摘要
背景:软骨细胞外基质作为一种天然材料,为依赖组织生长的细胞提供了理想的微环境,已成为一种理想的软骨修复支架材料。目的:将人关节软骨来源的细胞外基质制备成软骨组织工程支架,评价其理化性质及生物学性能。方法:收集人源性关节软骨组织,予粉碎成匀浆、去细胞处理、冷冻干燥法制备成软骨组织工程支架。观察:①扫描电镜观察支架形态;②支架的孔隙率、吸水率测定,测量支架力学性质;③支架的成分:使用组织化学和免疫组织化学法染色观察;④支架细胞毒性:用人软骨细胞外基质支架浸提液与常规细胞培养液做对比,培养兔软骨细胞不同时间后使用MTT比色法检测;⑤细胞黏附及增殖情况:使用扫描电镜及共聚焦显微镜观察人软骨细胞与支架共培养后细胞黏附增殖情况,支架细胞复合体切片染色观察。结果与结论:①制备而成的支架呈三维多孔海绵状纵行取向形态,支架孔隙周围有软骨纤维,支架苏木精-伊红染色未见细胞核、番红O及天狼星红染色均证实支架含有胶原和软骨基质成分;②支架的孔隙率为(91.8±2.9)%、吸水率为(93.5±1.4)%;③人软骨细胞外基质支架浸提液经MTT检测,对软骨无细胞毒性;④共培养后,人软骨细胞在支架孔隙周壁上黏附、增殖且分布均匀。结果表明,人软骨细胞外基质多孔取向支架的成分与天然软骨近似,结构可供细胞均匀黏附、增殖良好,组织相容性好,可作为一种组织工程修复软骨缺损的支架材料。
BACKGROUND:Cartilage-derived extracellular matrix as a natural material provides an ideal microenvironment for cells that depend on tissue growth and has become an ideal scaffold material for cartilage repair.OBJECTIVE:To prepare human articular cartilage-derived extracellular matrix as a cartilage tissue engineering scaffold and to evaluate its physicochemical properties and biological properties.METHODS:The human articular cartilage tissue was collected,pulverized into homogenate,decellularized,and freeze-dried to prepare a cartilage tissue engineering scaffold.Scaffold morphology was observed under scanning electron microscope.The porosity and water absorption and mechanical properties of the cartilage tissue engineering scaffold were determined.The composition of tissue engineering scaffold was observed by histochemistry and immunohistochemistry.Rabbit chondrocytes were cultured with the leaching liquor of human cartilage-derived extracellular matrix tissue engineering scaffold for different time periods.Routine cell culture fluid was used as control.The cytotoxicity of tissue engineering scaffold was detected by MTT colorimetry.Human chondrocytes and tissue engineering scaffold were co-cultured and cell adhesion and proliferation were observed under the scanning electron microscope and confocal microscope.The cell-scaffold complex sections were stained by histochemistry and immunohistochemistry.RESULTS AND CONCLUSION:The prepared scaffold had three-dimensional porous sponge-like longitudinally oriented structure.There were cartilage fibers around the scaffold pores.After hematoxylin-eosin staining,no nucleus was observed.Both safranin O and Sirius red staining confirmed that cartilage tissue engineering scaffold contained collagen and cartilage matrix.The porosity and water absorption of the scaffold was(91.8±2.9)%and(93.5±1.4)%,respectively.MTT results showed that the leaching liquor of human cartilage-derived extracellular matrix was non-toxic to chondrocytes.After co-culture,human chondrocytes adhered,p
作者
彭礼庆
罗旭江
张彬
沈师
黄波
高超
鲁晓波
刘舒云
郭全义
Peng Liqing;Luo Xujiang;Zhang Bin;Shen Shi;Huang Bo;Gao Chao;Lu Xiaobo;Liu Shuyun;Guo Quanyi(Department of Bone and Joint Surgery,Affiliated Hospital of Southwest Medical University,Luzhou 646000,Sichuan Province,China;Institute of Orthopedics,First Medical Center,Chinese PLA General Hospital,Beijing Key Lab of Regenerative Medicine in Orthopedics,Key Laboratory of Musculoskeletal Trauma&War Injuries,PLA,Beijing 100853,China)
出处
《中国组织工程研究》
CAS
北大核心
2019年第34期5436-5441,共6页
Chinese Journal of Tissue Engineering Research
基金
国家重点研发计划(2018YFC1105900)，项目参与人：刘舒云
国家自然科学基金(81772319)，项目负责人：郭全义
北京自然科学基金(7172203)，项目负责人：郭全义~~
关键词
组织工程
细胞外基质
支架
软骨
同种异体
tissue engineering
extracellular matrix
scaffold
cartilage
allogenic
