文章摘要
罗春阳,刘 杨,张 啸,范卫民.应用明胶甲基丙烯酰胺低温3D打印组织工程软骨[J].南京医科大学学报,2020,(4):533~537
应用明胶甲基丙烯酰胺低温3D打印组织工程软骨
Low⁃temperature 3D printing of tissue⁃engineered cartilage with gelatin methacrylamide
投稿时间:2020-01-07  
DOI:10.7655/NYDXBNS20200413
中文关键词: 软骨组织工程  3D生物打印  明胶甲基丙烯酰胺
英文关键词: tissue⁃engineered cartilage  3D bioprinting  gelatin methacrylamide
基金项目:国家自然科学基金(81974332,81672169)
作者单位
罗春阳 南京医科大学第一附属医院骨科江苏 南京 210029 
刘 杨 南京医科大学第一附属医院骨科江苏 南京 210029 
张 啸 南京医科大学第一附属医院骨科江苏 南京 210029 
范卫民 南京医科大学第一附属医院骨科江苏 南京 210029 
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中文摘要:
      目的:探讨应用低温3D打印制作的明胶甲基丙烯酰胺(gelatin methacrylamide,GelMA)支架能否构建高质量组织工程软骨。方法:经预冷处理后,利用低温沉积3D打印技术制备低浓度GelMA支架。大体观察、扫描电镜观察支架宏观、微观结构,并对水凝胶进行流变学和生物力学检测,以DMEM培养液培养大鼠骨髓间充质干细胞作为对照,细胞增殖(CCK?8法)和细胞活性(Live/Dead染色法)检测支架生物相容性,组织学染色定性分析支架成分。裸鼠皮下包埋支架,苏木素?伊红(HE)染色、阿利新蓝染色、番红染色评估软骨细胞外基质结构。结果:大体观察和扫描电镜观察示5%GelMA支架孔径均一,结构规整。与纯GelMA相比,添加细胞的支架力学强度稍强。骨髓间充质干细胞能在支架上黏附并增殖。体内体外组织学染色均表明构建的组织工程软骨在大小、色泽、氨基葡糖聚糖分泌等方面均满足组织工程软骨要求。结论:5%GelMA生物墨水可以在低温条件进行3D打印并成功构建组织工程软骨。
英文摘要:
      Objective:To investigate whether gelatin methacrylamide(GelMA)scaffolds made by low?temperature 3D printing could construct high?quality tissue?engineered cartilage. Methods:After pre?cooling,low?temperature deposition 3D printing technology was used to prepare low?concentration gelatin methacrylamide scaffolds. Macroscopic and scanning electron microscopic observation of the macroscopic and microscopic structure of the scaffold were followed by a rheological and biomechanical detection of the hydrogel. Cells were cultured with DMEM as control,cell proliferation(CCK?8 method)and cell activity(Live/Dead staining method)were used to detect the biocompatibility of the scaffold,and histological staining was used to qualitatively analyze the scaffold components. Nude mice were scaffolded subcutaneously for hematoxylin and eosin(HE)staining,alcian blue staining,and safranin staining to highlight the extracellular matrix structure of the cartilage. Results:Macroscopic and scanning electron microscopic observations showed that the 5% GelMA scaffolds had uniform pore diameters and a regular structure. The results of CCK?8 and live/dead cell staining showed that the scaffolds were biocompatible,and the in vivo and in vitro histological staining showed that the constructed tissue?engineered cartilage met the requirements in colors,glycosaminoglycan(sGAG)secretion and other aspects. Conclusion:5% GelMA bio?ink can be used for 3D printing at low temperature and to successfully construct tissue?engineered cartilage.
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