Effect Of Cu/LDPE Nanocomposite Material On Human Endometrial Cells In Vitro And Its Clinical Trial

Part I Effect of Cu/LDPE Nanocomposite Material Leaching Liquor on Proliferative Activity, Cell Cycle and Apoptopsis of Human Endometrial Cells in vitroTo explore the effect of Cu/LDPE nanocomposite material leaching liquor (Nano Cu )on human endometrial cells in vitro, proliferative activity, cell Cycle and apoptopsis of human endometrial cells were observed. This part included three experiments:Experiment one Purification and Culture of Human Endometrial Cells in vitroObjective: To establish a method of human endometrial cells culture,storage and subculture in vitro.Methods: Endometrial glandular cells and stromal cells were harvested by collagenase digestion and copper sieve filtration methods, and monolayer-cultured with RPMI1640.Results: Glanduar cells and stromal cells in the culture had been observed by light microscopy and stained positively by keratin and vimentin McAb respectively using immunohistochemical method.The life span of glandular cells lasted 4-6 weeks and 2-3 passages;the life span of stromal cells lasted 5-8 weeks and 4-6 passages.Conclusions: Method of human endometrial cells culture,storage and subculture in vitro has been established, it's prepared for the later treament experiment on it.Experiment two Effect of Cu/LDPE Nanocomposite Material Leaching Liquor on Cell Cycle and Apoptopsis of Human Endometrial Cells in vitroObjective: To explore the effect of leaching liquor of Cu/LDPE nanocomposite material (Nano Cu ) on cell cycle and apoptosis of human endometrial cells in vitro.Methods: The human endometrial cells were cultured and subcultured in vitro. The endometrial cells of the second generation were treated with Cu/LDPE nanocomposite material leaching liquor with different Cu2+ concentrations(experiment groups) and RPMI 1640 complete medium(control group). The cell cycle and apoptosis rates were detected by PI staining and flow cytometry.Results: Compared with those in the control group, the cell apoptosis rates were increased in experiment groups, especially in the experiment groups with high concentrations of Cu2+ (1.25μg/ml, 5μg/ml and 20μg/ml) (p<0.05); Compared with those in the control group, the G0/G1 phase cells were increased and the S phase cells were decreased distinctly in experiment groups with high concentrations of Cu2+ (0.3125μg/ml, 1.25μg/ml, 5μg/ml and 20μg/ml) (p<0.05). There was no significant difference between experiment group with Cu2+ concentration of 0.078125μg/ml and control group.Conclusions: The reduplication of human endometrium cells could be blocked in G0/G1 phase,and its apoptosis rates could be obviously enhanced by Nano Cu at high Cu2+ concentration, which may suggested the damage of endometrium cells could be induced by high-concentration Nano Cu. Experiment three Effect of Cu/LDPE Nanocomposite Material Leaching Liquor on Proliferative activity of Human Endometrial Cells in vitroObjective: To explore the effect of leaching liquor of Cu/LDPE nanocomposite material (Nano Cu )on proliferative activity of human endometrial cells in vitro.Methods: The proliferative index of endometrial cells were detected by CFSE cell staining and flow cytometry at 36 hours and 72 hours after treatment of Nano Cu with different Cu2+ concentrations (experiment groups) and RPMI 1640 complete medium(control group) respectively.Results: Compared with those in the control group, proliferative indexes of endometrial cells were decreased obviously in experiment groups with Cu2+ concentrations more than 0.3125μg/ml, with the differences being statisticly significant (p<0.01). Proliferative index decreased distinctly with increase of concentration of Cu2+; but there was no significant difference between experiment group with Cu2+ concentration of 0.078125μg/ml and control group.Conclusions: The proliferative activity of human endometrium cells might be inhibited distinctly by Cu/LDPE nanocomposite material leaching liquor. There might be concentration-response relationship between Cu2+ concentrations and inhibition of human endometrium cells'proliferative activity. Part II Clinical Observation to Acceptability of Copper/Low Density Polyethylene Nanocomposite IUDPurpose: The study was conducted to compare antifertility effectiveness and side effects of Copper/low density polyethylene nanocomposite IUD (experimental group) and Copper T220C IUD (control group).Methods: 100 females were randomly divided into two groups (experimental group and control group, n=50 in each group). Clinical observation and comparative study were performed on two groups for 12 months.Results: Follow-up rate was 100% at 12th month. In experimental group and control group, the cumulative continuation rates were both 92.0 per 100 women at 12th month and there was no difference between them (P>0.05). The pregnancy rate, removal rate and expulsion rate were low with the difference being not statistically significant too (P>0.05). The most common side-effects were large menstrual bleeding, spotting and pain. The rates of side-effects were lower in the experimental group than control group, especially in the initial 3 months after insertion with the differences being statistically significant (P<0.05).Conclusions: The new design of the copper/low density polyethylene nanocomposite IUD showed low pregnancy rate, high contraceptive efficacy and satisfactory acceptabilily. Compared with those of TCu220C IUD, the rates of side-effects were lower in copper/low density polyethylene nanocomposite IUD group.