Physical Characterization and In Vivo Study of Ovalbumin Encapsulated In Alginate Microspheres

Hariyadi, D.M., Purwanti, T., Kusumawati, I., Nirmala, R.N., Maindra, H.M.C.

Microspheres as drug delivery systems have been used widely for oral delivery in the pharmaceuticals formulation. Alginates have received much attention as biodegradable polymer for controlled protein delivery. Ovalbumin was used as model antigen. The aim of the present research was to evaluate physical characteristics of unlyophilized and lyophilized ovalbumin-loaded alginate microspheres produced using aerosolisation technique. Crosslinking agent of BaCl₂ and CaCl₂ were studied. Physical characteristics of ovalbumin-loaded alginate microspheres were evaluated in terms of encapsulation efficiency, yield, particle size, surface morphology and protein integrity. In vivo immune response was studied using hemagglutination test by measuring antibody titre. High entrapment efficiency of unlyophilized ovalbumin-loaded alginate microspheres using crosslinker BaCl₂ and CaCl₂ of above 80% and loading of above 65% was produced. Entrapment efficiency of lyophilized ovalbumin-loaded alginate microspheres using 10% lactose lyoprotectant and maltodextrin were 46% and 54% respectively. For protein loading, ovalbumin-loaded alginate microspheres lyophilized using lactose was 28% and using maltodextrin was around 35% respectively. The microspheres size of less than 8μm of all formulas using BaCl₂ and CaCl₂ both lyophilized and unlyophilized were spherical in shapes and formed smooth surface. Ovalbumin maintained its integrity after release from alginate microspheres showing ovalbumin’s 45 kDa molecular weight indicated its stability has been protected by this delivery system after exposure to acid condition. In vivo immune response by hemaglutination test of lyophilized ovalbumin-loaded alginate microspheres with CaCl₂ crosslinker consist of 10% lactose or 10% maltodextrin and unlyophilized microspheres using CaCl₂ crosslinker showed higher antibody titres than ovalbumin control and blank microspheres. Whereas, unlyophilized ovalbumin-alginate microspheres using Ba²⁺ crosslinking agent resulted antibody titre as low as control and blank microspheres. However, No significant effect of immune response was found between microspheres lyophilized with lactose and maltodextrin. In conclusion, ovalbumin-loaded alginate microspheres crosslinked using CaCl₂ are potential as oral delivery systems and results indicated that lyoprotectants were involved in protecting ovalbumin entrapped in alginate microspheres therefore able to deliver antigen to the target site in order to induce antibody response.

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