Neonates with congenital diaphragmatic hernia often require surgical defect closure with a patch. Alternatives to native diaphragmatic tissue are critically needed for this paediatric surgery. The clinical efficacy of mesh patches is limited by complications associated with residual foreign material and by hernia recurrence. In this study, we used a novel bio-3D printer method to generate large scaffold-free tissue patches composed of human cells. The resulting large tissue constructs had high elasticity and strength. Cellular patches were transplanted into rats with surgically created diaphragmatic defects. Rats survived for over 710 days after implantation of tissue constructs. CT confirmed complete tissue integration of the grafts during rat growth. Histology revealed regeneration of muscle structure, neovascularization, and neuronal networks within the reconstructed diaphragms. Our results demonstrate that created cellular patches are a highly safe and effective therapeutic strategy for repairing diaphragmatic defects, and thus pave the way for a clinical trial.

先天性diaphragm肌疝的新生儿通常需要用补丁修补手术缺损。对于这种小儿外科手术，迫切需要替代天然diaphragm肌组织。网片的临床疗效受到与残留异物相关的并发症和疝复发的限制。在这项研究中，我们使用了一种新型的bio-3D打印机方法来生成由人细胞组成的大型无支架组织斑块。所得的大组织构造物具有高弹性和强度。将细胞贴剂移植到具有手术引起的diaphragm肌缺损的大鼠中。植入组织构建物后，大鼠存活了710天以上。CT证实了大鼠生长过程中移植物的完整组织整合。组织学显示肌肉结构再生，新血管形成，和重建隔膜内的神经元网络。我们的结果表明，产生的细胞膜片是修复diaphragm肌缺损的高度安全有效的治疗策略，因此为临床试验铺平了道路。