Supplementary Materials http://advances. GMNP weighed against the NR-2%PEG and the NR-6%PEG. Fig. S7. Mechanistic studies. Fig. S8. Protein corona-related and biodistribution studies. Fig. S9. The molecular models used in the CGMD simulation. Fig. TDZD-8 S10. Diffusion and internalization simulations. Movie S1. Diffusion pattern of GMNP in the mucus detected by STED. Movie S2. Diffusion pattern of SNS in the mucus detected by STED. Movie S3. Diffusion pattern of LNS in the mucus detected by STED. Movie S4. Diffusion pattern of NR-2%PEG in the mucus detected by STED. Movie S5. Diffusion pattern of NR-6%PEG in the mucus detected by STED. Movie S6. Simulation of NR with anisotropic PEG density diffusing in natural hydrogels. Abstract Polyethylene glycols (PEGs) can enhance the diffusivity TDZD-8 of nanoparticles (NPs) in natural hydrogels, while extended PEG stores impede cellular uptake of NPs severely. Influenced by intrusive bacteria with flagellum-driven fimbriae-mediated and mucus-penetrating epithelium-adhering capabilities, we created germ-mimetic NPs (GMNPs) to conquer multiple obstacles in mucosal and tumor cells. In vitro research and computational simulations exposed how the tip-specific prolonged PEG stores on GMNP functioned much like flagella, facilitating GMNP diffusion (up to 83.0-fold faster than their counterparts). In the meantime, the loaded PEG stores for the physiques of GMNP mediated solid adhesive relationships with cells much like the fimbriae, preserving cellular uptake efficiency. The in vivo results proved the superior tumor permeability and improved oral bioavailability provided by the GMNP (21.9-fold over administration of crystalline drugs). These findings offer useful guidelines for the rational design of NPs by manipulating surface polymer conformation to realize multiple functions and to enhance delivery efficacy. INTRODUCTION With the evolution of living organisms, the tunable conformations of molecules play an indispensable role in various biological activities, and a similar principle can be applied in synthetic polymers-decorated systems (penetrate the porous barrier of human mucus layer with the function of these long flagellum (< 0.05 and ***< 0.001, one-way analysis of variance (ANOVA) and MAD-3 Bonferronis test. Since the silica backbone is composed of hydroxyl groups (silanol groups) that are highly associated with surface functionalization (= 4). (D) Uptake by mucus-containing E12 cells that simulated the multiple barriers of intestinal mucosa (= 4). (E) Three-dimensional (3D) view of the transport of NPs across multiple E12 barriers at different times. Scale TDZD-8 bar, 15 m. (F) Different cell entry patterns of NPs. The anisotropic TDZD-8 GMNPs adhered to the membrane via their body side, while the isotropic NRs were wrapped through the tip first. MEM, membrane. Scale bar, 500 nm. (G) Schematic of the superior efficiency of GMNPs over their isotropic counterparts in traversing across multiple barriers. (H) Transportation of GMNPs and their isotropic counterparts with optimized 4% PEGylation in tumor spheroids. Scale bar, 100 m. (I) Quantification of fluorescent coverage of NPs at different depths of spheroids. Representative images are presented. Data are means standard error of mean. ns, no significant difference; **< 0.01 and ***< 0.001, one-way ANOVA and Bonferronis test. To investigate the trends in cellular uptake, we then adopted a mucus-secreting HT29-MTX-E12 (E12) cell monolayer to emulate the intestinal absorption barriers in vivo. The negative impact of PEGylation on cellular uptake was assessed first by removing the mucus layer before incubation. Evidently, PEGylation could substantially impede cellular uptake (Fig. 3C). We noticed that compared with the naked NR, the GMNP with 4% PEGylation showed a nonsignificant 8.7% reduction in the uptake amount, while SNS and LNS with 4% PEGylation suffered a 45.0 and 54.0% decrease in uptake, respectively. Given that the PEG chains surrounding SNS and LNS with 4% PEGylation might be in an intermediate state, the total results recommended that such a conformation could, to a certain degree, decrease the cellular uptake..