Multi-layer PLGA-pullulan-PLGA electrospun nanofibers for probiotic delivery

Encapsulation of bacteria into a polymer matrix can potentially enhance the delivery of probiotics. In this study, we present a multilayer electrospun construct to facilitate enhanced delivery of probiotics, where the internal layer is loaded with bacterial cells and the external layers sandwich the internal layer for increased protection and potential mucoadhesion properties. In this proof of concept study, Lactobacillus rhamnosus GG (LGG), which is a robust and well-studied probiotic strain, was encapsulated into pullulan nanofibers, with two electrospun PLGA (Poly-lactic-co-glycolic acid) layers covering it. According to our in vitro study, there was a large decrease in the viability of LGG released from the monolayer sample (LGG:pullulan). However, the multilayer construct maintained excellent viability and an acceptable storage potential. Our in vivo competition study showed that LGG, delivered by multilayer construct, were able to survive intestinal transit and were recovered from all segments of the intestine. Not only did the multilayer construct perform as well as non-encapsulated spray-dried cells in terms of viability and establishment of LGG, but it also showed some cases of increased LGG colonization from fibers in jejunum and cecum compared to spray-dried LGG three days after dosage. We believe, the multilayer construct has the potential to enhance the delivery of the strains that require additional protection, and can benefit from mucus embedment with the help of the covering layers. Particularly, the use of electrospun fibers for the protection of next-generation probiotics sensitive to oxygen and/or gastric conditions could have major commercial interest.