Abstract
Purpose :
The direct interaction between transient receptor potential vanilloid 1 (TRPV1) channel, inflammation, and cell death remains unclear. In this study, we co-assembled the TRPV1-targeted KFQ12/V1-Cal (KVC) hydrogel to test the hypothesis that decrease of TRPV1 expression and activation could alleviate pyroptosis in dry eye disease.
Methods :
After synthesized of V1-Cal and KFQ12, morphology, structure, mechanical properties, release profile, etc. with (or w/o) V1-Cal were conducted according to previous methods. Hyperosmolarity stressed human corneal epithelial cells and environment-induced murine dry eye model constructed, TRPV1 status, calcium signaling, and pyroptosis pathways were determined by immunofluorescence, Western Blot, qPCR, etc.
Results :
1%, 1.5%, and 2% (w/v) KVC aqueous solutions underwent sol-gel transition immediately after adding to physiological pH buffer, medium, and tear. Circular dichroism (CD) spectra indicated β-sheet secondary structures both in KFQ12 and KVC. Rheological results prompted that gel had a shear thinning and recovery ability when switching of shear strain between 1% and 100% (Fig. B). Cumulative release profile showed that the release rate decreased with increasing concentration (Fig. C). Gel also provided a scaffold for cell crawling in the scratch test. Ocular surface retention tests indicated that 1.5% gel had the longest retention time. Clinical examinations including fluorescein staining, Schirmer tests and TBUT, combined with pathological results showed KVC gel significantly reduced dry eye signs (Fig. D). Other results, obtained from protein and mRNA analysis confirmed that V1-Cal/KVC blocked hyperosmotic stress-induced TRPV1 channel activation, calcium influx and downstream NLRP3-CASP1-GSDMD pyroptosis (Fig. A).
Conclusions :
We synthesized and prepared a co-assembly KVC hydrogel which could significantly prolong drug retention and release time on ocular surface and reduce the eye drops frequency. High water content effectively reduced tear evaporation. β-sheet packing together formed a stable scaffold for epithelium recovery and offered excellent thixotropy to resist shear stress during blink. Vital component V1-Cal effectively alleviated dry eye clinical signs and pyroptosis levels. The study would offer novel insights and treatments of this disturbing disease.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.