Fabrication of chitosan microspheres using vanillin/TPP dual crosslinkers for protein antigens encapsulation

Microspheres were prepared from water soluble chitosan using dual vanillin/TPP crosslinkers. Placebo (C1), Bovine serum albumin (BSA) (C2), monovalent tetanus toxoid (TT) (C3) and divalent tetanus (TT) and diphtheria toxoids (DT) (C4) encapsulated microspheres were studied in terms of size (1-4 μm), encapsulation efficiency (75-80%), swelling and mucoadhesion (56-68%). FT-IR, TGA, XRD and SEM characterization of microspheres suggested specific interaction, more thermal stability, amorphous nature and rough surfaces of encapsulated microspheres. EDS confirmed the co-crosslinking and ninhydrin tests were showing higher crosslinking density. Zeta potential was 47.7 to 66.2 +mV indicating the potential stability of the colloidal system. Equilibrium adsorption isotherms described encapsulated microspheres followed the Langmuir isotherm model, suggesting monolayer adsorption of the mucin on microspheres. In-vitro release studies up to four weeks indicated zero order kinetics and obeyed swelling-controlled super case II transport release mechanism. Thus, the present study could be helpful in developing the multivalent oral vaccine.