Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) P(3HB-co-4HB) biopolymer was subjected to a depyrogenation treatment followed by blending with vitamin E (P(3HB-co-4HB)/Vit E blend), collagen (Col blend) or both (P(3HB-co-4HB)/Vit E/Col hybrid blend) to improve its biological performance. The cytocompatibility and pro-inflammatory response to the biopolymer after the treatment and blending were evaluated by cellular responses in vitro. Cell attachment and spreading of murine NIH 3T3 fibroblasts seeded on the films were found to be the highest on the hybrid P(3HB-co-4HB)/Vit E/Col blend film. The P(3HB-co-4HB) blends showed increased cell proliferation and negligible cytotoxicity. The pro-inflammatory cytokines, IL6 and TNF α expression in the murine macrophage cell line J774 A-1 monocyte/macrophage cells cultured on the biopolymer films were assessed by immunoblotting and RT-PCR analyses which revealed that the combined blending strategy was the most effective in reducing the pro-inflammatory activity. The depyrogenated P(3HB-co-4HB) blends significantly extend the range of biomaterials suitable for tissue engineering.
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