蚕丝在软骨细胞立体培养中的应用

APPLICATION OF SILKS AS SCAFFOLDS FOR THREE-DEMENSIONAL CULTURE OF CHONDROCYTES

目的 观察蚕丝对软骨细胞的吸附作用及蚕丝对软骨细胞形态和功能的影响。方法 从家蚕蚕茧中抽丝所得的蚕丝经胰蛋白酶消化和聚乳酸包埋 ,制成三维支架 ,软骨细胞与蚕丝三维支架进行复合培养 ,利用相差倒置显微镜、扫描电镜观察软骨细胞生长情况。结果 蚕丝三维支架上滴加软骨细胞悬液后 ,软骨细胞在不规则轻微漂动和缓慢下沉过程中粘附在蚕丝上 ,1～ 2天后完全贴壁。培养 3天后软骨细胞开始分裂 ;5天后 ,细胞生长增殖十分活跃 ;2周时 ,蚕丝三维支架网眼充满软骨细胞。扫描电镜见生长在蚕丝支架上的软骨细胞分泌大量蜘蛛网样基质。结论 蚕丝对软骨细胞具有良好的吸附作用 ,并能维持软骨细胞正常形态和功能 ,蚕丝是软骨细胞立体培养时良好的天然支架。

Objective To observe the effects of silks on attachment, shape and function of chondrocytes cultured in vitro . Methods The silks from silk worm cocoons were digested by trypsin and coated with polylactic acid to from three demensional scaffolds for rabbit rib chondrocyte culture. The growth and shape of chondrocytes were observed with phase contrast microscopy, scanning electron microscopy. Results The chondrocytes were adhered to silks slowly after chondrocytes were seeded into silk scaffolds and cells fixed on silks well 1 or 2 days later. Cells began to proliferate after 3 days and multiplicative growth was observed on the 6th day. Microholes of silk scaffolds were filled with chondrocytes 2 weeks later. Scanning electron microscopy showed that there was a lot of extracellular matrix surrounding cells. Conclusion Silks are ideal for attachment, growth and function maintenance of chondrocytes, and silks can be used as scaffolds for chondrocytes in three demensional culture.