plot of the size analysis of rhodamine-labeled rifampicin polylactic-co-glycolic acid microspheres (Median diameter 0.473 µm [GSD 2.09]).

Figure 2.Representative photomicrograph of rhodamine-labeled rifampicin polylactic-co-glycolic acid microspheres.

Figure 3.Rhodamine release profile from rhodamine-labeled rifampicin polylactic-co-glycolic acid microspheres (n=5).

Panel 1.Photomicrographs of alveololar macrophages containing rhodamine-labeled rifampicin polylactic-co-glycolic acid (RR-PLGA ) microspheres at a) 24 hours, b) 4 days, and c) 6 days. Cells stayed in culture for 7 days and began detaching. Patterns of punctate to diffuse fluorescence was observed over time, indicating the slow degradation of RR-PLGA microspheres.

Panel 2.Photomicrographs of alveolar macrophages with BioParticles and rhodamine-dextran. a) cells depicting pinpoint red lysosomes b) few cells with freshly phagocytosed BioParticles c) cells displaying orange-yellow fluorescence indicating phagolysosomal fusion

Figure 4.Kinetics of phagolysosomal fusion over time in control and treatment groups (n=5). For each timepoint, cells over 10 fields of view at 40X were scored as fusion/non-fusion events depending on visualization of the orange-yellow color formed on fusion between endosomes and lysosomes. The percentage of.phagolysosomal fusion was then obtained by dividing the number of cells by the total number of cells in the field of view.

Figure 5.Scatter patterns for 5 x 5 pixel areas of a, b values of rhodamine-dextra-treated cells alone (red fluorescence) or fluorescein-labeled BioParticles exposed cells alone (green fluorescence). In the Lab scale system, green to red values range from -120 to +120. The cluster of scatter points on the left-hand side of the graph corresponds to a, b values for 5 x 5 pixel areas (at 60X) treated with only fluorescein-labeled BioParticles, while the cluster of points on the right-hand side corresponds to cells treated with only rhodamine-labeled dextran (70 kd).

Figure 6.Scatter patterns for 5 x 5 pixel areas of a,b values of rhodamine-dextra-treated and fluorescein-labeled BioParticles exposed cells. The profile of the a, b values for the 5 x 5 pixel areas of cells visually displaying an orange-yellow color falls in between the values for the corresponding a, b values for 5 x 5 pixel areas for fluorescein-labeled BioParticles only-and rhodamine-dextran only-treated cells displayed in Figure 5 .