Objective It is to explore the effect of Bufalin/chitosanano-hydroxyapatite ( bufalin/CSano - HA ) scaffolds on proliferation of human osteosarcoma cells in vitro, hope to find novel approaches of chemotherapy and reconstruction for osteosarcoma. Methods Bufalin/CSano - HA scaffolds were prepared by freezing and lyophylization of composite solution. Simulated body fluid ( SBF ) soaking test in vitro was used to investigate the drug release. Confocal laser scanning microscopy ( CLSM ) was used to observe 3 - dimensional configuration inside the scaffold by acridine orange/ethedium bromide ( AO/EB ) fluorescence method. MTT assay was used to evaluate the proliferation of osteosarcoma cells on the scaffolds. Results The effective releasing time of bufalin/CSano - HA was 16 days. Majority of the cells on the scaffolds were randomly attached on the material surface while the others scattered as masses. They exhibited the spheric growth with karyopyknosis and green colour. The inhibitory rate of cells in the composite scaffolds with 100 mg/L bufalin was significantly higher than those with 50 mg/L bufalin groups and 25 mg/L bufalin groups from the 4th day in culture ( P ＜ 0.05 ). Conclusion Bufalin/CSano - HA scaffolds can significantly inhibit the proliferation of osteosarcoma cells in vitro. It could provide a new idea in the clinical therapy for osteosarcoma.%目的 探讨复合蟾毒灵的壳聚糖/纳米羟基磷灰石人工骨对骨肉瘤细胞增殖的影响,希望为骨肉瘤的化疗和修复重建找到新的思路.方法 冷冻干燥法制备复合人工骨,通过体外模拟液浸泡实验观察蟾毒灵在体外缓释情况;激光共聚焦显微镜观察细胞三维生长分布的状态;MTT法检测复合人工骨对骨肉瘤细胞增殖的影响.结果 蟾毒灵有效释药时间为16d;细胞在支架上多为游离分布,球形,核固缩;并且从培养的第4天开始含100mg/L蟾毒灵组细胞抑制率＞含50mg/L蟾毒灵组＞含25mg/L蟾毒灵组.结论 复合蟾毒灵的壳聚糖/纳米羟基磷灰石人工骨能够长期持续抑制骨肉瘤细胞的增殖,可能为临床治疗骨肉瘤提供一种新的思路.
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