纳米晶胶原基骨修复材料与兔骨髓间充质干细胞体外复合培养

背景:纳米晶羟基磷灰石胶原基骨修复材料具有与天然骨成分接近,结构和形貌图谱分析与天然骨相似,生物相容性好等特点,且材料多孔,孔径较大、孔隙率及孔隙交通率高,符合作为骨组织工程支架材料的要求.目的:观察兔骨髓间充质干细胞与纳米晶胶原基骨修复材料体外复合培养的结合程度.设计、时间及地点:体外观察实验,于2006-03/09在江苏大学医学院细胞实验室完成.材料:健康雄性新西兰大白兔1只,1月龄.纳米晶胶原基骨修复材料由清华大学材料系提供.方法:体外分离培养、纯化兔骨髓间充质干细胞,取第3代骨髓间充质干细胞与纳米晶胶原基骨体外复合培养.主要观察指标:①兔骨髓间充质干细胞生长形态及生长曲线.②复合培养5,10 d后扫描电镜观察二者复合程度.③复合培养5,10 d时纳米晶胶原基骨修复材料上结合的细胞数量.结果:兔骨髓间充质干细胞贴壁生长,增殖速度快,生长曲线符合Logistic生长曲线,兔骨髓间充质干细胞与纳米晶胶原基骨体外复合培养,在第5,10天进行扫描电镜观察,发现10 d时黏附于纳米晶胶原基骨上的骨髓间充质干细胞数明显高于5 d时黏附的细胞数,差异有显著性意义(P<0.01).结论:兔骨髓间充质干细胞与纳米晶胶原基骨体外复合培养结合程度较高.

In vitro co-culture of nano-hydroxyapatite/collagen and mesenchymal stem cells

BACKGROUND: Nano-hydroxyapatite/collagen (NHAC) has similar component, structure, and morphology to natural bone, characterizing by good biocompatibility, multiaperture, large aperture, high porosity, and high pore communication rate; therefore, NHAC is an ideal scaffold material for bone tissue engineering.OBJECTIVE: To explore the feasibility of rabbit bone marrow-derived mesenchymal stem cells (MSCs) and NHAC in constructing tissue-engineered bone.DESIGN, TIME AND SETTING: In vitro observational experiment was performed at Cell Laboratory of Jiangsu University from March to September 2006.MATERIALS: A one-month-old healthy male New Zealand rabbit was provided by Experimental Animal Center of Jiangsu University. NHAC was provided by Material Department of Tsinghua University.METHODS: Rabbit MSCs were isolated and purified in vitro, and the 3rd-generation MSCs were seeded onto NHAC. MAIN OUTCOME MEASURES: ① Morphology and growth curve of rabbit MSCs; ② adhesion grade at day 5 and 10 after culture under scanning electron microscopy; ③ the amount of MSCs that adhered with NHAC on the 5th and 10th days. RESULTS: Rabbit MSCs were grown adherently with great reproductive activity, and the auxodrome was in accordance with Logistic auxodrome. Scanning electron microscopy demonstrated that the amount of MSCs on the 10th day was significantly greater than the amount on the 5th day (P < 0.01).CONCLUSION: The adhesion degree of the compound of MSCs and NHAC is high.