摘要
背景:补骨脂素是属于植物类雌激素,有大量研究证实其具有促进细胞增殖分化的作用。目的:应用补骨脂素、兔骨内膜间充质干细胞和聚己内酯支架构建人工复合骨,探讨其治疗兔骨不连的效果。方法:①将第3代兔骨内膜间充质干细胞接种于细胞培养板中,分别加入10^-8,10^-7,10^-6mol/L补骨脂素、50mg/L骨形态发生蛋白2进行干预,对照组加入同体积的无菌纯净水。干预第3,5,7天采用MTT法检测细胞增殖情况;②将聚己内酯三维支架加入到细胞培养板底部,兔骨内膜间充质干细胞按1×10^3/孔的量接种于支架上,然后加入10^-6mol/L补骨脂素,联合培养21d后取出即为人工复合骨;③27只新西兰大白兔随机均分为3组,建立桡骨骨不连模型,实验组植入人工复合骨,支架组单纯植入支架,对照组不植入任何材料。术后2,4,8周行病理学苏木精-伊红染色,观察成骨情况。术后第4周拍摄X射线片,观察骨不连愈合情况。结果与结论:①3种浓度补骨脂素均有诱导兔骨内膜间充质干细胞增殖的作用,与对照组比较,10-6mol/L补骨脂素对兔骨内膜间充质干细胞刺激作用最强(P<0.05),而且10-6mol/L补骨脂素组与阳性对照骨形态发生蛋白2组相比差异无显著性意义(P>0.05);②新西兰大白兔桡骨中段骨不连组织苏木精-伊红染色结果显示,实验组成骨细胞数目明显多于支架组和对照组(P<0.05);③X射线片结果显示,实验组骨不连已经愈合,支架组骨折部分愈合,对照组骨不连未愈合;④结果表明,补骨脂素对兔骨内膜间充质干细胞增殖作用与浓度有一定的相关性。补骨脂素可结合兔骨内膜间充质干细胞及聚己内酯支架形成人工复合骨,通过动物实验证实其治疗骨不连效果较好。
BACKGROUND:Psoralen is a plant estrogen,and a large number of studies have confirmed its role in promoting cell proliferation and differentiation.OBJECTIVE:To construct a cell-scaffold composite bone using psoralen,rabbit endosteal mesenchymal stem cells and polycaprolactone and to explore its effect on the treatment of rabbit nonunion.METHODS:(1)Rabbit endosteal mesenchymal stem cells were cultured and cultured until the third generation for each experiment.Passage 3 cells were seeded onto culture plates containing 50 mg/L bone morphogenetic protein 2(positive control),10^-8,10^-7 and 10^-6 mol/L psoralen(low-,middle-,and high-concentration psoralen groups)or the same volume of purified water(control group).The cell proliferation of each group was detected on the 3rd,5th and 7th days after intervention using MTT method.(2)The three-dimensional polycaprolactone scaffold was added to the bottom of the cell culture plate,and rabbit endosteal mesenchymal stem cells were seeded into the scaffold at a density of 1×10^3 per well.Then,10^-6 mol/L of psoralen was added.Cell-scaffold composite bone was taken after 21 days of culture.(3)Animal models of radial nonunion were established in 27 New Zealand white rabbits,and were then randomized into experimental,scaffold and control groups followed by implantation of cell-scaffold composite bone,simple scaffold,and nothing,respectively.Pathological hematoxylin-eosin staining for observation of bone healing was performed at the 2nd,4^th,and 8^th weeks after surgery.Healing of nonunion was observed on the X-ray films that were taken at the 4th week after surgery.RESULTS AND CONCLUSION:(1)Three concentrations of psoralen could induce the proliferation of rabbit endosteal mesenchymal stem cells.Compared with the control group,10^-6 mol/L psoralen exerted the strongest stimulation effect on rabbit endosteal mesenchymal stem cells(P<0.05),whereas there was no significant difference between high-concentration psoralen group positive control group(P>0.05).(2)Pathological hematoxylin
作者
赖丽金
莫浩轩
Lai Lijin;Mo Haoxuan(The People’s Hospital of Longhua,Shenzhen 518109,Guangdong Province,China;Hong Kong Federation of Trade Unions Workers’ Medical Clinics,Hong Kong Special Administrative Region 810015,China)
出处
《中国组织工程研究》
CAS
北大核心
2020年第1期40-44,共5页
Chinese Journal of Tissue Engineering Research
关键词
补骨脂素
兔骨内膜间充质干细胞
细胞增殖
细胞支架
骨不连
psoralen
rabbit endosteal mesenchymal stem cells
cell proliferation
cell scaffold
bone nonunion
