Development of alkaline-stable nanoformulation of nisin: special insights through cytotoxic and antibacterial studies.

Abstract

Nisin, a thermostable, approved food preservative, has limited therapeutic applications because of its high pH and proteolytic enzyme instability. The unavailability of a rapid, simple method of detection also restricts the research of nisin. The objective of this study was to adapt the simple, rapid protein estimation method of detection for nisin formulation and to formulate and evaluate site-specific nanoformulation for therapeutic applications, viz. colon cancer, and anti-bacterial action. Three nanoformulations of nisin with chitosan, gellan gum, and dextran (ECN, EGN, and EDN) were prepared and characterized in vitro. Among three, EGN was selected as a good formulation based on its size surface charge, morphology, drug loading, and release characteristics. FT-IR and DSC revealed the interaction pattern and stability nature. The stability of nisin in an alkaline environment was confirmed by CD. Its therapeutic applications were proved by efficiency against colon cancer cells evaluated by MTT assay and AO/EB staining using Caco-2 cell lines. The in situ sol-gel mechanism imparted by gellan gum was proved the sole reason for the stability and activity of nisin in EGN at lower GIT. This was confirmed (using rheometer) by shear-thickening characteristics of formulation EGN in simulated colon fluid. The antibacterial activity against Staphylococcus aureus by disk diffusion method was also performed to confirm the retention of antimicrobial activity of nisin in EGN. Hence, gellan gum-nisin colloidal nanoparticles are found good candidates for drug delivery at lower GIT and stabilizing alkaline food materials.