Encapsulation of Bacterial Spores in Nanoorganized Polyelectrolyte Shells

Layer-by-layer assembly uses alternating charged layers of polyionic polymers to coat materials sequentially in a sheath of functionalized nanofilms. Bacterial spores were encapsulated in organized ultrathin shells using layer-by-layer assembly in order to assess the biomaterial as a suitable core and determine the physiological effects of the coating. The shells were constructed on Bacillus subtilis spores using biocompatible polymers polyglutamic acid, polylysine, albumin, lysozyme, gelatin A, protamine sulfate, and chondroitin sulfate. The assembly process was monitored by measuring the electrical surface potential (zeta-potential) of the particles at each stage of assembly. Fluorescent laser confocal microscopy and scanning electron microscopy confirmed the formation of uniform coatings on the spores. The coating surface charge and thickness (20-100 nm) could be selectively tuned by using appropriate polymers and the number of bilayers assembled. The effect of each coating type on germination was assessed and compared to native spores. The coated spores were viable, but the kinetics and extent of germination were changed from control spores in all instances. The results and insight gained from the experiments may be used to design various bioinspired systems. The spores can be made dormant for a desired amount of time using the LbL encapsulation technique and can be made active when appropriate.