摘要
背景：清华大学材料科学与工程系冯庆玲教授等采用仿生学原理,制成一种塑形简便并且具有良好骨传导、骨诱导性的纳米羟基磷灰石/壳聚糖复合骨修复材料。但制备的复合材料是否仍具良好的生物相容性尚不确切。目的：观察纳米羟基磷灰石/壳聚糖复合材料与兔骨髓间充质干细胞的相容性。设计、时间及地点：体外观察实验,于2008-08/11在南方医科大学珠江医院中心实验室完成。材料：纳米羟基磷灰石/壳聚糖复合材料由清华大学材料系研制,孔隙率90%,孔径50~200μm。骨髓间充质干细胞由2周龄健康新西兰大白兔股骨和胫骨骨髓经密度梯度离心法获得。方法：将预湿后的纳米羟基磷灰石/壳聚糖材料置于培养板内,将第3代骨髓间充质干细胞接种于材料表面在体外复合培养。对照组为单纯骨髓间充质干细胞培养。主要观察指标：倒置显微镜、扫描电镜观察细胞和材料复合培养后细胞生长状况,计数细胞接种后1,2,4h在材料表面的黏附状况,CCK-8检测细胞在复合材料上的增殖状况,流式细胞仪检测种植细胞的细胞周期。结果：骨髓间充质干细胞在复合材料表面上生长状况良好。细胞接种到复合材料1h时黏附率低于对照组（P〈0.05）,但接种2,4h后两组黏附率差异无显著性意义（P〉0.05）。细胞接种后,两组均保持正常的分裂增殖速度（P〉0.05）。材料组和对照组细胞皆为正常的二倍体细胞,未见异倍体细胞形成,复合材料对兔骨髓间充质细胞的细胞周期影响不大。结论：纳米羟基磷灰石/壳聚糖与骨髓间充质干细胞具有良好的生物相容性。
BACKGROUND： The composite of nano-hydroxyapatite/chitosan （nHA/CS）, which prepared by Professor Feng Qin-ling, has good biocompatibility and osteoconductive ability. However, the biocompatibility of the composite is poorly understood. OBJECTIVE： To evaluate the biocompatibility between nHA/CS composite and rabbit bone marrow stem cell （BMECs） in vitro. DESIGN, TIME AND SETTING： An in vitro observation was performed at the Central Laboratory of Zhujiang Hospital, Southern Medical University from August to November2008. MATERIALS： The nHA/CS composite with porosity of 90%, pore size 50 200 μ m, was provided by Department of Materials, Qinghua University. BMECs were obtained from 2-week-old healthy rabbit＇s femur and tibia bone marrow by density gradient centrifugation. METHODS： The nHA/CS composite was co-cultured with the third passage of BMECs at a culture plate. The culture BMECs group was served as the control group. MAIN OUTCOME MEASURES： The growth of cells was observed by contrast phase microscope and scanning electron microscope. Cell attachment on nHNCs was count at 1, 2, and 4 hours after seeding. Proliferation of cell was analyzed by CCK-8 assay. In addition, the cell cycle was detected by flow cytometry. RESULTS： The BMSCs grow well on the surface of nHA/CS. The attachment rate in the nHA/CS group was lower than that of the control group at 1 hour （P 〈 0.05）; however, there was no significant difference after 2 and 4 hours （P 〉 0.05）. After seeding, the proliferation of cell had no changes （P 〉 005）. Normal diploid cells could be found in both groups, which indicated that nHA/CS had no influence to cell cycle. CONCLUSION： nHA/CS has an excellent biocompatibility with BMECs in vitro.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第8期1423-1426,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
广东省自然科学基金资助项目(7005193)~~
