Preparation and Release Properties of Helicobacter pylori Recombinant Protein PLGA Microspheres and PLGA-Chitosan Microspheres

CHEN Yu-zuo, LIU Ren-jie, YANG Lu, LI Mao-yuan, LI Wan-yi, WANG Bao-ning

Abstract

To preparethe poly lactic-co-glycolic acid (PLGA) microspheres and PLGA-chitosan microspheres containing Helicobacter pylori recombinant protein, namely the BIB protein, and to explore their optimal preparation parameters and in vitro release performance in gastric and intestinal fluids.  Methods  Double emulsions (water-in-oil-in-water, or W1/O/W2) solvent evaporation method was used to prepare the BIB-PLGA microspheres and the BIB-PLGA-chitosan microspheres. Univariate analysis was done to study the impact of the water-to-oil ratio (W1/O), PLGA mass fraction and PVA concentration on the morphology, particle size, polydispersity index (PDI), encapsulation efficiency (EE), and drug loading (DL) so as to identify the optimal parameters. Bicinchoninic acid (BCA) assay was used to determine the protein concentration and the release efficiency of BIB.  Results   The optimal preparation parameters identified in the study were as follows: W1/O at 1∶2, PLGA mass fraction at 5%, and PVA mass fraction at 0.2%. The BIB-PLGA microspheres were found to be (2.11±0.08) μm in particle size, 0.35±0.18 in PDI, (78.20±1.73)% in EE and (10.58±0.23)% in DL. The BIB-PLGA-chitosan microspheres were (2.28±0.52) μm in particle size, 0.39±0.54 in PDI, and (78.87±1.30)% and (15.50±0.25)% in EE and DL, respectively. Both BIB-PLGA microspheres and BIB-PLGA-chitosan microspheres showed slow-release property in gastric and intestinal fluids in vitro, with BIB-PLGA-chitosan microspheres showing better slow-release performance.  Conclusion  The BIB-PLGA microspheres and BIB-PLGA-chitosan microspheres prepared with the double emulsions solvent evaporation method showed high DL and EE, controllable particle sizes, dispersive appearance, and slow-release property in gastric and intestinal fluids in vitro.

 

Keywords: Helicobacter pylori recombinant protein (BIB protein), Poly lactic-co-glycolic acid microsphere, PLGA-chitosan microsphere

 

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