组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2016, Vol. 12 ›› Issue (6): 340-352.doi: 10.3969/j.issn.1673-0364.2016.06.003

• 论著 • 上一篇    下一篇

BMSCs联合3D打印PLLA支架促进兔颅骨PDO的研究

赵丹阳，姜闻博，张海峰，杜朝，杜子婧，韩冬   

  1. 上海交通大学医学院附属第九人民医院整复外科；上海交通大学医学院附属第九人民医院3D打印技术临床转化研发中心
  • 发布日期:2020-07-23

Research of Bone Marrow Mesenchymal Stem Cells Combined with Three-dimensional Printed Poly L-lactic Acid for Promoting Periosteal Distraction Osteogenesis of Rabbit Skull

ZHAO Danyang,JIANG Wenbo,ZHANG Haifeng,DU Chao,DU Zijing,HAN Dong   

  1. Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine;Clinical Translational Research and Development Center of 3D Printing Technology, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine;
  • Published:2020-07-23
  • Contact: 国家自然科学基金项目（81272132,81571944）

摘要： 目的探讨骨髓间充质干细胞(Bone marrow mesenchymal stem cells,BMSCs)促进骨膜牵张成骨(Periosteal distraction osteogenesis,PDO)的可行性。方法全骨髓贴壁培养法获取新西兰白兔BMSCs,体外扩增培养并进行成骨、成脂、成软骨诱导分化。用3D打印(Three-dimensional printing,3D printing)的左旋聚乳酸(Poly l-lactic acid,PLLA)支架牵张兔颅骨骨膜,在牵张期结束时,实验组于骨膜与颅骨间隙内注射兔BMSCs,对照组则注射等量生理盐水(Normal saline,NS)。分别在固定期后的4周和8周,对兔颅骨的牵张部位进行取材,行Micro-CT扫描和组织学染色,评价新骨形成情况。结果全骨髓培养法获取的兔BMSCs具有多向分化能力。Micro-CT显示实验组和对照组的牵张区域在4周和8周均有新骨生成,实验组在两个时间点的新生骨量(Bone volume,BV)、骨量/组织量(Bone volume/Tissue volume,BV/TV)、骨密度(Bone mineral density,BMD)、骨小梁数量(Trabecular number,Tb.N)和厚度(Trabecular thickness,Tb.Th)均明显高于对照组,骨小梁间隔(Trabecular separation,Tb.Sp)小于对照组。HE染色显示,实验组与对照组均有新骨生成,实验组新生骨小梁增厚增多,比对照组成熟,且与颅骨皮质的界线不明显。结论在兔颅骨PDO区域内注射自体BMSCs可以补充成骨细胞的不足,有利于骨膜牵张成骨。

关键词: 骨髓间充质干细胞, 三维打印, 左旋聚乳酸, 骨膜牵张成骨

Abstract: Objective To investigate whether bone marrow mesenchymal stem cells （BMSCs） can promote periosteal distraction osteogenesis （PDO）. Methods BMSCs isolated from New Zealand white rabbits were cultured by the whole bone marrow adherent method and induced to differentiate into osteocytes, adipocytes and chondrocytes in vitro. Poly l-lactic acid （PLLA） scaffold produced by three-dimensional printing （3D printing） technology was used to distract the calvarial periosteum of rabbit. At the end of distraction period, BMSCs were injected into the gap between periosteum and skull in experimental group while the control group received equal normal saline （NS）. The distraction parts of rabbits＂ skull were collected after 4 and 8 weeks of consolidation periods. The newly formed bone of both groups was evaluated by Micro-CT scanning and histological staining. Results BMSCs obtained by the whole bone marrow culture method had the ability of multi-directional differentiation. Micro-CT scanning showed that there were new tissues formed in the distraction area at each time point both in experimental group and control group. The bone volume （BV）, bone volume/tissue volume （BV/TV）, bone mineral density （BMD）, trabecular number （Tb.N） and trabecular thickness （Tb.Th） in experimental group were higher than that in control group, however, the trabecular separation （Tb.Sp） in the experimental group was lower than that of the control group. HE staining also indicated new bone formation both in experimental group and control group. Compared with the control group, the newly formed trabecular bone in the experimental group was thicker and maturer, and the boundary between the new bone and the skull cortex was not obvious. Conclusion Injection of autologous BMSCs into the PDO region of rabbit skull can supplement the deficiency of osteocytes, which is beneficial to PDO.

Key words: Bone marrow mesenchymal stem cells, Three-dimensional printing, Poly l-lactic acid, Periosteal distraction osteogenesis

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