应用等离子体技术聚合制备-NH_2生物膜对大鼠卵巢细胞亲和性的影响

Effect of plasma polymerization prepared -NH_2 biomaterial films on ovarian cell affinity of rats

目的:以丙稀胺为单体,在RF脉冲等离子体条件下聚合制备含氨基的功能薄膜-NH2生物膜,观察应用等离子体技术进行材料表面改性后对卵巢细胞的细胞黏附、增殖及功能状态的影响。方法:实验于2006-11/2007-04在首都医科大学生殖医学研究中心完成。①以丙稀胺为反应单体,利用等离子体处理仪器在盖玻片上聚合薄膜,制备-NH2生物膜基片。②覆有-NH2生物膜的基片为实验组;无覆膜的原始盖玻片为空白基片对照组。将P2代Wistar大鼠卵巢细胞分别接种于两组基片上,在37℃、体积分数为0.05的CO2培养箱中培养。③分别于培养12h和24h两个时间点取样检测。测量基片上细胞黏附面积,计算出百分率,反映细胞黏附率;利用MTT微量酶反应比色法测定MTT代谢率,反映细胞活性;采用放射免疫法检测培养液中雌二醇质量浓度,反映卵巢细胞分泌功能。结果:①-NH2生物膜基片上卵巢细胞的黏附率12h和24h分别为61%,100%,明显高于空白基片上卵巢细胞细胞黏附率47%,78%。②-NH2生物膜基片上卵巢细胞MTT代谢率明显高于对照组(P<0.01)。③-NH2生物膜基片培养液中雌二醇质量浓度也明显高于空白基片(P<0.01)。结论:应用等离子体技术进行表面改性后的-NH2生物膜在较短的时间内对体外培养的大鼠卵巢细胞的黏附、体外存活和分泌功能有明显的促进作用,对大鼠卵巢细胞具有细胞亲和性。

AIM： Using allylamine monomer, prepared amine-containing functional films by radio frequency （RF） pulse plasma polymerization （-NH2 biomaterial films）. This experiment was aimed to study on the effect of -NH2 biomatedal films on ovarian cell adhesion, proliferation and function after material surface modification by plasma technique. METHODS： The experiment was conducted in the Center of Reproductive Medicine, Capital Medical University from November 2006 to Apdl 2007. ①The glasses with -NH2 biomaterial films were prepared by plasma polymerization using allylamine monomer with plasma disposal instrument. ②The test group was composed of the glasses with -NH2 biomaterial films, whereas the control group consisted of the glasses without any treatment. And then Wistar rat ovarian cells of P2 generation were cultured on glasses of each group, at 37 ℃ with the CO2 of 0.05 volume fraction. ③The cell affinity characteristics such as cell adhesion （shown by the proportion of adhered cell） and cell viability （tested by MTT metabolic rate） were evaluated and contrasted at hours 12 and 24 respectively. Culture Medium estrogen （E2） quality concentrations of each group were tested by radioimmunoassay, and it was acceptable to evaluate the secretive function of ovarian cells. RESULTS： ①Adhesion rates of ovarian cells on -NH2 biomaterial films were respectively 61% and 100% at hours 12 and 24, which was significantly higher than those in blank films （47% and 78%）. ②The MTT metabolic rate of ovarian cells on -NH2 biomaterial films was obviously higher than that in the control group （P 〈 0.01）. ③The estrogen （E2） quality concentration in -NH2 biomaterial film culture fluid was markedly higher than that in the control group （P 〈 0.01）. CONCLUSION： The -NH2 biomaterial films have stimulative function on ovarian cell affinity, adhesion, in vitro growth and secretion in a Short time after material surface modification by plasma technique.