Effects of hyaluronic acid and chitosan microspheres on mRNA expression of matrix metalloproteinases-1,-3,-13 and tissue inhibitor of metalloproteinases-1 in the inflammatory chondrocytes

Abstract

Objective To investigate the effect of hyaluronic acid (HA)/chitosan (CS) microspheres on the expression of matrix metalloproteinases (MMPs)-1,-3 and-13 and tissue inhibitor of MMPs (TIMP-1) mRNA in chondrocytes exposed to interleukin (IL)-1 β in vitro,and study the mechanism,then explore the optimal ratio of HA to CS for protecting chondrocytes.Methods HA microspheres,CS nanopaticles and different ratio of HA/CS microspheres were added to culture medium for 24 h,respectively,after 10 μg/L IL-1 β was added to the culture medium of rat chondrocytes and co-cultured for 24 h.The expression levels of MMP-1,-3 and-13,and TIMP-1 mRNA were detected by using reverse transcriptasepolymerase chain reaction (RT-PCR).The concentration of nitric oxide (NO) in the supernatants was measured by Griess reaction.Results HA/CS microspheres could significantly down-regulate the expression of MMP-1,-3,and-13 mRNA (0.467 ±0.072,0.621 ±0.125 and 0.53 ±0.12,respectively) of chondrocytes,while had little effects on the expression of TIMP-1 mRNA (0.69 ± 0.13).Conclusion HA/CS could effectively interrupt the inflammatory cascade reaction caused by IL-1 β through inhibiting NO production. Key words: Hyaluronic acid; Chitosan; Interleukin-1β; Matrix metalloproteinases; Chondrocytes