应用兔骨髓基质细胞膜片与珊瑚支架构建组织工程骨

Combined rabbit bone marrow stromal cell-sheet technique and coral scaffold for tissue engineering bone

目的:探讨应用富血小板血浆与骨髓基质细胞(BMSCs)构建骨髓基质细胞膜片,以及应用骨髓基质细胞膜片与珊瑚支架构建组织工程骨的可行性。方法:将取自同一供体兔的BMSCs与富血小板血浆复合后,共同在体外培养10天,构建出BMSCs膜片,用其包裹珊瑚支架后植入裸鼠背部皮下,另以单纯BMSCs种植到珊瑚支架及以单一珊瑚植入作为对照。术后4周和8周取材,通过组织学观察和组织形态测量分析,评价其成骨情况。采用SPSS11.0软件,对所得数据进行统计学分析。结果:BMSCs与富血小板血浆共同在体外培养10d,可构建出厚约2mm的BMSCs膜片。用其包裹珊瑚支架后植入裸鼠背部皮下4周和8周,在珊瑚表面及其孔洞内有大量软骨和骨质形成,其成骨量明显大于单纯BMSCs种植组。单一珊瑚植入组未见骨或软骨形成。结论:将富血小板血浆与BMSCs复合后共同在体外培养,可构建出具有一定厚度的BMSCs膜片,将此膜片包裹珊瑚支架所形成的膜片-支架复合体具有良好的成骨活性。

PURPOSE： To study the feasibility of constructing marrow stromal cell-sheet and tissue engineering bone by combined marrow stromal cell-sheet technique and coral scaffold. METHODS： About 5×10^4 rabbit bone marrow stromal cells （BMSCs） were suspended in 200μl of platelet-rich plasma （PRP）, and the mixture was seeded and cultured on a plate. Marrow stromal cell-sheet was harvested after ten days of culture. The cell-sheets comprising muhilayered BMSCs were wrapped around coral scaffolds, and cell-sheet-scaffold constructs were transplanted under the skin of nude mice. Coral scaffolds loading BMSCs or coral scaffolds alone acted as control. The new bone formation was investigated by histology and histomorphometry 4 and 8 weeks after operation.SPSS 11.0 Software package was used for statistical analysis. RESULTS： New bone and cartilage formation was observed in specimens from both cell-sheet-scaffold constructs group and cell-scaffold constructs group in ectopic sites, and osteogenesis followed the pattern of endoehondral bone formation, with woven hone matrix subsequently maturing into fully mineralized compact bone. Histomorphometric analyses showed enhanced cartilage and bone formation in cell-sheet-scaffold construct group 4 and 8 weeks after transplantation. No bone or cartilage formation could be observed in coral scaffold group. CONCLUSION： Bone tissue similar to native bone may be regenerated utilizing marrow stromal cell-sheet technique in conjunction with coral scaffolds.Supported by Medical Science and Technology Research ＂Eleventh Five Year Project＂of Nanjing Command （Grant No. 06M128）,and Research Fund of Medical Science and Technology Innovation Project of Nanjing Command（Grant No. 07M087）.