目的：通过体外细胞培养试验，对新型的纳米羟基磷灰石／聚乙烯醇（HA／PVA）多孔复合水凝胶人工角膜支架材料的细胞毒性和生物相容性进行研究。方法将试验材料分为4组：大孔径的纳米HA／PVA复合水凝胶、小孔径的纳米HA／PVA复合水凝胶、大孔径的PVA水凝胶及小孔径的PVA水凝胶材料，所有大孔径为200μm ，小孔径为100μm。分别用4组材料的浸渍液培养兔角膜成纤维细胞，空白对照组不加入材料直接在培养基中培养。通过MTT 比色法检测各组吸光值（A），并计算各组细胞相对增值率（RGR），对4组材料的细胞毒性进行评价。同时将兔角膜成纤维细胞在4组材料表面直接培养，进行荧光蛋白转染后，72 h于倒置显微镜下观察各组成纤维细胞在培养板上增殖生长情况，并通过扫描电镜观察成纤维细胞在材料上黏附生长，以及向孔隙内迁移的情况。结果大孔径的纳米 HA／PVA复合水凝胶材料和大孔径的PVA水凝胶材料毒性评分均为1级无细胞毒性。72 h倒置显微镜下培养板每视野细胞计数，大孔径组与空白对照组差异无统计学意义（ P＝0．608）。72 h扫描电镜下观察，大孔径及小孔径纳米 HA／PVA复合材料表面见兔角膜成纤维细胞黏附、生长增殖，同时向材料孔隙内迁徙；每视野细胞计数，组间差异有统计学意义（F＝95．245，P＝0．000），大孔径纳米 HA／PVA 复合材料组细胞黏附显著多于其他组。结论纳米HA／PVA 多孔复合水凝胶材料无明显细胞毒性，同时具有良好的生物相容性，是一种具有良好临床应用前景的人工角膜支架材料。%Objective To evaluate the biocompatibility of nano-hydroxyapatite/polyvinyl alcohol (n-HA/PVA ) hydrogel com-posite for keratoprosthesis .Methods There were four materials to be evaluated ,big-porous and small-porous n-HA/PVA hydrogel composite ,big-porous and small-porous PVA hydrogel .Every kind of materials was digested in F12 nutrient culture for 24 h to get the materials impregnant .Corneal fibroblasts were cultivated in materials impregnant and through methyl thiazolyl tetrazolium col-orimetry to assay the cytotoxicity .Rabbit corneal fibroblasts were seeded onto the four materials in vitro .Three days later the growth of fibroblasts were observed by inverted microscope and scanning electron microscope (SEM ) .Results The cytotoxicity scores of the materials with big-porous were both grade one .The mean corneal fibroblasts counts on culture were no obvious differ-ence between big-porous materials groups and control group under plate inverted microscope 72 hours later(P=0 .608) .Through scanning electron microscope 72 hours later ,corneal fibroblasts grew well on n-HA/PVA hydrogel composite ,and migrated into the porous of composite materials .The corneal fibroblasts on big-porous n-HA/PVA hydrogel composite were more obviously found than the other three materials(F=95 .245 ,P=0 .000) .Conclusion The porous n-HA/PVA hydrogel composite is a kind of good biocompatible material which has no cytotoxicity .The n-HA/PVA hydrogel composite with big porous would be an ideal pe-ripheral skirt of keratoprosthesis .
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