3D生物打印构建复层尿道组织的初步研究

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3D生物打印构建复层尿道组织的初步研究

皮庆猛;王晓磊;张志亮;周碧玉;任晓芸;陆毅;黄一雄;傅敏刚;范志宏

【期刊名称】《组织工程与重建外科杂志》 【年(卷),期】2018(14)5

【摘 要】Objective To investigate the feasibility of multiple-layers

reconstruction of urethral tissue with 3D bioprinting technology. Methods Mixed blend of 7% GelMA and 2% Alginate with green and read microbeads respectively, or 2×106cells/mL urethral epidermal cells and smooth muscle cells, printed on the system of co-axial multiple channel bioprinting platform. The speed of inner and outer layer printing was 10-20 μL/min and CaCl2extruding was 80-200 μL/min. Then the printed structure was observed under microscopy, and the printed hollow tissues loaded urethral cells were tested with live/dead and F-actin staining from day 0 to day 7. The printed hollow structure was perfused with color medium after cultured 24 hours in vitro. Results With hydrogels mixed with microbeads, two distinct sublayers of printed hollow structure can be observed clearly under microscopy. The diameter of inner and outer layer was (663.66±51.74) μm and (976.84±63.43) μm respectively, and wall length of inner and outer layer was (62.06±10.45) μm and (93.78±10.25) μm respectively. The viability of printed urethral tissue was 84.52% and 86.26% at day 0 and day 7. F-actin expression were observed on day 7 after immunofluorescent staining and showed the spread cells. During the


process of perfusion of hollow tissue, colored medium can circulate the channel smoothly without color exuding. Conclusion Urethral hollow structure can be printed with combined hydrogel by the method of co-axial multiple channel bioprinting system. And the urethral cells can survive for long term after printing.%目的 探讨利用生物打印体外构建具有复层结构的尿道空腔组织的可行性.方法 将7%GelMA和2%Algi-nate制备复合水凝胶,分别同绿色和红色的microbeads或2×106cells/mL尿道上皮细胞和尿道平滑肌细胞混匀,通过本实验室改进的同轴多通道打印系统,以10~20 μL/min的速度推进内、外层,以80~200 μL/min的速度推进CaCl2,边打印边交联;打印后不含细胞的空腔结构,置于荧光显微镜下观察;打印后含有细胞的组织,以尿道上皮细胞和尿道平滑肌细胞混合培养液于六孔板内体外培养,分别于第0~7天live/dead染色检测细胞活力,F-actin染色检测细胞铺展情况;将培养24 h的打印后组织,予以红色颜料灌注.结果 复合水凝胶混合microbeads后,荧光显微镜下可以清楚看到内、外双层结构,复层管腔的内、外径分别为(663.66±51.74) μm 和(976.84±63.43) μm,内外亚层壁厚分别为(62.06± 10.45) μm和(93.78±10.25) μm.尿道细胞在第0天和第7天的细胞活力分别为84.52%和86.26%,免疫荧光染色第7天F-actin染色铺展均匀,提示细胞铺展生长.灌注过程中,打印后空腔组织内红色灌注液可以顺利通过管腔结构,颜料并无外溢,管壁无破损.结论 利用复合水凝胶和同轴多通道生物打印系统可以初步构建尿道空腔结构,打印后尿道细胞可在空腔结构内长期存活. 【总页数】4页(P241-244)

【作 者】皮庆猛;王晓磊;张志亮;周碧玉;任晓芸;陆毅;黄一雄;傅敏刚;范志宏

【作者单位】200129 上海市 上海交通大学医学院附属仁济医院整形外科;277300 山东省枣庄市 峄城中医院内科;200129 上海市 上海交通大学医学院附属仁济医院


整形外科;200129 上海市 上海交通大学医学院附属仁济医院整形外科;200129 上海市 上海交通大学医学院附属仁济医院整形外科;200129 上海市 上海交通大学医学院附属仁济医院整形外科;200129 上海市 上海交通大学医学院附属仁济医院整形外科;200129 上海市 上海交通大学医学院附属仁济医院整形外科;200129 上海市 上海交通大学医学院附属仁济医院整形外科 【正文语种】中 文 【中图分类】R318.1 【相关文献】

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4.不同浓度明胶水凝胶用于3D生物打印的初步研究 [J], 贺思;黄汉记;雷丹青 5.不同浓度明胶水凝胶用于3D生物打印的初步研究 [J], 贺思;黄汉记;雷丹青

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