TITANIUM PARTICLES MODULATE MMPs-TIMPs mRNA PRODUCTION IN MG-63 CELL : QUANTITATIVE ANALYSIS BY REAL TIME PCR

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Introduction: Osteolysis is the main cause of joint prosthesis failure. Experimental and clinical works indicate that debris released from the implants is one of the most important factors of osteolysis. Extracellular matrix degradation and connective tissue remodelling around implants have been considered as major biological event in the loosening. The present study was designed to determine the mRNA expression of MMP-1,-2,-9 and TIMP-1,-2 in human osteoblast-like cell with titanium (Ti) particles to clarify one important step in the biological cascade of loosening. Materials and Methods: Human osteoblast-like cells, MG-63, were grown in Dulbecco's Modified Eagle's Medium supplemented with 10% fetal bovine serum, L-glutamate, penicillin, streptomycin and fungizone, at 37 and 5% CO_2 in a humidified atmosphere. Commercially pure different concentration of 0.15% and 0.5% of Ti particles were prepared. Cells were plated at a concentration 4x10^5 incubated for 4 hours. The medium samples were harvested at 6, 12 and 24 hours. Total RNA was extracted by AGPC method and the concentration was adjusted to 1 μg/μl. Enzymatic amplification of the specific cDNA sequences and quantitative analysis were performed using LightCycler system. Results: mRNA expression level of MMP-1,-2 and TIMP-1 ,-2 was suppressed in the presence of 0.5% Ti particles after 6 hours while increased level of mRNA MMP-1,-2 in the presence of 0.5% Ti particle after 24 hours. Expression of MMP-9 was not detected. Discussion: It has been reported that Ti particles can increase the level of cytokines and growth factors relating to bone resorption. Taken together, suppressed mRNA levels of MMPs-TIMPs in the earlier, then increased level of MMPs later indicate that Ti particles can interfere osteoblast function and modulate MMPs-TIMPs production. It may contribute to pathologic matrix degradation in periprosthetic connective tissues, thus leading to osteolysis and implant loosening.
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TITANIUM PARTICLES MODULATE MMPs-TIMPs mRNA PRODUCTION IN MG-63 CELL : QUANTITATIVE ANALYSIS BY REAL TIME PCR

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