软组织填充材料聚氨酯微球的制备及体外生物相容性研究  

PREPARATION AND BIOCOMPATIBILITY OF POLYURETHANE MICROSPHERES FOR BIOMEDICAL APPLICATIONS

作　　者：笪琳萃[1] 王旻[1] 龚梅[1] 张利[2] 解慧琪[1] 

机构地区：[1]四川大学华西医院再生医学研究中心.干细胞与组织工程研究室,成都610041 [2]四川大学分析测试中心/纳米生物材料研究中心

出　　处：《中国修复重建外科杂志》2015年第5期626-631,共6页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金资助项目(51443003);中国博士后科学基金资助项目(2013M531971)~~

摘　　要：目的制备生物医用聚氨酯微球,对其理化性能及体外生物相容性进行评价。方法采用自乳化法(乳化速率分别为1 000、2 000、3 000、4 000 r/min)制备聚氨酯微球,采用傅里叶变换红外光谱仪测试聚氨酯微球的分子结构,扫描电镜观察微球内部结构及表面形貌,筛选最佳乳化速率制备的微球进行后续实验。培养人脐静脉内皮细胞(human umbilical vein endoth elial cells,HUVECs),采用钙黄绿素/碘化丙啶(calceinacetoxymethylester/pyridine iodide,Calcein-AM/PI)双染观察细胞在聚氨酯材料上的形态及生长黏附情况。分别用含1%苯酚、10%FBS的H-DMEM培养基(A组)、按GB/T 16886.12标准规定比例制备的聚氨酯材料浸提液(B组)、含10%FBS的H-DMEM培养基(C组)培养细胞,采用细胞计数试剂盒8(cell counting kit 8,CCK-8)法评价细胞活力;通过溶血实验评价血液相容性,其中浸提液作为实验组,生理盐水作为阴性对照组,蒸馏水作为阳性对照组。结果扫描电镜观察示,乳化速率为2 000 r/min时制得的微球形态大小均较为理想,聚氨酯涂膜的表面为粗糙致密结构,内部为不均匀的多孔结构。Calcein-AM/PI双染观察示HUVECs与聚氨酯材料黏附紧密,且以绿染的活细胞为主。CCK-8检测示B、C组各时间点细胞增殖活力均显著高于A组(P<0.05),材料细胞毒性低。溶血实验测定示,阳性对照组吸光度(A)值为0.864±0.002,阴性对照组A值为0.015±0.001,聚氨酯微球浸提液的溶血率为0.39%±0.07%(<5%标准),无溶血作用。结论通过自乳化法成功制备了球形度较好的聚氨酯微球,该材料有利于细胞黏附增殖、细胞毒性低,且具备良好血液相容性,有望用作软组织缺损填充材料。Objective To prepare polyurethane（PU） microspheres and evaluate its physicochemical properties and biocompatibility for biomedical applications in vitro.Methods The PU microspheres were prepared by selfemulsification procedure at the emulsification rates of 1 000,2 000,3 000,and 4 000 r/min.The molecular structure was tested by Fourier transform infrared spectrometer and the surface and interior morphology of PU microspheres were observed by scanning electron microscopy（SEM）.PU microspheres prepared at best emulsification rate were selected for the subsequent experiment.The human umbilical vein endothelial cells（HUVECs） were cultured and seeded on the materials,then cell morphology and adhesion status were observed by calcein-acetoxymethylester/pyridine iodide（Calcein-AM/PI） staining.The cells were cultured in the H-DMEM containing 10%FBS with additional 1% phenol（group A）,in the extracts of PU prepared according to GB/T 16886.12 standard（group B）,and in H-DMEM containing 10%FBS（group C）,respectively.Cell counting kit 8（CCK-8） assay was used to detect the cell viability.The blood compatibility experiments were used to evaluate the blood compatibility,the PU extracts as experimental group,stroke-physiological saline solution as negative control group,and distilled water as positive control group.The hemolytic rate was calculated.Results The SEM results of PU microspheres at the emulsification rate of 2 000 r/min showed better morphology and size.The microstructure of the PU was rough on the surface and porous inside.The Calcein-AM/PI staining showed that the HUVECs attached to the PU tightly and nearly all cells were stained by green.CCK-8 assays demonstrated that group B and group C presented a significantly higher cell proliferative activity than group A（P〈0.05）,indicating low cytotoxicity of the PU.The absorbance value was 0.864±0.002 in positive control group and was 0.015±0.001 in negative control group.The hemolysis rate of the PU extracts was 0.39%±0.07%（〈5%）,i

关 键 词：聚氨酯微球 人脐静脉内皮细胞 软组织填充材料 血液相容性 

分 类 号：R318.08[医药卫生—生物医学工程]