Formulation and Evaluation of Oral Anti-IL-1β IgY-Loaded Nanoliposomes in DSS-Induced Colitis

Abstract

Purpose

To develop an oral nanoliposomal formulation of anti-interleukin-1β (IL-1β) immunoglobulin Y (IgY) and evaluate its physicochemical properties, gastric stability, and activity in dextran sulfate sodium (DSS)-induced colitis.

Methods

Recombinant canine IL-1β was expressed and used to immunize hens. Purified IgY was loaded into 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC)/cholesterol nanoliposomes by thin-film hydration. Encapsulation efficiency, particle characteristics, morphology, and residual IgY antigen-binding activity in simulated gastric fluid were assessed. Male Institute of Cancer Research (ICR) mice with DSS-induced colitis received free IgY, empty liposomes, or IgY-loaded nanoliposomes. Disease activity, organ indices, colonic cytokines, histopathology, and Occludin and ZO-1 expression were evaluated.

Results

The IgY showed a maximum endpoint titer of 1:409,600, recognized recombinant canine IL-1β, and showed detectable cross-reactivity with recombinant murine IL-1β by Western blotting. Nanoliposomes achieved approximately 56% encapsulation efficiency, a polydispersity index of approximately 0.35, negative ζ-potential, and intact vesicular morphology. At 150 min in simulated gastric fluid, two parallel samples from the same liposomal preparation retained 43.96% and 35.98% antigen-binding activity, respectively. In DSS-treated mice, the formulation was associated with improved clinical indices, lower colonic IL-1β, IL-6, and TNF-α levels, less histopathological injury, and recovery of barrier-protein staining.

Conclusion

Nanoliposomal encapsulation improved the in vitro gastric stability of anti-IL-1β IgY and was associated with anti-inflammatory and barrier-protective effects in DSS-induced experimental colitis. Western blotting demonstrated detectable cross-reactivity of the anti-canine IL-1β IgY with recombinant murine IL-1β; however, functional neutralization of murine IL-1β was not directly assessed.

~ 100 Word Summary Anti-IL-1β IgY

is a potential oral antibody strategy for intestinal inflammation, but gastric instability limits its use. In this study, IgY raised against recombinant canine IL-1β was incorporated into DSPC/cholesterol nanoliposomes and evaluated for physicochemical properties, simulated gastric stability, and activity in DSS-induced colitis. The formulation achieved approximately 56% encapsulation efficiency and retained substantially more IgY antigen-binding activity than free IgY during gastric exposure. In mice, IgY-loaded nanoliposomes were associated with improved disease indices, reduced colonic cytokines, less histopathological injury, and recovery of Occludin and ZO-1 staining. These findings support nanoliposomal protection of IgY during oral delivery, and detectable cross-reactivity with recombinant murine IL-1β was further confirmed by Western blotting.

25–30 Word Teaser

Nanoliposomal encapsulation improved gastric stability of anti-IL-1β IgY and was associated with reduced inflammation and barrier injury in DSS-induced colitis, supporting further evaluation of oral IgY delivery for intestinal inflammation.