纳米羟基磷灰石人工心脏机械瓣膜细胞相容性的研究

目的探讨利用脉冲激光沉积（PLD）方法在人工心脏机械瓣膜上沉积的纳米相羟基磷灰石（HA）薄膜的组织相容性，为人工心脏瓣膜的构建提供理想的支架薄膜材料。方法将传代的人血管内皮细胞制成细胞悬液接种在HA材料上，置含5％（V／V）CO2、37℃的恒温培养箱内静态培养3周。分别在培养3、7、14和21d后取出部分材料，用2．5％戊二醛固定，乙醇梯度脱水，临界点干燥、喷金，于扫描电子显微镜（SEM）下观察细胞在材料上的附着情况。用四氮唑盐比色（MTT）法测定血管内皮细胞与材料复合培养的增殖能力。结果扫描电镜观察复合培养，见HA材料的表面有细胞附着且数目逐渐增多，到第21天时可见细胞融合成片，并将材料表面覆盖，部分区域有细胞外基质形成。MTT法评价材料对血管内皮细胞的细胞毒性，见细胞增殖未受到明显影响，生长良好。结论HA对血管内皮细胞的增殖和分化功能无明显影响，对细胞无明显毒性，具有良好的生物相容性，可以作为心脏瓣膜材料安全应用。

The biocompatibility research study on mechanical heart valve of hydroxyapatite ( HA ) film complex materials and human blood vessel endothelial cells

Objective We make use of impulse laser sediment （PLD） method to make nanometeric hydroxyapatite （HA）film sedimenting on the surface of artificial heart valve, exploring the biocompatibility of nanometeric HA film complex materials to investigate biocompatibility of the hydroxyapatite, we are searching ideal film materials for mechanical heart valve applications. Methods Transferred Human vascular endothelial cells suspension was implanted on hydroxyapatite materials, and was placed in the incubator contained 5% （V/V） CO2 for 3 weeks. A few materials were took out at 3, 7, 21 days respectively and were fixed with 2.5% glutaraldehyde. The samples were dried with increasing concentrations of ethanol successively. The materials were characterized using scanning electron microscopy （SEM） to observe cell on the materials and determined the cell growth conditions with MTT chromatometry. Results SEMs of Human vascular endothelial cells seeded onto the HA surface showed good cell morphology and coverage of the HA surface. Conclusion The hydroxyaptite has an excellent biocompatibitity and safely for for mechanical heart valve applications.