纳米羟基磷灰石对人脂肪干细胞共混物三维生物打印影响的初步研究

doi:10.3969/j.issn.1671-167X.2016.05.027

北京大学学报(医学版) ›› 2016, Vol. 48 ›› Issue (5): 894-899. doi: 10.3969/j.issn.1671-167X.2016.05.027

纳米羟基磷灰石对人脂肪干细胞共混物三维生物打印影响的初步研究

宋杨，王晓飞，王宇光，董凡，吕培军△

（北京大学口腔医学院·口腔医院,口腔医学数字化研究中心,口腔修复教研室口腔数字化医疗技术和材料国家工程实验室口腔数字医学北京市重点实验室，卫生部口腔医学计算机应用工程技术研究中心，北京100081）

出版日期:2016-10-18 发布日期:2016-10-18
通讯作者: 吕培军 E-mail:kqlpj@bjmu.edu.cn
基金资助:
教育部科学技术研究项目（113002A）资助

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

SONG Yang, WANG Xiao-fei, WANG Yu-guang, DONG Fan, LU Pei-jun△

(Center of Digital Dentistry, Department of Prosthodontics, Peking University School and Hospital of Stomatology & Na-tional 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)

Online:2016-10-18 Published:2016-10-18
Contact: LU Pei-jun E-mail:kqlpj@bjmu.edu.cn
Supported by:
Supported by Science and Technology Research Project of Chinese Ministry of Education, China (113002A)

摘要/Abstract

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，hASCs）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hASCs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/mL）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14 d，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。

关键词: 打印, 三维, 纳米羟基磷灰石, 间充质干细胞, 脂肪组织

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: 1×106/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.

Key words: Printing, three-dimensional, Nano-hydroxyapatite, Mesenchymal stromal cells, Adipose tissue

中图分类号:

R329.4

宋杨，王晓飞，王宇光，董凡，吕培军. 纳米羟基磷灰石对人脂肪干细胞共混物三维生物打印影响的初步研究[J]. 北京大学学报(医学版), 2016, 48(5): 894-899.

SONG Yang, WANG Xiao-fei, WANG Yu-guang, DONG Fan, LU Pei-jun. A preliminary study for the effect of nano hydroxyapatite on human adipose-derived mesenchymal stem cells mixture 3D bio-printing[J]. Journal of Peking University(Health Sciences), 2016, 48(5): 894-899.

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纳米羟基磷灰石对人脂肪干细胞共混物三维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

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