The induction of MMP-9 release from granulocytes by Vitamin E in UHMWPE

Ultra-high molecular weight polyethylene (UHMWPE) is a biopolymer widely used in orthopaedic implants and its oxidation is considered as major responsible for inflammation and the prosthesis failure. We have studied the effect on the activation of resting human granulocytes of the addition of Vitamin E (Vit.E, α-tocopherol), a natural biological antioxidant and antinflammatory agent, to UHMWPE. We have measured changes in granulocytes morphology and respiratory burst by flow cytometry using Dihydrorhodamine 123 and matrix metalloproteinase 9 (MMP-9, gelatinase B) release and activity in the growth medium using substrate zymography following contact (60min at 37°C) with cell grade polystyrene (PS), normal UHMWPE (PE) and Vit.E added UHMWPE (PE Vit.E). FTIR analyses showed that the surfaces of PE and PE-Vit.E were not significantly different. PS, PE and PE Vit.E did not alter granulocytes morphology and respiratory burst as showed by the mean fluorescence emitted (PS=12.0±0.1, PE=13.0±0.4, PE Vit.E=14.5±0.1). PE Vit.E was able to increase MMP-9 release compared to PS and normal PE (215±16% of the control, p<0.001). The PE Vit.E-induced MMP-9 release was abolished by okadaic acid (0.5nM), suggesting a direct role of Vit.E in the phenomenon.