中文摘要：

目的：初步研究纳米羟基磷灰石对人脂肪间充质干细胞（human adipose-derived mesenchymal stem cells,h ASCs）共混物三维生物打印体细胞增殖和成骨分化的影响。方法：以P5代h ASCs作为种子细胞,将10 g/L纳米羟基磷灰石加入细胞-海藻酸钠-明胶共混物（20 g/L海藻酸钠,80 g/L明胶,细胞密度约为1×106个/m L）混合后进行三维生物打印,作为实验组;另将细胞-海藻酸钠-明胶共混物直接进行三维生物打印,作为对照组。在打印后的既定时间点上,对实验组和对照组分别进行显微镜观察、CCK-8检测、Western blot检测和PCR检测,分析打印体细胞增殖能力和成骨分化能力。结果：显微镜观察和CCK-8检测结果显示,实验组和对照组打印体内细胞在打印完成后24 h和7 d均呈现良好的增殖能力。Western blot结果显示,在打印完成后14 d,实验组和对照组的Runt相关转录因子（Runt-related transcription factor 2,RUNX2）蛋白水平差异无统计学意义。PCR结果显示,在打印完成后14d,实验组的成骨相关基因RUNX2、osterix（OSX）和骨钙素（osteocalcin,OCN）表达明显高于对照组。结论：纳米羟基磷灰石加入后,打印体内细胞仍具有良好的增殖性,且打印体的体外成骨分化能力得到了增强。

英文摘要：

Objective： To study the effect of nano hydroxyapatite on human adipose-derived mesenchymal stem cells（ h ASCs） mixture 3D bio-printing for cells＇ proliferation and osteogenesis. Methods： P5 h ASCs were used as seed cells,10 g / L nano hydroxyapatite was added into the cell-sodium alginategelatin mixture（ concentration： 20 g / L sodium alginate,80 g / L gelatin; cell density： 1 × 106/ m L）,then the mixture was printed by 3D bio-printer as the experimental group. And the cell-sodium alginategelatin mixture without nano hydroxyapatite was printed as the control group. Respectively,both the experimental and control groups were detected by microscope,CCK-8,Western blot and PCR at certain time pointsafter being printed,whose cells＇ proliferation and osteogenic differentiation were analyzed.Results： The microscopic observation and CCK-8 results showed that the cells of the experimental group and the control group both had a good proliferation 24 h and 7 d after being printed. The Western blot results showed that 14 d after printing,the expression of Runt-related transcription factor 2（ RUNX2） had no statistical difference between the experimental group and control group. The PCR results showed that14 d after printing,the expression of osteogenesis-related genes（ RUNX2,osterix,and osteocalcin） was significantly higher in the experimental group than in the control group. Conclusion： Nano hydroxyapatite can increase osteogenic differentiation of the h ASCs mixture after bio-printing,in which the cells still have a good proliferation.