Renò Filippo, Traina Vincenzina, Cannas Mario
Department of Medical Science, Human Anatomy Laboratory, University of Eastern Piedmont, via Solaroli 17, I-28100 Novara, Italy.
J Biomater Sci Polym Ed. 2008;19(1):19-29. doi: 10.1163/156856208783227631.
Matrix metalloproteinases (MMPs) are zinc-dependent enzymes involved in the remodelling of connective tissues during the development and wound healing. Moreover, two MMPs, Gelatinase A (MMP-2) and Gelatinase B (MMP-9), are also present in body fluids such as blood and urine and, therefore, they can be in contact with implanted biomaterials and can be adsorbed onto their surface. In order to test this hypothesis disks of different polymers (polystyrene (PS), polyvinyl chloride (PVC), poly(D,L-lactide) (PLA), polymethyl methacrylate (PMMA) and poly(2-hydroxyethyl methacrylate) (PHEMA)) have been exposed to human plasma and adsorbed proteins have been eluted and analyzed. Using Western blot and substrate zymography analysis, we observed that both MMP-2 and MMP-9 adsorbed onto the surfaces of all the polymers, especially hydrophilic ones (PMMA and PHEMA) and PLA, in both the active and inactive forms. Furthermore, we observed that adhesion of human granulocyte neutophils to PMMA, the polymer that adsorbed the higher quantity of MMP-2 and MMP-9 compared to the others, was reduced by more that 50% by the presence of a gelatinase inhibitor. This data suggest a surprising role of these absorbed enzymes in the adhesion of neutrophil onto some polymeric biomaterials surface and, therefore, in the setting of inflammation.
基质金属蛋白酶(MMPs)是锌依赖性酶,参与发育和伤口愈合过程中结缔组织的重塑。此外,两种基质金属蛋白酶,明胶酶A(MMP-2)和明胶酶B(MMP-9)也存在于血液和尿液等体液中,因此,它们可与植入的生物材料接触并吸附在其表面。为了验证这一假设,将不同聚合物(聚苯乙烯(PS)、聚氯乙烯(PVC)、聚(D,L-丙交酯)(PLA)、聚甲基丙烯酸甲酯(PMMA)和聚(甲基丙烯酸2-羟乙酯)(PHEMA))制成的圆盘暴露于人体血浆中,然后洗脱并分析吸附的蛋白质。通过蛋白质印迹法和底物酶谱分析,我们观察到MMP-2和MMP-9均以活性和非活性形式吸附在所有聚合物表面,尤其是亲水性聚合物(PMMA和PHEMA)以及PLA表面。此外,我们观察到,与其他聚合物相比,PMMA吸附的MMP-2和MMP-9量更高,而明胶酶抑制剂的存在使人类粒细胞中性粒细胞与PMMA的黏附减少了50%以上。这些数据表明,这些吸附的酶在中性粒细胞黏附于某些聚合物生物材料表面以及炎症发生过程中发挥了惊人的作用。