Base formulations were prepared in bulk with various solvents. Additional excipients were added to base and homogenized before addition of virus. A virus concentration of 1.25 × 1012 vp per film was selected so that subtle changes in infectious titer could be detected with a standard limiting dilution/infectious titer assay and histochemical staining (19). Films were dispensed into 1-ml unit dose molds (or 100-μl unit dose for mouse studies) using an E3 Repeater pipette (Eppendorf, Hauppauge, NY) and dried in 8 hours (1-ml film) and 4 hours (100-μl film) under ambient temperature and pressure (20°C, 1 atm) and aseptic conditions. Once dry, films were reconstituted in and infectious titer assays performed on HeLa cells (ATCC# CCL-2). Percent recovery was calculated as% Recovery=log(infectious titer of t=1)log(infectious titer of t=0)×100where t = 1 is the infectious titer of a film that was reconstituted after drying, and t = 0 is the infectious titer of virus in the same formulation before drying.

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