The mean fluorescence intensity of the cells isolated from the peritoneal cavity 20 or 60hrs after the administration was 4.78 or 47.61 per 10000 cells, respectively. The calculated concentration of Hoechst 33342 was 40.1ng/mL after 20hrs or 491.0ng/mL after 60hrs. Figure 8 (a) Fluorescence intensity of U-937 cells was analyzed after
staining with serial concentrations of Hoechst 33342 using FACS Aria II. The segments P1, P2, P3, or P4 correspond to the range of fluorescence intensity at 0, 10, 100, or 1000ng/mL … In the present study we have used Hoechst 33342-incorporated PLGA to identify, isolate, and characterize cells exposed to this Inhibitors,research,lifescience,medical fluorescent dye. The nuclear staining of Hoechst 33342 in vivo is a powerful marker for the isolation of cells from blood, ascites, LY317615 pleural effusions, and even tissues when the tissue dissociation
and cell isolation protocol is established. In addition, we can also collect cells that are negative for fluorescence. Once the various cells have Inhibitors,research,lifescience,medical been isolated, they can be analyzed for cell type and expression of specific molecules such as surface markers that may be important in cell targeting. One major limitation of the present approach is that Hoechst 33342 used as an imitating drug will be different from the actual drug in terms of molecular weight, structure, electrical charge, and/or presence/absence of specificity Inhibitors,research,lifescience,medical for a target molecule. Nonetheless, the present approach is useful for investigating the likely distribution of released Inhibitors,research,lifescience,medical materials from individual PLGA particles in the microenvironment of target tissues. 4. Conclusion The present study successfully demonstrated that Hoechst 33342-incorporated PLGA particles can be used to simulate the drug exposure of cells in Inhibitors,research,lifescience,medical situ. We isolated cells exposed to this fluorescent dye as well as those that were not. These two classes of cells can then be further characterized, especially with regard to the expression of specific molecules that may be important in the targeting mechanism. The present approach may provide
essential information concerning cell targeting in any type of PLGA DDS. Conflict of Interests The authors certify that there is no conflict of interest’s with any financial organization regarding the material discussed in the paper. Acknowledgments The study Cilengitide was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan and a grant from the Intractable Diseases, the Health and Labor, and Labor Sciences Research Grants from the Ministry of Health, Labor and Welfare of Japan. Some of the results were generated by using the facilities of Biomedical Research Core of Tohoku University Graduate School of Medicine. The authors also acknowledge the support of Tohoku University Global COE Program “Global Nano-Biomedical Engineering kinase inhibitor Imatinib Education and Research Network Centre.