Single-Emulsion P(HB-HV) Microsphere Preparation Tuned by Copolymer Molar Mass and Additive Interaction

Herein, we describe the production of poly(hydroxybutyrate-co-hydroxyvalerate) [P(HB-HV)]-based microspheres containing coumarin-6 (C6) or pyrene (Py) fluorophores as additives and models for hydrophobic and hydrophilic drug encapsulation. Their photophysical and morphological properties, as well as encapsulation efficiencies, are studied as this work aims to describe the influence of additive hydrophobicity/hydrophilicity on microparticle formation. These properties were studied by scanning electron microscopy, fluorescence confocal laser scanning microscopy (FCLSM), and steady-state fluorescence spectroscopy. The results show that the surfactant concentration, polymer molar mass, emulsification stirring rate, and the presence of the fluorophore and its nature are determinants of the P(HB-HV) microsphere properties. Also, encapsulation efficiency is shown to be governed by synergic effects of these parameters on the formation of microspheres. Moreover, size distribution is proved to be strongly influenced by the surfactant poly(vinyl alcohol) content. FCLSM showed that the fluorophores were efficiently encapsulated in P(HB-HV) microspheres at distinct distributions within the copolymer matrix. Surprisingly, nanospheres were observed in the microsphere surface, suggesting that microspheres are formed from nanosphere coalescence.