组织工程技术仿生构建颅颌骨组织的研究  

Bionic fabrication of cranio-maxillofacial bony implants by tissue engineering techniques

作　　者：姚晖[1] 李俊杰[2] 濮礼臣[3] 张洪杰[4] 董晓燕[2] 张林朴[1] 陈亦平[2] 王冠华[1] 赵峰[2] 

机构地区：[1]天津市口腔医院组织工程实验室,天津市300041 [2]天津大学材料学院高分子材料研究所,天津市300073 [3]天津市整形外科医院,天津市300041 [4]天津市口腔医院颌面外科,天津市300041

出　　处：《中国组织工程研究与临床康复》2007年第40期8047-8051,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：天津市自然科学基金资助项目(003609011)~~

摘　　要：目的:探索构建组织工程化仿生骨种植体的方法流程,并制备成骨细胞-可吸收载体种植体样品,同时尝试建立组织工程化非承载骨种植体的评价方法。方法:实验于2001-05/2005-12分别在天津市口腔医院组织工程实验室和天津大学材料学院高分子材料研究所完成。①通过相分离技术制备壳聚糖/明胶三维网络多孔支架,在支架材料表面原位沉积纳米级的羟基磷灰石晶体,构筑纳米羟基磷灰石/壳聚糖/明胶仿生骨组织工程支架材料,并进行表征和性能检测。②用酶消化法和条件培养法分离、诱导培养中国小型猪成骨细胞作为组织工程种子细胞。③用静态复合共培养法体外构建2种骨组织工程种植体样品:成骨细胞-纳米羟基磷灰石/壳聚糖/明胶仿生骨种植体,成骨细胞-纳米羟基磷灰石/胶原种植体。④采用扫描电镜、透射电镜、FDA荧光、LDH、MTT等定期观测仿生骨样品中细胞形态、细胞增殖速率、碱性磷酸酶活性、矿化结节形成等指标,以比较样品的细胞增殖活性和成骨活性。结果:①成功构筑了具有良好的生物相容性和力学相容性的纳米羟基磷灰石/壳聚糖/明胶,这种材料具有适于细胞黏附与生长的(90±1)%的孔隙率,孔径为100￣300μm的微孔结构,且原位沉积的纳米羟基磷灰石晶体的粒径为50nm左右,接近与天然骨的组成。②自中国小型猪腿骨成功分离培养了成骨细胞,并在诱导培养条件下,表现出很强的增殖活力和成骨活性,适合作为实验用骨组织工程的种子细胞。③成功构建了两种成骨细胞-可吸收载体种植体样品:经检验仿生构建的小型猪成骨细胞-纳米羟基磷灰石/壳聚糖/明胶种植体具有细胞亲和性和体外成骨活性。结论:①在体外成功仿生构建了结构与活性接近天然骨的骨组织工程种植体--纳米羟基磷灰石/壳聚糖/明胶种植体。②初步建立了仿生组织工程化非承载�AIM： To investigate the feasibility of bionic fabrication of oral-maxillofacial and cranio-maxillofacial bony implants by tissue engineering techniques, prepare the sample for osteoblast-absorbable vector implants, and establish the evaluation method of non-bearing bony implants by tissue engineering. METHODS： From May 2001 to December 2005, the experiment was carried out in the Laboratory of Tissue Engineering Technique in Tianjin Stomatological Hospital and the Institute of Polymer Science in Tianjin University Materials Department.① Three-dimensional porous chitosan/glutin network scaffolds were prepared by phase separation techniques, and then nano-hydroxyapatite particles were deposited onto the surface of the scaffolds to develop nano-hydroxyapatite/chitosan/glutin （nHCG） bionic scaffolds. The properties and superficial syndrome of the nHCG scaffolds were estimated. ②By enzyme digestion and conditional culturing, osteoblasts isolated from Chinese mini-pig legs were cultured as bony tissue engineering seeding cells. ③Two types of tissue engineered bony implant samples were fabricated by static co-culture of osteoblasts with nHCG and nano-hydroxyapatite/collagen.④This two types of bony implants were contrasted by observing cell morphous, proliferation rate, alkaline phosphatase activities and mineraliztion node staining with scanning electron microscope, transmission electron microscope, FDA fluorescence, lactate dehydrogenase and MTT methods to determine the osteogenesis activities among 3 types of tissue engineering bony implants. RESULTS： ①The nHCG scaffolds were successfully fabricated and had excellent biocompatibilities and good mechanical properties. The porosity of scaffolds was （90±1）% and the dispersity of apertures was 100-300 μm, which were proper for osteoblasts attaching and distributing; the particle diameter of nano-hydroxyapatite was about 50 nm that was similar to the architecture of natural bone. ②The osteoblasts isolated from Chinese mini-pig were cultured succe

关 键 词：组织工程 纳米材料 仿生 构建 成骨细胞 

分 类 号：R318[医药卫生—生物医学工程]