部分间质血管-脱细胞骨基质-壳聚糖支架在促进兔桡骨骨折愈合中的作用  

Establishment of rabbit radius fracture model and the role of stromal vascular fraction - acellular bone extraceHular matrix - chitosan scaffold in promoting the healing of rabbits

作　　者：邵擎东 汪铮 李宇飞 许天明 孙久一 江峰 吕峰霞 

机构地区：[1]解放军上海第四五五医院骨科,上海200052 [2]解放军上海第四五五医院生物诊疗科,上海200052 [3]解放军上海第四五五医院整形外科,上海200052

出　　处：《中华实验外科杂志》2017年第1期97-101,共5页Chinese Journal of Experimental Surgery

基　　金：全军医药卫生科研项目（14ZD09）

摘　　要：目的制备结构稳定，良好的骨诱导性与生物相容性的部分间质血管（SVF）-脱细胞骨基质（ABECM）-壳聚糖（CS）复合支架，观察SVF-ABECM-CS支架对促进兔桡骨骨折愈合的疗效。方法用Triton X-100、十二烷基硫酸钠（SDS）的脱细胞方法对猪股骨进行脱细胞处理，应用溶液共混、冷冻干燥法制备ABECM-CS复合支架。对SVF-ABECM-CS支架进行定性分析，观察SVF-ABECM-CS复合多孔支架的形态学、孔隙率、力学性能和细胞-支架复合培养的相容性。选用成年健康的新西兰大白兔12只，沿兔前臂桡侧暴露桡骨干并锯断留有0.3 cm的骨折间隙造成骨折模型。在每只大白兔的右侧桡骨骨折间隙处植入制备好的SVF-ABECM-CS复合支架，其左侧为空白对照组。通过X线片、生物力学检测及组织形态学检测等方法对兔桡骨骨折模型两侧桡骨的骨折愈合进行比较。结果ABECM材料苏木素-伊红（HE）染色、Hoechest33258荧光染色均无细胞残留，茜素红染色、Ⅰ型胶原免疫组织化学染色、骨形态发生蛋白-2抗体染色阳性。扫描电镜显示支架具有多孔结构，孔径50～200 μm，孔隙率为（53.50±4.23）%，力学测试支架干燥状态纵向压缩模量为（33.23±14.45） mPa，支架湿润状态下的压缩模量为（2.27±1.13） mPa。细胞-支架复合培养相容性良好。术后第4周和第12周实验组通过X线片反映出的骨愈合结果明显强于对照组；术后第12周生物力学检测实验组中最大负荷、抗弯曲强度下载荷/位移等参数与对照组比较差异均有统计学意义（t＝7.418、5.131、2.838，P＝0.004、0.009、0.027）。而弹性模量实验组与对照组比较提高了13.59%，差异无统计学意义（t＝1.455，P＝0.064）；术后第12周，组织形态学结果显示，实验组成骨及塑形等方面明显优于对照组。结论制备的ABECM-CS复合多孔支架去细胞彻底，保留了脱细胞骨基质主要成分�Objective To prepare stable structure and good bone induction and biological compatibility of stromal vascular fraction （SVF）-acellular bone extracellular matrix（ABECM）-chitosan composite scaffold study SVF-ABECM-chitosan scaffolds to promote the healing of rabbit radius fracture.Methods Using Triton X-100 and sodium dodecyl sulfate （SDS） acellular method of porcine femur were decellularized, using a solution blending, freeze drying method preparing ABECM-chitosan composite scaffolds. The qualitative analysis of SVF-ABECM-chitosan scaffolds, observe SVF-ABECM-chitosan composite porous scaffold morphology, porosity, mechanical properties and cell scaffold composite culture compatibility.12 adult healthy New Zealand white rabbits were selected, and the fracture model was caused by the fracture gap of about 0.3 cm, which was exposed to the radial shaft of the radial side of the forearm of the rabbit. The SVF-ABECM-chitosan composite scaffold was prepared in the right radius fracture space of each animal, and the left side was the blank control group. The fracture healing was compared between the two sides of the radius fracture model of rabbits by X-ray, biomechanical test and tissue morphology.Results ABECM materials were stained with he and hoechest 33258 fluorescence staining showed no residual cells, immunohistochemical staining of alizarin red staining and type I collagen and bone morphogenetic occurred protein-2 antibody staining positive. Scanning election microscopy （SEM） showed that the scaffolds have a porous structure, the aperture 50-200 μm, porosity of （53.50±4.23）%, dry mechanical test support longitudinal compression modulus is （33.23±14.45） mPa, support wet state of compression modulus of （2.27±1.13） mPa. Cell scaffold composite culture has good compatibility. After 4 weeks and 12 weeks in the experimental group by X-ray films reflect the bone healing was stronger than that of the control group. After 12 weeks, the biomechanical test group in the maximum load, anti ben

关 键 词：部分间质血管 脱细胞骨基质 壳聚糖 桡骨骨折 兔 

分 类 号：R318.08[医药卫生—生物医学工程] R683.4[医药卫生—基础医学]