低温沉积3D打印负载基质细胞衍生因子1的分级多孔软骨复合支架  

Preparation of hierarchical porous cartilage composite scaffolds loaded with stromal cell-derived factor-1 using low-temperature deposition 3D printing

作　　者：吴江 殷瀚 严子能 田广招 丁振罡 袁勋 付力伟 田壮 眭翔 刘舒云 郭全义 Wu Jiang;Yin Han;Yan Zineng;Tian Guangzhao;Ding Zhengang;Yuan Xun;Fu Liwei;Tian Zhuang;Sui Xiang;Liu Shuyun;Guo Quanyi(Department of Orthopedics,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou Province,China;Institute of Orthopedics,First Medical Center,Chinese PLA General Hospital,Beijing Key Laboratory of Orthopedic Regenerative Medicine,Key Laboratory of Musculoskeletal Trauma&War Injuries,PLA,Beijing 100853,China)

机构地区：[1]贵州医科大学附属医院骨科,贵州省贵阳市550004 [2]中国人民解放军总医院第一医学中心骨科研究所,骨科再生医学北京市重点实验室,全军骨科战创伤重点实验室,北京市100853

出　　处：《中国组织工程研究》2023年第30期4776-4782,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家重点研发计划课题(2019YFA0110600),项目负责人:郭全义。

摘　　要：背景:基于原位组织工程再生技术的飞速发展,促进体内干细胞募集策略为软骨损伤修复提供了新的方向。目的:通过低温沉积3D打印技术构建负载基质细胞衍生因子1的分级多孔细胞外基质支架。方法:采用化学法制备脱细胞软骨细胞外基质。制备脱细胞软骨细胞外基质匀浆,并加入基质细胞衍生因子1,作为生物墨水,采用低温沉积3D打印技术构建分级多孔细胞外基质支架,通过大体观及扫描电镜观察支架的微观结构,通过CCK-8、死活染色及鬼笔环肽骨架染色评价支架的生物相容性,检测支架的体外缓释性能,通过Transwell实验检测支架对骨髓间充质干细胞迁移的影响。结果与结论:①扫描电镜显示,打印的细胞外基质支架呈现分级多孔的结构,孔径分布均匀,纤维交错排列,孔与孔之间相互贯通;②CCK-8实验显示,细胞外基质支架无明显的细胞毒性;死活染色显示,骨髓间充质干细胞能够很好地在细胞外基质支架上生长;鬼笔环肽骨架染色显示,骨髓间充质干细胞在细胞外基质支架上能够很好地铺展开,呈梭形;③细胞外基质支架具有良好的缓释性能,在前7 d内较快速地释放基质细胞衍生因子1,累计释放率达约60%,1-4周释放细胞因子速度逐渐减缓,4-6周的释放规律类似于零级释放动力学,42 d累计释放率约达80%,并且仍有缓慢释放的趋势;④Transwell迁移实验显示,基质细胞衍生因子1可提升细胞外基质支架募集干细胞的能力;⑤结果表明,基质细胞衍生因子1可提高低温沉积3D打印细胞外基质支架募集骨髓间充质干细胞的能力。BACKGROUND:The treatment regimen based on the promotion of stem cell recruitment to achieve in situ regeneration and repair of cartilage damage is a new strategy in cartilage tissue engineering.OBJECTIVE:To prepare hierarchical porous cartilage composite scaffolds loaded with stromal cell-derived factor-1(SDF-1)using low-temperature deposition 3D printing.METHODS:The cartilage was decellularized by a chemical method.The bioink was prepared with the addition of SDF-1 to decellularized cartilage extracellular matrix homogenate.The graded porous extracellular matrix scaffolds were constructed using low-temperature deposition 3D printing.The microstructure of scaffolds was observed by scanning electron microscopy.The biocompatibility of the scaffolds was evaluated by CCK-8,live-dead cell staining,and ghost pen peptide cell skeleton staining.The in vitro sustained release of the scaffolds was tested,and the effects of the scaffolds on the migration of bone marrow mesenchymal stem cells were detected by Transwell assay.RESULTS AND CONCLUSION:(1)Scanning electron microscopy showed that the printed extracellular matrix scaffolds had a hierarchical porous structure,with fibers staggered and interconnected between pores.(2)The CCK-8 results indicated that the prepared extracellular matrix scaffolds had no obvious cytotoxicity.Live-dead cell staining showed that bone marrow mesenchymal stem cells were able to grow well on the extracellular matrix scaffolds;ghost pen peptide cell skeleton staining showed that bone marrow mesenchymal stem cells were able to spread well on the extracellular matrix scaffold in a shuttle shape.(3)The extracellular matrix scaffold had a good sustained release property.SDF-1 was released rapidly during the initial 7 days,with a cumulative release rate of about 60%.The release rate of SDF-1 gradually slowed down during 1-4 weeks,and the release pattern during 4-6 weeks was similar to the zero-order release kinetics,and the cumulative release rate reached about 80%at 42 days,and there was still a tr

关 键 词：细胞外基质 支架 基质细胞衍生因子1 低温沉积3D打印 软骨损伤 骨髓间充质干细胞 迁移 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学] R-331