壳聚糖/羟基磷灰石支架修复骨软骨缺损的实验研究  

Repair of articular osteochondral defects with CS/HA bilayered scaffold in rabbits

作　　者：王文良[1] 张华亮 关静[3] 初殿伟[1] 

机构地区：[1]武警医学院附属医院骨科,天津300162 [2]武警九江市支队卫生队,江西九江332000 [3]军事医学科学院卫生装备研究所,天津300162

出　　处：《中国矫形外科杂志》2008年第20期1579-1583,共5页Orthopedic Journal of China

基　　金：国家自然科学基金(项目编号:30740089);武警医学院科研基金(项目编号:WBS2007-13)

摘　　要：[目的]探讨双层壳聚糖（chitosan CS）/羟基磷灰石复合支架（hydroxyapatite HA）修复兔骨软骨缺损的可行性。[方法]采用冻干法和烧结法制作双层壳聚糖（CS）/羟基磷灰石（HA）复合支架,以骨髓间充质干细胞为种子细胞,运用纤维蛋白胶种植技术,以双层壳聚糖（CS）/羟基磷灰石（HA）复合支架为载体,修复骨软骨缺损,实验分3组,A组：BMSc＋支架,B组：单纯支架,C组：未处理。将修复材料植入骨软骨缺损模型,分别于6、12周取材,进行大体观察,组织学检测,改良Wakitani法评分,经统计学处理,比较各组修复效果差异（P〈0.05）。[结果]（1）CS/HA支架CS层孔隙率为76%±5.01%,孔径为200～400μm,平均为300μm左右,孔相通性好,HA层孔隙率为72%±4.23%,孔径为200～500μm,平均为350μm左右,孔相通性好,结合部结合好;（2）P2骨髓间充质干细胞较纯,扫描电镜观察MSCs附着在复合支架上。大体观察和组织学检测显示,A组基本修复软骨缺损,骨缺损有骨小梁长入。B、C组骨软骨缺损修复不良,组织学检测以纤维性组织或无新生组织形成,软骨及骨缺损均明显存在,改良Wakitani评分显示A组在6周、12周2个时间点的各项评分结果,均优于B、C组,且差异有统计学意义（P〈0.05）。[结论]双层壳聚糖（CS）/羟基磷灰石（HA）复合支架可作为骨软骨组织工程支架,结合BMSc可修复软骨与骨的缺损,重建关节的解剖结构和功能。[ Objective ] To evaluate feasibility for the treatment of osteochondral defects on the femoral condyles by chitosan/ hydroxyapatite bilayered scaffold in rabbit knee joint. [ Method] Bilayered scaffolds were produced with chitosan and hydroxyapa- rite using a lyophilization and sintering method. The pore size and interpores of the scaffolds were observed by scanning electron microscopy（SEM） anhydrous ethanol substitution method to determinate its porosity . Rabbit bone mesenchymal stem cells （BM- SC） inoculated onto the scaffold. By SEM scanning,the condition of the cells adhering onto the scaffold was observed. The cell- scaffold composite were implanted into an rabbit model of osteochondral defect 10 hour later. The result was observed 6 and 12 weeks later by HE staining and toluidine blue staining. [ Result ] Physicochemical characterization of CS/HA bilayered scaffold： HA possessed a porosity of 72% ± 4.23% CS layer possessed a porosity of 76% ± 5.0% . SEM microgTaph showed CS pore at the typical transversal cross-section of CS layer with open pores and pore size with average 300 μm ranging from 200 - 400 μm and HA pore at the typical transversal cross-section of HA layer with open pores and pore size with average 350 μm ranging from 200 - 500 μm. The FTIR data showed typical peaks of the CS and HA peaks and no typical peaks of polyethylene glycol peaks. It was observed by SEM that BMSCs adhered on the chitosan scaffold and grew well but did not completely extend , adhered on the HA scaffold and extended partially after BMSCs was implanted on the CS/HA scaffold 10 hours. The treatment of simple CS/HA scaffold improved defect filling compared with that left untreated, while the regenerated tissue was mainly fibrocartilage and showed little bone formation. When implanted with CS/HA scaffold combined with MSCs, most of the defects were filled with a well-established layer of cartilage tissue with a little bone formation observed by HE staining and toluidine blue staining. Modified Wakitani gradi

关 键 词：壳聚糖/羟基磷灰石复合支架 组织工程 骨髓间充质干细胞 骨软骨缺损 

分 类 号：R687.3[医药卫生—骨科学]