摘要:
将猪松质骨脱脂脱蛋白、煅烧后再进行球磨制备羟基磷灰石(HA), 与壳聚糖(CS)共混制备成HA/CS复合膜。通过正交设计考察了球磨条件对HA粒径的影响, 在复合膜上接种MC3T3-E1前体成骨细胞, 采用SEM及四甲基偶氮唑盐(MTT)方法检测细胞在膜表面的形态及增殖, 评价了材料的细胞相容性。结果表明, 所制备HA为粒径较均一的微米级球形粒子, 中值粒径D50为1.21~1.67 μm。共混膜内HA在基质中分布均匀, 二者结合紧密, 复合膜具有较好的力学性能, MC3T3-E1细胞能在复合膜上很好地黏附生长。细胞增殖结果表明, 复合膜制备条件煅烧温度为1000 ℃, 球配比4∶4∶2, 球磨速率为230 r·min-1, 球磨时间为2.5 h, HA∶CS =5∶5 (质量比)时最利于细胞增殖。
Abstract:
Hdroxyapatite(HA) was prepared using porcine trabecular bone after being degreased, deproteined, calcined and ball milling. HA/chitosan(CS) composite membranes were prepared by blending and drying HA and CS solution. The effect of milling conditions on the HA particle size was inspected by the orthogonal design. MC3T3-E1 preosteoblasts were seeded on the surface of the composite membranes, and then cell morphology and proliferation were detected by SEM and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide(MTT) methods. The results show that the prepared HA is relatively homogeneous micrometer-scale spherical particle with median diameter(D50) in 1.21~1.67 μm range, and HA particles are distributed uniformly in the matrix, HA particle and CS combined closely. Composite membrane has good mechanical properties, MC3T3-E1 cells can adhere and grow well on the surface of composite membrane. The cell proliferation results show that composite membrane under the preparation conditions: calcination temperature 1000 ℃, the milling ball propotion 4∶4∶2, milling rate 230 r·min-1, milling time 2.5 h, HA∶CS =5∶5 (mass ratio) is the most beneficial for cell proliferation.
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