Li Jun, Huang Zeyu, Chen Liyan, Tang Xin, Fang Yue, Liu Lei
Department of Orthopaedics, West China Hospital, Sichuan University37# Wainan Guoxue Road, Chengdu 610041, People's Republic of China.
Am J Transl Res. 2017 Jul 15;9(7):3200-3211. eCollection 2017.
The aim of the present study was to investigate the effect of modified heterogeneous deproteinized bone combined with bone marrow mesenchymal stem cells (BMSCs) in the restoration of a validated bone defect model. BMSCs were identified by flow cytometry and multilineage differentiation assay. The structural features of the modified heterogeneous deproteinized bone scaffold and biocompatibility between BMSCs and the scaffold were confirmed by scanning electron microscope (SEM) detection. The cytotoxicity of the modified heterogeneous deproteinized bone scaffolds were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenytetrazolium bromide (MTT) assay. SEM detection proved that modified heterogeneous deproteinized bone scaffold had no negative impact on the proliferation of BMSCs. MTT assay results demonstrated that the scaffold had no apparent cytotoxicity. Biomechanical detection showed that the stiffness and ultimate loading of tibias in the scaffold + BMSCs group were significantly higher than those of the scaffold alone group (P < 0.05) and the control group (P < 0.01). Histological analyses confirmed that the greatest quantity of new bone was generated in the scaffold + BMSCs group, when compared with all other groups, at 8 weeks' post-operation. The bone mineral density (BMD) in the scaffold + BMSC group was significantly higher than that of the scaffold alone group (P < 0.05) and the control group (P < 0.01). Fluorometric analyses confirmed the presence of BMSCs at high concentration within the bone defect areas in the scaffold + BMSCs group at 4 weeks after transplantation. These findings suggest that the modified heterogeneous deproteinized bone scaffold seeded with BMSCs can effectively enhance the restoration of bone defects.
本研究的目的是探讨改良异种脱蛋白骨联合骨髓间充质干细胞（BMSCs）对已验证的骨缺损模型修复的影响。通过流式细胞术和多向分化试验鉴定BMSCs。通过扫描电子显微镜（SEM）检测证实了改良异种脱蛋白骨支架的结构特征以及BMSCs与支架之间的生物相容性。采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐（MTT）法检测改良异种脱蛋白骨支架的细胞毒性。SEM检测证明改良异种脱蛋白骨支架对BMSCs的增殖没有负面影响。MTT试验结果表明该支架没有明显的细胞毒性。生物力学检测显示，支架+BMSCs组胫骨的刚度和极限负荷明显高于单纯支架组（P<0.05）和对照组（P<0.01）。组织学分析证实，与所有其他组相比，术后8周时支架+BMSCs组产生的新骨量最多。支架+BMSC组的骨密度（BMD）明显高于单纯支架组（P<0.05）和对照组（P<0.01）。荧光分析证实，移植后4周，支架+BMSCs组骨缺损区域内存在高浓度的BMSCs。这些发现表明，接种BMSCs的改良异种脱蛋白骨支架可以有效促进骨缺损的修复。
