Effects of Ti, Al and V Concentrations on Cell Viability

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  • Effects of Ti Al and V Concentrations o

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The effects of Ti, Al and V concentrations and the extraction conditions on the cell viabilities of murine fibroblast L929 cells, murine osteoblast-like MC3T3-E1 cells and fibroblast V79 cells taken from the lungs of Chinese hamsters were investigated by the cell culture method using metallic particles. The particles were sterilized under a U.V. lamp for 14.4 ks (4 h) and suspended independently in Eagle’s minimum essential medium (Eagle’s medium) for the L929 cell seeding and α modified Eagle’s medium (α medium) for the MC3T3-E1 cell seeding containing 10 vol% fetal bovine serum and 7.5% NaHCO3. These mediums were kept for 172.8 ks (48 h) in an incubator under a 95%air-5%CO2 atmosphere (defined as medium extraction). Medium without metallic particles was also kept inside the incubator under similar conditions and was used as the control for all the cell cultures. These mediums were filtered with a 0.2 μm filter. 1.0×104 to 5.0×104 L929 cells/mL, 5×104 MC3T3-E1 cells/mL and 102 V79 cells/dish were seeded in both the control and in the medium extract using the metallic particles. For the titanium particle extraction, the maximum Ti concentrations in the Eagle’s medium and α medium were 0.2 and 0.3 mass ppm, respectively. Hence the L929 and MC3T3-E1 cells proliferated in the same ratio as those of the control. Also, the colony formation ratio (number of colonies in the medium extract with metallic particles/average number of colonies in the control) of V79 cells was unity. For the Al particle extraction with ultrapure water for 172.8 ks (48 h), gel formation of aluminum hydroxide was observed. However, the relative growth ratio (the number of cells in medium extract with metallic particles/average number of cells in control) of the L929 cells was unity when the Eagle’s medium was prepared after filtering with a 0.2 μm membrane filter. On the contrary, the relative growth ratios of the L929 cells for both Eagle’s MEM extract without fetal bovine serum and the Eagle’s medium extract were zero. For the aluminum particle extraction, the relative growth ratios of the L929 and MC3T3-E1 cells sharply decreased as the Al concentration in the medium increased to approximately 0.2 to 0.5 mass ppm. This aluminum effect on L929 cell viability tended to be stronger in the Eagle’s medium containing a small quantity of V ions (≤0.03 ppm). The effects of Al concentration on the colony formation ratio of the V79 cells showed the same tendency as those of the relative growth ratios of the L929 and MC3T3-E1 cells. The relative growth ratio of the L929 cells sharply decreased when V concentration in the Eagle’s medium increased to 0.2 mass ppm and became zero at approximately 0.5 mass ppm. The effects of V concentration on the colony formation ratio of the V79 cells were also similar to the relative growth ratio of the L929 cells. The relative growth ratio of the MC3T3-E1 cells decreased from 0.002 mass ppm of V concentration and approached zero at approximately 0.2 mass ppm.

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