摘要
目的制备羟基磷灰石修饰的多壁碳纳米管(HA-MWCNTs),探索HA-MWCNTs对大鼠骨髓间充质干细胞(BMSCs)的毒性作用及其多向分化能力的影响,为HA-MWCNTs在组织工程中的应用提供试验依据。方法制备HA-MWCNTs,通过透视电镜、X线衍射分析其微观形态;取体重100~150 g的4周龄雌性SD大鼠骨髓间充质干细胞,利用体外细胞培养技术,将第3代BMSCs与HA-MWCNTs或未修饰的多壁碳纳米管(MWCNTs)共培养,通过MTT法及活死细胞检测评估碳纳米管对BMSCs的细胞毒性;通过诱导其脂向及骨向分化,评估碳纳米管对BMSCs多向分化能力的影响。结果与对照组比较,Raw-MWCNTs对细胞增殖有明显抑制作用(P<0.05),而HA-MWCNTs对细胞增殖活力影响较小(P>0.05);活死细胞检测提示HA-MWCNTs组死细胞率较低。HAMWCNTs使BMSCs骨向分化相关基因Cbfal/Runx2、BSP、OCN的信使RNA升高(P<0.05),脂向分化相关基因a P2、脂蛋白脂酶的信使RNA降低(P<0.05)。结论与MWCNTs比较,HA-MWCNTs对细胞的增殖影响较小,细胞毒性较低,生物相容性良好。HA-MWCNTs可促进BMSCs骨向分化,抑制其脂向分化。其在骨组织工程领域中的应用具有良好前景。
Objective To develop hydroxyapatite functionalized multi-walled carbon nanotubes（HA-MWCNTs）, and assess the toxicity of HA-MWCNTs on rat＇s BMSCs and its influence on the multipotent differentiation of bone marrow derived mesenchymal stem cells（BMSCs）, thereby to provide experimental evidence for its utility in tissue engineering.Methods HA-MWCNTs were synthesized. Transmission electron microscope（TEM）, X-ray diffraction（XRD） were used to observe the microstructure; BMSCs were obtained from female SD rats of 4 weeks old which weighted between 100 to150 grams. After cultured and expanded to the third passage, cells were co-cultured with HA-MWCNTs and unfunctionalized MWCNTs（MWCNTs）. MTT and live-dead cell assay were used to appraise the toxicity of MWCNTs on BMSCs. BMSCs were then induced and differentiated into adipocytic and osteoblastic cells and were used to assess the influence of MWCNTs on BMSCs＇ multipotent differentiation. Results Compared to control group, BMSCs in MWCNTs group were significantly inhibited（P〈0.05）, while in HA-MWCNTs group, BMSCs were slightly inhibited, with no statistical difference（P〉0.05）, and the ratio of dead cells was less in HA-MWCNTs group. The m RNA level of osteogenesis（Cbfal/Runx2, BSP and OCN） was increased（P〈0.05） and the m RNA of adipogenesis（aP2 and LPL） was decreased（P〈0.05） in HA-MWCNTs group. Conclusion Compared with MWCNTs, HA-MWCNTs had slight influence on cell proliferation, trivial toxicity and favorable biocompatibility of BMSCs, meanwhile it could promote osteoblastic and inhibit adipocytic differentiation of BMSCs. HA-MWCNTs may supply a novel platform for applications of scaffold in bone tissue engineering.
作者
宋国栋
郭小双
宗宪磊
赵竟伊
赖晨智
靳小雷
SONG Guodong;GUO Xiaoshuang;ZONG Xianlei;ZHAO Jingyi;LAI Chenzhi;JIN Xiaolei(Department No.16,Plastic Surgery Hospital,Chinese Academy of Medical Science,Beijing 100144,China)
出处
《中国医药导报》
CAS
2018年第24期4-8,13,共6页
China Medical Herald
基金
国家自然科学基金资助项目(81400565)
关键词
羟基磷灰石
碳纳米管
骨髓间充质干细胞
多向分化
Hydroxyapatite
Carbon nanotube
Bone-marrow derived mesenehymal stem cell
Muhipotent differentiation
