骨髓来源成骨细胞接种于珊瑚颗粒中异位成骨的观察

Inoculation of marrow derived osteoblasts to ectopia osteogenesis in coral granules

目的:为了寻找更好的构建组织工程骨的方法,探讨以天然多孔珊瑚作为支架材料,接种骨髓来源成骨细胞,植入体内后异位再造骨组织的可行性。方法:分离、培养、扩增兔骨髓基质干细胞,细胞长满后用基因重组人骨形成蛋白-2诱导3d;收集细胞,按5×107的浓度接种于珊瑚颗粒中,体外继续培养48h后,植入裸鼠背部皮下组织中,分别于植入术后1,2月取材,通过大体标本、X射线片、组织学检查观察新骨的形成情况。结果:植入术后1,2月,在裸鼠背部皮下组织中均形成了硬组织,1月标本表面粗糙,红白相间,2月标本表面较为光滑,为暗红色;2月标本X线摄片检查可见,在珊瑚颗粒间有明显的X射线阻射影;组织学检查可见,1月标本中,在珊瑚颗粒之间及珊瑚孔隙内,均有大量类骨质形成,并可见较多软骨组织,2月标本中新骨已较为成熟。结论:骨髓来源成骨细胞接种于珊瑚颗粒中,植入体内后可以在异位有效形成骨组织,为其用于组织工程骨提供了有利的实验依据。

AIM: To probe into the feasibility of ectopic reconstructed bone tissues after marrow derived osteoblasts(MOs) inoculated with natural porous coral as brace materials implanted into the body so as to look for a better method of making tissue engineering bone. METHODS: The matrix stem cells of bone marrow in rabbits were isolated, cultured and expanded. The cells were induced by recombinant human bone morphogenetic protein 2 for three days when becoming confluent to improve osteoblastic phynotype. Cells were collected and inoculated to coral granules at the concentration of 5×107, and they were cultured in vitro for 48 hours, and then implanted subcutaniously into nude mice back. The materials were taken and evaluated by gross inspection, X ray examination and histological observation 1 and 2 months after implantation. RESULTS: Hard tissues were formed subcutaniously in nude mice back in 1 and 2 months after implantation. Specimens of 1 month had rough surface of white and red. Specimens of 2 months had smooth surface of glistering deep red. X ray examination showed that there was an obvious new bone formation among the coral granules.Histological observation proved that bone like tissue formed among the space of granules and in the pores of coral,and cartilage tissues could also be observed in some area. In the 2 month specimens, newly formed bone became maturation. CONCLUSION:MOs inoculate to the coral granules, and can effectively form osteogenic tissue in the ectopia after implanted into the body,So it provides the experimental basis for tissue engineered bone.