The Research Of CeO₂ Nanoparticles Effect On Alveolar Macrophages NR8383 And Alveolar Epithelial Cells RPAEpiC In Rat

Objective In this experiment,rat alveolar macrophages NR8383 and rat alveolar epithelial cells RPAEpi C were chosen as the research objects to study the biological effects and toxicity organism produced by nano-cerium oxide in the cellular level in vitro.Method Cells were put into 37 ? and 5% CO₂ incubator to culture,then different concentration or size or style cerium oxide nanoparticles were added.Dose-effect experiment: 20 nm Ce O₂ of 5,10,20,50,100mg/ml concentration were added.Size-effect experiment: 100mg/ml 35 nm,68nm,287 nm,1-5mm Ce O₂ were added.Comparation experiment 1: 20 nm Ce O₂,34 nm Zn O,24 nm Si O₂ were added.Comparation experiment 2: 287 nm Ce O₂,300 nm Nd2O3 were added.10 ml serium-free culture medium were added into plates as blank control group?0mg/ml?.After 24 h,to detect the cell survival rate,cell permeability,oxidative stress,inflammatory factors,apoptosis,cell cycle,phagocytosis.Results?1?The research of nano-cerium oxide effect on RPAEpi C cells:?1?Dose-effect study: the survival rate had a dose-effect,it rose with the increasing dose;nano-cerium oxide made LDH content gradually;phagocytosis significantly enhanced,the amount of particles uptaken gradually increased with the increasing dose,when the cells uptaked 50 mg/ml,it reached saturation;apoptosis rate decreased: at the concentration of 5,10,20mg/ml the capacity of inhibitting apoptosis gradually increased,at the concentration of 50,100mg/ml the ability of inhibiting apoptosis was weaker than that at the concentration of 5,10,20mg/ml.At the concentration of 5,10,20mg/ml SOD content gradually increased,at the concentration of 50,100mg/ml SOD content gradually decreased.?2?Size-effect study: the survival rate rose with the increasing dose,but the difference was not significant;the ACP content significantly increased and the phagocytosis significantly increased: phagocytosis reduced with the size of particles increasing;there were particles in cells,the amount of particles that cells obsorbed had no significant difference in nano-size,but the amount of particles that cells obsorbed was less in micron-size than that in nano-size;the LDH contents both increased;the SOD content increased obviously when the sizes were 35 nm and 387 nm,and the SOD content of micron-cerium oxide was lowest,the size-effect was not obvious;the NO contents were both decreased in nano-size and the NO content was increased in micron-size.?3?100mg/ml three kinds of nano-oxide effected on rat alveolar epithelial cells 24 h,nano-cerium oxide made the NO content and the cell viability increase,the membrane permeability increased,cells had no injury.Nono-Zinc Oxide changed cells morphology,the NO content increased,the cell viability dropped,the toxicity was extremely huge.Nano-silicon oxide could also be obsorbed,membrane permeability and the NO contents increased,but it didn't have enough toxicity to cell survival rate.?4?Two kinds of nanometer rare earth oxide effected on RPAEpi C cells,nano-cerium oxide promoted cell proliferation and had less effect on membrane permeability.Neodymium oxide nanoparticles greatly reduced cell viability,LDH leaked serious from cytoplasm,toxic effect was large.?2?The research of nano-cerium oxide effect on NR8383 cells:?1?Dose-effect study: At the concentration of 10-100mg/ml the survival rate rose with the increasing dose,the survival rate reached the highest rate at the concentration of 100mg/ml,nano-cerium oxide didn't produced inhibition to cells in this range of concentration.Compared to the control group,LDH contents both increased,each concentration of nano-cerium oxide enlarged membrane permeability,at the concentration of 10,20,50mg/ml cell permeability increased with the increasing dose,at the concentration of 100mg/ml the change of membrane permeability was the smallest,the dose-effect was not obvious.Compared to the control group,at the concentration of 10,20,50mg/ml the SOD content decreased,at the concentrations of 5mg/ml and 10mg/ml the SOD content increased,suggestting a trend that a higher concentration or lower concentrations injuried weak.At the concentration of 10,20,50mg/ml nano-cerium oxida made NO content decreased slightly,the NO content was lower than the control group,the NO contents were similar to the control group when the concentrations were 5mg/ml and 100mg/ml.When 6h,under the effct of 0-5mg/ml concentration,cerium oxida nanoparticles promoted apoptosis,then the effect of inhibiting apoptosis increased with the increasing dose;When 12 h,at the concentration of 0-10mg/ml cerium oxida nanoparticles promoted apoptosis with the increasing dose,then the effect of inhibiting apoptosis increased with the increasing dose;When 24 h,0-10mg/ml cerium oxida nanoparticles promoted apoptosis with the increasing dose,then the effect of inhibiting apoptosis decreased with the increasing dose,at last inhibited apoptosis and the effect of inhibiting apoptosis increased with the increasing dose.?2?Size-effect study:Nano-size cerium oxide made the survival rate increased,1-5mm cerium oxide inhibited cell proliferation;each size group both increased the LDH content,suggesting that they made membrane permeability enlarged,the change of membrane permeability was the biggest in 1-5mm group.Nano-size cerium oxide group made the NO content decreased,1-5mm cerium oxide group made the NO content decreased.It showed that nano-size cerium oxide group had no significant cytotoxicity and micron ceria group might have cytotoxicity.?3?Study on the comparation of three kinds of nano-oxide: nano-cerium oxide made the NO content decreased and had no toxicity to cells.Nano-silica made the NO content increased and had a weak toxicity.Nano-Zinc Oxide made the NO content increased and had a significant toxicity.Conclusions Nano-cerium oxida effect on rat alveolar epithelial cells RPAEpi C and rat alveolar macrophages NR8383 24 h,dose-effect is not obvious: in selected concentration range,nano-cerium oxida do not show significant cell injury,particles can be both taken into cells,the promoting effects are lighter than other concentrations to cells when the concentrations are 5mg/ml and 100mg/ml.Size-effect is not obvious: nano-size cerium oxide group have no significant toxic effecting on the cells,micron-size cerium oxide group may cause toxic effecting on the cells,particles can be both obsorbed by cells,micron particles are obsorbed less than nanoparticles.Compared to other similar nano-oxide particles,cerium oxide nanoparticles have no significant toxicity on cells,nano-silica have weak toxicity,nano-zinc oxide have significant toxicity.Compared with the similar size of the nano-rare earth oxide,cerium oxide nanoparticles have no significant cytotoxicity,neodymium oxide nanoparticles show significant cytotoxicity.