中国组织工程研究2016，Vol.20Issue(46)：6845-6853,9.DOI:10.3969/j.issn.2095-4344.2016.46.001
同种异体骨髓间充质干细胞在比格犬体内异位构建组织工程骨
Ectopic osteogenesis of tissue-engineered bone using allogenic bone marrow mesenchymal stem cells in beagle dogs
摘要
Abstract
BACKGROUND:Al ogenic bone marrow mesenchymal stem cel s as seed cel s for tissue engineering have become the future trend of development. OBJECTIVE:To investigate the osteogenic effects of al ogenic bone marrow mesenchymal stem cel s and the outcome in vivo. METHODS:Bone marrow mesenchymal stem cel s from beagle dogs were marked with chloromethylbenzoyl ammonia fluorescent dye (CM-Dil), and the proliferation of labeled cel s was measured using MTT assay in vitro. Autologous or al ogenic bone marrow mesenchymal stem cel s were inoculated into coral andβ-tricalcium phosphate scaffolds for 7 days osteogenic induction and then subcutaneously implanted into the back of beagle dogs. Dogs undergoing blank scaffold implantation served as negative controls. Hematoxylin-eosin staining was used to observe new bone formation at 3 days, 1, 2, 4, 8, 12 weeks after surgery. Bone formation area was statistical y analyzed using ipp software. In the CM-Dil group, frozen sections were made to trace the in vivo outcome of bone marrow mesenchymal stem cel s under a fluorescence microscope. RESULTS AND CONCLUSION:The osteogenesis speed in the al ogenic bone tissue engineering group was faster than that in the autologous bone tissue engineering group at 4-8 weeks after implantation, but no significant difference between the two groups was found beginning at the 12th week. At 4 weeks after implantation, the expression ofγ-carboxy glutamic acid protein in the autologous bone tissue engineering group was higher than that in the al ogenic bone tissue engineering group, prompting the bone mineralization appeared earlier in the latter group than the former one. ELISA results showed that the expression of alkaline phosphatase and osteocalcin in the autologous bone tissue engineering group was higher than that in the al ogenic bone tissue engineering group at 4 weeks after implantation, and then the expression showed no difference at 12 weeks. CM-Dil labeling results showed that the number of al ogenic bone marrow mesenchymal stem cel s was reduced significantly compared with that of autologous bone marrow mesenchymal stem cel s. Al these findings indicate that the ectopic osteogenesis of the al ogenic tissue-engineered bone in large animals is found within 12 weeks after implantation, but the osteogenesis efficiency at early stage (within 8 weeks) is lower compared with the autologous tissue-engineered bone. This difference may be related to the post-implantation immunoreactions that lead to the reduction in cel number.关键词
组织构建/骨组织工程/骨髓间充质干细胞/组织工程骨/异位成骨/CM-Dil/转归/山东省自然科学基金分类
医药卫生引用本文复制引用
武京国,马燕,曹霏霏,王德峰,张庆富,张喜善,李勇..同种异体骨髓间充质干细胞在比格犬体内异位构建组织工程骨[J].中国组织工程研究,2016,20(46):6845-6853,9.基金项目
山东省自然科学基金(博士基金)(BS2015SW030)；山东省医药卫生科技发展计划项目(2016WS0614)；泰安市科技发展计划项目(2015NS1136)@@@@the Natural Science Foundation of Shandong Province, China, No. BS2015SW030 （博士基金）
the Medical Science and Technology Development Plan of Shandong Province, No.2016WS0614 （）
the Science and Technology Development Plan of Taian City, China, No.2015NS1136 （）
