摘要
目的探索壳聚糖-同种异体骨粉复合多孔支架材料修复骨缺损以及作为骨组织工程支架材料的可行性。方法取壳聚糖与脱钙处理后的SD大鼠同种异体骨粉,按质量比1∶5比例混合后,通过冷冻干燥技术制备复合多孔支架材料,以单纯壳聚糖制备的支架作为对照。大体观察两种支架,乙醇替代法测量孔隙率,扫描电镜测量孔径。于40只SD大鼠双侧股骨内上髁制备直径为3.5 mm的孔,左侧孔内植入复合多孔支架（实验组）,右侧植入单纯壳聚糖支架（对照组）。术后2、4、8、12周取材行大体观察、组织学及免疫学化学染色观察。结果经冷冻干燥制备的复合多孔支架色泽微黄,质地较脆,易折断;支架孔隙孔径以200-300μm为主,孔隙率为76.8%±1.1%,与单纯壳聚糖支架（78.4%±1.4%）比较,差异无统计学意义（t=-2.10,P=0.09）。支架植入大鼠体内后,大体及组织学观察示,随时间延长可见新生骨逐渐填充缺损区,且实验组明显多于对照组;术后4、8、12周实验组单位视野成骨面积均显著大于对照组,比较差异有统计学意义（P〈0.05）。免疫组织化学染色观察示,实验组各时间点均见骨保护素阳性表达,且阳性表达量均显著高于对照组,比较差异有统计学意义（P〈0.05）。结论壳聚糖-同种异体骨粉复合多孔支架材料具有适宜孔隙率和良好成骨活性,可有效修复大鼠骨缺损,其成骨量及成骨速度优于单纯壳聚糖支架,有望作为骨组织工程支架材料。
Objective To explore the feasibility of chitosan/allogeneic bone powder composite porous scaffold as scaffold material of bone tissue engineering in repairing bone defect. Methods The composite porous scaffolds were prepared with chitosan and decalcified allogeneic bone powder at a ratio of 1 ∶ 5 by vacuum freeze-drying technique. Chitosan scaffold served as control. Ethanol alternative method was used to measure its porosity, and scanning electron microscopy（SEM） to measure pore size. The hole of 3.5 mm in diameter was made on the bilateral femoral condyles of 40 adult Sprague Dawley rats. The composite porous scaffolds and chitosan scaffolds were implanted into the hole of the left femoral condyle（experimental group） and the hole of the right femoral condyle（control group）, respectively. At 2, 4, 8, and 12 weeks after implantation, the tissues were harvested for gross observation, histological observation, and immunohistochemical staining. Results The composite porous scaffold prepared by vacuum freeze-drying technique had yellowish color, and brittle and easily broken texture; pore size was mostly 200-300 μm; and the porosity was 76.8%±1.1%, showing no significant difference when compared with the porosity of pure chitosan scaffold（78.4%±1.4%）（t= —2.10, P=0.09）. The gross observation and histological observation showed that the defect area was filled with new bone with time, and new bone of the experimental group was significantly more than that of the control group. At 4, 8, and 12 weeks after implantation, the bone forming area of the experimental group was significantly larger than that of the control group（P〈0.05）. The immunohistochemical staining results showed that osteoprotegerin（OPG） positive expression was found in the experimental group at different time points, and the positive expression level was significantly higher than that in the control group（P〈0.05）. Conclusion Chitosan/allogeneic bone powder composite porous scaffold has suitable porosity and good
出处
《中国修复重建外科杂志》
CAS
CSCD
北大核心
2016年第3期298-302,共5页
Chinese Journal of Reparative and Reconstructive Surgery
关键词
骨组织工程
支架材料
壳聚糖
同种异体骨
大鼠
Bone tissue engineering
Scaffold material
Chitosan
Allogeneic bone
Rat
