Figure 5
On-site delivery of FTY720 recruits blood-derived non-classical monocytes and increases the frequency of alternatively activated macrophages. PLGA films loaded with FTY720 were implanted immediately after DWC surgery. (a) Proportion of latex bead-positive (LX+) Ly6Clo monocytes that were derived from blood in tissue 3 days post-injury following labeling of Ly6Clo monocytes. (b) Overall frequency of CD206+ F4/80+CD11b+ macrophages in the presence of FTY720. (c) LX+ cells in tissue surrounding unloaded or FTY720-loaded PLGA implants. (d) Proportion of LX+ Ly6Chi monocytes that were derived from blood following labeling of Ly6Chi monocytes with Clod Lip. (e) Overall frequency of CD206+ F4/80+CD11b+ macrophages in the presence of FTY720 after Clod Lip administration. (f) LX+ cells in tissue surrounding unloaded or FTY720-loaded PLGA implants. (g) Ly6C expression in CX3CR1hi monocytes and CX3CR1lo monocytes obtained from the blood of CX3CR1GFP/+ mice identify Ly6Clo monocytes and Ly6Chi monocytes, respectively. (h,i) Surface S1PR3 expression in blood CX3CR1hi cells and CX3CR1lo cells. To control for background staining, CX3CR1GFP/+ cells were stained with secondary antibody (2° Ab) only, which is shown in the gray histogram. Scale bars, 500 µm. Data presented as mean ± S.E.M. Statistical analyses were performed using two-tailed t-tests. *p < 0.05, n = 4–10 animals per group.
