Abstract

Therapeutic enzymes hold great promise in cancer treatment, yet their clinical application is often limited by instability in physiological environments. This study investigates the application of a silica–chitosan hybrid matrix as a protective and responsive carrier for glucose oxidase (GOx) and asparaginase (ASNase), two enzymes with potential antitumor properties. The encapsulation process was optimized using GOx, achieving an encapsulation efficiency of up to 84.4 % under high chitosan-to-silica conditions and mechanical agitation. Encapsulated enzymes were evaluated for stability and release under different pH levels, temperatures, and culture media. Both GOx and ASNase exhibited enhanced stability and sustained activity at 37 °C and under alkaline conditions, with superior retention and release in cancer cell culture media (GOx: 76.4 %, ASNase: 85.4 %). In simulated gastrointestinal conditions, a controlled release was observed, reaching 80 % for GOx and 74 % for ASNase at colonic pH. These findings demonstrate that the silica–chitosan system effectively protects enzymes from degradation while enabling pH-responsive, sustained release. The versatility and performance of this biohybrid matrix position it as a promising platform for future biomedical applications, particularly in oral delivery strategies targeting the gastrointestinal tract or tumor microenvironments. © 2025