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Sealed Bacterial Ghosts—Novel Targeting Vehicles for Advanced Drug Delivery of Water-soluble Substances

The purpose of the present study was to develop a drug delivery model for water soluble drug substances using the bacterial ghost platform technology. Bacterial ghosts are non-denatured bacterial cell envelopes that are produced by the plasmid encoded gene E mediated lysis. We present a novel method to fill and seal bacterial ghosts for the application as a drug delivery system for fluid, non-anchored substances. E. coli ghosts were filled with the reporter substance calcein and sealed by fusion with membrane vesicles. By flow cytometry and fluorescence microscopy it was shown that bacterial ghosts can be filled with calcein, and that the bacterial ghosts can be sealed by restoring the membranes integrity. The adherence and uptake studies showed that almost all murine macrophages and a lower proportion of human colorectal adenocarcinoma cells took up fluorescence labeled bacterial ghosts. Moreover, these cells also took up effectively sealed E. coli ghosts filled with calcein, which then was released within the cells. Therefore, we propose bacterial ghosts as alternative drug delivery and release vehicles for advanced cell targeting.