构建大段可控微结构组织工程骨的实验研究  

An experimental study on composition of segmental tissue engineered bone with controllable microstructure

作　　者：王林[1] 王臻[1] 李祥[2] 李涤尘[2] 桑宏勋[1] 郭征[1] 

机构地区：[1]第四军医大学西京医院全军骨科研究所,西安710032 [2]西安交通大学先进制造研究所,西安710049

出　　处：《中国骨与关节外科》2008年第3期210-216,共7页Chinese Journal of Bone and Joint Surgery

基　　金：国家自然科学基金(50235020)

摘　　要：目的应用仿生设计和光固化快速成型间接构造方法构建大段可控微结构组织工程骨,观察其与成骨细胞在体外培养条件下的生物相容性。方法基于对密质骨哈佛系统的简单仿生,应用计算机辅助设计(CAD)软件设计具有微丝架结构的圆柱形支架模型。建模后输入光固化快速成型机,制成树脂模具,充填磷酸钙骨水泥(CPC),800℃下40 min熔去模具,制成CPC支架。将兔颅骨成骨细胞与支架复合培养3、7、14d后,于共聚焦激光扫描显微镜(CLSM)、扫描电镜下观察细胞在支架表面及微孔道内部的黏附、生长和增殖情况。结果支架CAD模型为直径16 mm,长10 mm的圆柱体,内部由5层微丝网平行叠加形成相互连通的三维网架,微丝直径分别为200μm和500μm。制成支架的大体形态及内部三维微结构与设计模型一致,所构成的微孔道孔径范围260～580μm。CLSM下观察:特异性荧光探针标记显示细胞骨架成网状贯穿整个细胞和细胞突起;SEM下观察:培养3d时支架表面有梭型细胞铺展生长,有多个突起,大小10～30μm;7d后细胞在支架表面基本汇合,并向微孔内爬行生长:14d后大量细胞爬行入微孔内。结论基于哈佛系统简单仿生设计和快速成型技术制备的大段组织工程骨,实现了微结构的完全精确调控,其微孔利于成骨细胞进入,具有良好的生物相容性,为大段组织工程骨的仿生制造提供了实验依据。Objective To investigate the composition of a segmental tissue engineered bone with controllable microstructure in an approach of bionic design and stereolithography rapid prototyping indirect fabrication,and observe the biocompatibility of the tissue engineered bone with osteoblasts in vitro culture.Methods Based on simple bionics of Haversian system in cortical bone,The model of cylindrical scaffold with micro-thread net was designed by means of computer aided design(CAD) software.The data of CAD model was imported into stereolithography rapid prototyping machine to get resin mold and then The scaffold was achieved by sintering the mold filled with Calcium Phosphate Cement(CPC) at 800℃for 40 minutes.Osteoblasts were isolated from skull of rabbit and seeded onto CPC scaffolds.3,7 and 14 days after seeding of incubation respectively,adhesion,growth,and proliferation of osteoblasts on scaffolds and within micropore were observed by confocal laser scanning microscopy and scanning election microscopy.Results The CAD model was a cylinder with a diameter of 16mm and a length of 10mm.There was a 3D-net with 5 layers of micro-thread net overlapped within the cylinder.The micro-thread size was 200μm or 500μm.The gross shape and 3D microstructure of the scaffold were achieved in accord with CAD designed model.The arrange of achieved micropore size was 260μm～580μm. In CLSM observation after 3 days of incubation,retiform cytoskeleton in cells marked by fluorescence probe ranged through the whole cell or cellular prominences.In SEM observation after 3 days of incubation,cells exhibited spindle-like morphology and had a diameters of 10～30μm,and after 7 days of incubation,confluent cell layer was observed on scaffolds, and cells were found to grew into the micropores.Furthermore,a great lot of cells grew into the micropores after 14 days of incubation.Conclusion The controllable microstructure of this segmental tissue engineered bone was achieved Completely and accurately in an approach of bionic design and stereoli

关 键 词：微结构 快速成形 羟基磷灰石类 成骨细胞 

分 类 号：R318[医药卫生—生物医学工程]