Multivariate Statistical Optimization of a Modified Protocol of the Ionic Polyelectrolyte Pre-Gelation Method to Synthesize Alginate–Chitosan-Based Nanoparticles

Alginate [ALG] and chitosan [CS] are biomaterials of importance in drug-delivery because of their ability to form ionically-crosslinked nanosystems and polyelectrolyte-complexes under mild conditions. Here, a modified ionic-polyelectrolyte-pre-gelation method, with a controlled addition of reagents and sonication throughout the process, is reported to produce ALG¬¬-CS-based NPs. A mathematical study of the effects of the factors with influence in the process on the properties of NPs has been performed using a two-phase-design-of-experiment-based procedure, something never done to our knowledge. The concentration of ALG, CS and CaCl₂ and the ratio ALG:CS have significant influence on polydispersity (PDI), surface-charge (ZP) and encapsulation efficiency (EE%) of NPs. Moreover, CS flow rate has a significant effect over PDI and EE%. Thus, the values of ALG, CS and CaCl₂ concentration (mg/mL), ALG:CS (mL:mL) and CS flow rate (mL/min) to obtain the minimum-expected PDI (0.168) or the optimized EE% (86.7) are {0.30, 0.79, 1.00, 2.50:1.00, 0.86} or {0.30, 1.00, 1.00, 2.50:1.00, 2.00}, with ALG:CaCl₂ (mL:mL) and CaCl₂ flow rate (mL/min) fixed at 2.50:0.31 and 1.25. Although most of the conditions yielded highly-negative particles (minimum-expected of -67.8 mV using 0.69, 0.30 and 0.13 mg/mL of ALG, CS and CaCl₂ and ALG:CS of 2.50:0.59 mL:mL), varying the mass ratio of ALG:CS allowed for the generation of positively-charged particles (up to +21.1 mV with 0.30, 1.00 and 0.61 mg/mL of ALG, CS and CaCl₂ and ALG:CS of 2.50:1.00 mL:mL). In both cases, ALG:CaCl₂ (mL:mL), CaCl₂ and CS flow rates (mL/min) were fixed at 2.50:0.31, 1.25 and 1.25. The model for the NPs size depends only on CS and CaCl₂ concentrations, with the minimum- or maximum-expected (160 or 635 nm) at 0.30 and 1.00 or 1.00 and 0.30 of CS and CaCl₂, although the method allows a wide range of sizes (144.0-1965.0 nm).