Enzyme-Functionalized PLGA Nanoparticles with Enhanced Mucus Permeation Rate

The surface functionalization of poly(lactide-co-glycolide) (PLGA) nanoparticles (NPs) with various proteolytic enzymes (i.e., trypsin, papain, bromelain) via a two-step carbodiimide coupling method is presented. Depending on the initial Enzyme:NPs ratio, enzyme loadings up to 4.0 wt.%, 4.4 wt.% and 5.34 wt.% were achieved for trypsin, papain and bromelain, respectively. All three conjugated enzymes partially maintained their enzymatic activity after their coupling reaction with the NPs. NPs functionalized with papain and bromelain exhibited a three-fold higher permeability in porcine intestinal mucus compared to nonfunctionalized NPs whereas those conjugated with trypsin showed an almost two-fold higher permeability value. Measurements of the diffusion rates of intestinal mucin, using a nuclear magnetic resonance (NMR) technique, further confirmed these observations, as the enzyme-functionalized NPs were proven to be capable of disrupting the mucin gel structure. According to the reported results, the coupling of proteolytic enzymes to the PLGA NPs' surface largely increases the NPs mucus permeability, thus making it a potentially important mucus permeation strategy.