Highly open porous biodegradable microcarriers: in vitro cultivation of chondrocytes for injectable delivery

Injectable cell therapy would provide a patient-friendly procedure for treatment of degenerated or wounded tissue. Biodegradable injectable porous microspheres were fabricated to use as dual-purpose microcarriers for cell culture and injectable scaffold for tissue regeneration. Gas foaming in a water-in-oil-in-water double emulsion was performed for fabricating the well-interconnected porous microcarriers using poly(lactic-co-glycolic acid) (PLGA). The gas foaming conditions were finely tuned to control the structural and morphological characteristics. Porous microcarriers with a mean size of approximately 175 microm and an average pore diameter of approximately 29 microm were produced for cell cultivation and injectable delivery. To promote cell seeding, amine-functionalized porous microcarriers were prepared by blending amine-functionalized PLGA with unreacted PLGA. To assess the porous microcarriers for chondrocyte cultivation, bovine articular chondrocytes were seeded and cultured in vitro in spinner flasks for 4 weeks. Visualization and biochemical analyses of the microcarrier-cell constructs were performed to demonstrate cell proliferation and phenotypic expression. Quantification of deoxyribonucleic acid, glycosaminoglycan, and collagen content showed that much greater cell proliferation and expression of cartilage-specific phenotype were observed for cultures in the following order: amine-functionalized porous microcarriers, porous microcarriers, nonporous microcarriers, and monolayer culture.