Upcoming challenge for oral insulin: formulation and characterization of chitosan- reinforced

insulin alginate nanoparticles

C. Reis1, A. Ribeiro2, S. Vilela1, F. Veiga1
1
Pharmaceutics, Faculty of Pharmacy of Coimbra, Portugal, 2Pharmaceutics, ISCSN - Porto, Portugal
Purpose.
To produce chitosan-coated alginate nanoparticles by emulsification/internal gelation, using insulin as a model protein, and to
characterize them on the following parameters: size distribution, interactions between alginate and chitosan, zeta potential
and encapsulation efficiency of insulin.
Methods.
Alginate nanoparticles were prepared by an emulsification-based method using an internal calcium source. The resultant
nanoparticles were recovered using organic solvents and ultracentrifugation. Chitosan-coating was applied by a two-stage
method. Size distribution analysis of hydrated uncoated and coated nanoparticles was performed by laser diffraction
spectroscopy (LDS). The interactions between the two polymers were assessed by differential scanning calorimetry (DSC)
and Fourier-transform infrared spectrometry (FT-IR). The presence of chitosan on surface of alginate nanoparticles was
assessed by zeta potential. At last, insulin encapsulation efficiency of coated and uncoated alginate nanoparticles was
evaluated direct (by insulin release from precise amount of nanoparticles) and indirectly (by insulin losses during the process)
using Bradford method.
Results.
Chitosan-coating led to an increase on mean diameter of alginate nanoparticles from 1.58 to 18.11 µm. The inclusion of
chitosan led to significant change on surface charge of alginate nanoparticles (-16.3 mV to +14.55 mV). FT-IR as well DSC
data suggested the formation of polyelectrolyte complex due to electrostatic interactions between carboxylate groups of
alginate and ammonium groups of chitosan. Finally, coated nanoparticles provided similar encapsulation efficiency compared
to uncoated nanoparticles with values around 70%.
Conclusion.
Chitosan-reinforced insulin alginate nanoparticles were successfully prepared by creating the appropriate conditions. These
nanoparticles produced by emulsification/internal gelation have potential as carriers for oral delivery of insulin.