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数字微镜器件投影打印系统用于制造半月板组织工程。

Digital micromirror device projection printing system for meniscus tissue engineering.

机构信息

Shiley Center for Orthopaedic Research and Education at Scripps Clinic, 11025 North Torrey Pines Road, Suite 200, La Jolla, CA 92037, USA.

出版信息

Acta Biomater. 2013 Jul;9(7):7218-26. doi: 10.1016/j.actbio.2013.03.020. Epub 2013 Mar 21.

Abstract

Meniscus degeneration due to age or injury can lead to osteoarthritis. Although 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 new tissue formation was assessed by gene expression analysis and histology after 2weeks in serum-free culture with transforming growth factor β1 (10ngml(-1)). Light, confocal and scanning electron microscopy were used to observe cell-GelMA interactions. Tensile mechanical testing was performed on unseeded, fresh scaffolds and 2-week-old cell-seeded and unseeded scaffolds. 2-week-old cell-GelMA constructs were implanted into surgically created meniscus defects in an explant organ culture model. No cytotoxic effects were observed 3weeks 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 the meniscus collagen bundle organization shows promise for meniscal repair. Furthermore, the technique presented may be scaled up to repair larger defects.

摘要

由于年龄或损伤导致的半月板退变可引发骨关节炎。虽然有一定的前景,但目前基于细胞的方法显示出有限的成功。在这里,我们提出了一种通过投影立体光刻技术制作的甲基丙烯酰化明胶(GelMA)支架,以模拟天然半月板组织中细胞的周向排列。将来自正常半月板的人无血管区半月板细胞接种在支架上。在无血清培养中,用转化生长因子 β1(10ngml(-1))培养 2 周后,通过基因表达分析和组织学评估监测细胞活力和新组织形成。使用明场、共聚焦和扫描电子显微镜观察细胞-GelMA 相互作用。对未接种的新鲜支架、2 周龄细胞接种和未接种支架进行拉伸力学测试。将 2 周龄的细胞-GelMA 构建体植入手术创建的半月板缺损的器官培养模型中。植入后 3 周未观察到细胞毒性作用,细胞生长并排列到图案化的 GelMA 链上。基因表达谱和组织学表明促进纤维软骨样半月板表型,并且在器官培养模型中观察到支架与修复组织的整合。我们表明,微图案化 GelMA 支架无毒,可产生有序的细胞排列,并促进半月板样组织形成。模仿半月板胶原束组织的 GelMA 支架的预制具有半月板修复的应用前景。此外,所提出的技术可以扩展到修复更大的缺陷。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3bb2/3685281/c92a933de554/nihms462241f1.jpg

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