纳米羟基磷灰石对人脂肪间充质干细胞混合物3D生物打印影响的初步研究

[A preliminary study for the effect of nano hydroxyapatite on human adipose-derived mesenchymal stem cells mixture 3D bio-printing].

作者信息

Song Y, Wang X F, Wang Y G, Dong F, Lv P J

机构信息

Center of Digital Dentistry, Department of Prosthodontics, Peking University School and Hospital of Stomatology & National Engineering Laboratory for Digital and Material Technology of Stomatology & Beijing Key Laboratory of Digital Stomatology, Research Center of Engineering and Technology for Digital Dentistry of Ministry of Health, Beijing 100081, China.

出版信息

Beijing Da Xue Xue Bao Yi Xue Ban. 2016 Oct 18;48(5):894-899.

PMID:27752177

Abstract

OBJECTIVE

To study the effect of nano hydroxyapatite on human adipose-derived mesenchymal stem cells(hASCs) mixture 3D bio-printing for cells' proliferation and osteogenesis.

METHODS

P5 hASCs were used as seed cells, 10 g/L nano hydroxyapatite was added into the cell-sodium alginate-gelatin mixture (concentration: 20 g/L sodium alginate, 80 g/L gelatin; cell density: 1×10/mL), then the mixture was printed by 3D bio-printer as the experimental group. And the cell-sodium alginate-gelatin 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 that 14 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 hASCs mixture after bio-printing, in which the cells still have a good proliferation.

摘要

目的

研究纳米羟基磷灰石对人脂肪间充质干细胞（hASCs）混合三维生物打印中细胞增殖和成骨的影响。

方法

以第5代hASCs为种子细胞，向细胞-海藻酸钠-明胶混合物（浓度：海藻酸钠20 g/L、明胶80 g/L；细胞密度：1×10/mL）中加入10 g/L纳米羟基磷灰石，然后用三维生物打印机打印该混合物作为实验组。将不含纳米羟基磷灰石的细胞-海藻酸钠-明胶混合物打印作为对照组。打印后在特定时间点分别对实验组和对照组进行显微镜观察、CCK-8检测、蛋白质免疫印迹法（Western blot）和聚合酶链反应（PCR）检测，分析细胞增殖和成骨分化情况。

结果

显微镜观察和CCK-8检测结果显示，实验组和对照组细胞在打印后24小时和7天时增殖良好。蛋白质免疫印迹法结果显示，打印后14天，实验组和对照组之间的Runx相关转录因子2（RUNX2）表达无统计学差异。PCR结果显示，打印后14天，实验组中成骨相关基因（RUNX2、osterix和骨钙素）的表达明显高于对照组。