Poly (l-lactide-co-caprolactone) scaffolds enhanced with poly (β-hydroxybutyrate-co-β-hydroxyvalerate) microspheres for cartilage regeneration

Biodegradable polymers, either as porous scaffolds or microspheres, have been investigated broadly for cartilage tissue engineering. A combination of these two forms of materials could potentially maximize their benefits. In this study, porous poly (l-lactide-co-ε-caprolactone) (PLCL) scaffolds were integrated with poly (β-hydroxybutyrate-co-β-hydroxyvalerate) (PHBV) microspheres to enhance the mechanical properties of the scaffolds as well as to potentially regulate cell behavior through altering surface topography. PHBV microspheres fabricated with an emulsion solvent evaporation method were incorporated into PLCL scaffolds (0%, 20%, 40% and 50% W/W). Compressive modulus, surface topography and porosity of the composite scaffolds were evaluated, and in vitro and in vivo chondrogenesis within the chondrocyte-laden scaffolds was investigated by examining proliferation of chondrocytes and the deposition of glycosaminoglycan (GAG) and type II collagen. The results showed significant enhancement of the compressive modulus of the scaffolds incorporated with PHBV microspheres, while Young's modulus of the scaffolds with 50% PHBV incorporation was 3.3 times higher than PLCL scaffolds alone. The porosity of the composite scaffolds was kept constant for all levels of PHBV incorporation. Though the PLCL scaffolds incorporated with microspheres showed no significant effects on adhesion at 6 h as well as in vitro cartilage formation and proliferation of the chondrocytes at both 2 weeks and 4 weeks, total contents of GAG and type II collagen excreted increased significantly with time. The chondrocyte-laden scaffolds formed cartilage-like tissues at 4 and 8 weeks after implantation in nude mice, with increased staining density of type II collagen and GAG over time. In conclusion, incorporation of PHBV microspheres not only enhanced the compressive modulus of PLCL scaffolds, but could also serve as scaffolding structures for cartilaginous tissue formation.