Alginate-encapsulated laccase systems for controlled enzyme release: From macroscale carriers to nanoparticles and preliminary<i> in</i><i> vitro</i> evaluation in colorectal cancer cells

Bah M., Bakir E. K., ASABUWA NGWABEBHOH F., YÜZÜGÜLLÜ KARAKUŞ Y.

JOURNAL OF DRUG DELIVERY SCIENCE AND TECHNOLOGY, cilt.127, 2027 (SCI-Expanded, Scopus)

Özet

Colorectal cancer (CRC) remains a major global health burden, highlighting the need for alternative delivery strategies that can improve the stability and controlled release of bioactive agents. In this study, laccase produced from a local Trametes versicolor strain was purified and encapsulated into alginate-based films, macrobeads, and nanoparticles to compare their physicochemical properties, enzyme-release behavior, and preliminary effects on CRC cell viability. Laccase was purified 26-fold with 98% activity recovery through ammonium sulfate precipitation, dialysis, and three-phase partitioning, yielding an approximately 80 kDa protein by SDS-PAGE. Characterization confirmed enzyme incorporation while preserving the characteristic alginate structure. The selected laccase-loaded nanoparticles exhibited a hydrodynamic diameter of 145 f 34.55 nm and the highest encapsulation efficiency (95 f 1.56%), compared with macrobeads (85 f 2.96%) and films (55 f 2.78%). All formulations displayed biphasic release over 240 h, characterized by an initial burst followed by sustained release of catalytically active laccase. Kinetic modeling indicated predominantly Fickian diffusion-controlled release. Preliminary MTT assays demonstrated concentration-dependent reductions in HCT-116 metabolic viability, with comparatively lower effects on HUVEC cells under the tested conditions. Nanoparticles produced the greatest reduction in HCT-116 viability, reaching approximately 33% at 0.5 mg/mL. Overall, alginate-laccase nano-particles demonstrated favorable encapsulation and controlled-release characteristics alongside preliminary antiproliferative effects, supporting further investigation using mechanistic assays, additional CRC and nonmalignant colorectal cell models, and in vivo validation.