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Digital-Micromirror-Device Projection Printing System for Meniscus Tissue Engineering

机译:MENSISCUS组织工程的数码微镜 - 器件投影印刷系统

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摘要

Meniscus degeneration due to age or injury can lead to osteoarthritis. Though promising, current cell-based approaches show limited success. Here we present three-dimensional methacrylated gelatin (GelMA) scaffolds patterned via projection stereolithography to emulate the circumferential alignment of cells in native meniscus tissue. Cultured human avascular zone meniscus cells from normal meniscus were seeded on the scaffolds. Cell viability was monitored, and neo-tissue formation was assessed by gene expression analysis and histology after two weeks in serum free culture with TGFβ1 (10ng/ml). Light, confocal and scanning electron microscopy was used to observe cell/GelMA interactions. Tensile mechanical testing was performed on unseeded, fresh scaffolds and two-week old cell-seeded and unseeded scaffolds. Two-week old cell/GelMA constructs were implanted into surgically created meniscus defects in an explant organ culture model. No cytotoxic effects were observed three weeks after implantation, and cells grew and aligned to the patterned GelMA strands. Gene expression profiles and histology indicated promotion of a fibrocartilage-like meniscus phenotype, and scaffold integration with repair tissue was observed in the explant model. We show that micropatterned GelMA scaffolds are non-toxic, produce organized cellular alignment, and promote meniscus-like tissue formation. Prefabrication of GelMA scaffolds with architectures mimicking meniscus collagen bundle organization shows promise for meniscal repair. Furthermore, the technique presented may be scaled to repair larger defects.
机译:由于年龄或受伤造成的半月板变性会导致骨关节炎。尽管很有希望,但是当前基于单元的方法显示出有限的成功。在这里,我们介绍通过投影立体光刻图案化的三维甲基丙烯酸明胶(GelMA)支架,以模拟天然半月板组织中细胞的周向排列。将来自正常弯液面的培养的人无血管区弯液面细胞接种在支架上。监测细胞存活力,并在无血清的TGFβ1(10ng / ml)培养中两周后通过基因表达分析和组织学评估新组织的形成。使用光,共聚焦和扫描电子显微镜观察细胞/ GelMA相互作用。对未播种的新鲜支架和两周龄的细胞播种和未播种的支架进行拉伸机械测试。将两周大的细胞/ GelMA构建体植入到外植体器官培养模型中由手术产生的半月板缺陷中。植入三周后未观察到细胞毒性作用,并且细胞生长并与图案化的GelMA链对齐。基因表达谱和组织学表明促进了纤维软骨样半月板表型,并在外植体模型中观察到支架与修复组织的整合。我们表明,micropatterned的GelMA支架是无毒的,产生有组织的细胞排列,并促进半月板样组织形成。模仿半月板胶原束组织结构的GelMA支架的预制显示了半月板修复的前景。此外,提出的技术可以按比例缩放以修复较大的缺陷。

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