Massachusetts Eye and Ear and Schepens Eye Research Institute, Department of Ophthalmology, Harvard Medical School, Boston, MA, USA.
Department of Immunology, Oslo University Hospital, Rikshospitalet, University of Oslo, Oslo, Norway.
Acta Biomater. 2019 Sep 15;96:330-344. doi: 10.1016/j.actbio.2019.07.002. Epub 2019 Jul 5.
To address the shortcomings associated with corneal transplants, substantial efforts have been focused on developing new modalities such as xenotransplantion. Xenogeneic corneas are anatomically and biomechanically similar to the human cornea, yet their applications require prior decellularization to remove the antigenic components to avoid rejection. In the context of bringing decellularized corneas into clinical use, sterilization is a crucial step that determines the success of the transplantation. Well-standardized sterilization methods, such as gamma irradiation (GI), have been applied to decellularized porcine corneas (DPC) to avoid graft-associated infections in human recipients. However, little is known about the effect of GI on decellularized corneal xenografts. Here, we evaluated the radiation effect on the ultrastructure, optical, mechanical and biological properties of DPC. Transmission electron microscopy revealed that gamma irradiated decellularized porcine cornea (G-DPC) preserved its structural integrity. Moreover, the radiation did not reduce the optical properties of the tissue. Neither DPC nor G-DPC led to further activation of complement system compared to native porcine cornea when exposed to plasma. Although, DPC were mechanically comparable to the native tissue, GI increased the mechanical strength, tissue hydrophobicity and resistance to enzymatic degradation. Despite these changes, human corneal epithelial, stromal, endothelial and hybrid neuroblastoma cells grew and differentiated on DPC and G-DPC. Thus, GI may achieve effective tissue sterilization without affecting critical properties that are essential for corneal transplant survival.
为了解决角膜移植相关的缺陷,人们投入了大量精力来开发新的方法,例如异种移植。异种角膜在解剖学和生物力学上与人类角膜相似,但它们的应用需要预先脱细胞化以去除抗原性成分,从而避免排斥反应。在将脱细胞化角膜应用于临床的过程中,灭菌是决定移植成功的关键步骤。已经应用了标准化的灭菌方法,如伽马射线照射(GI),对脱细胞化猪角膜(DPC)进行灭菌,以避免人类受者中移植物相关感染。然而,对于 GI 对脱细胞化角膜异种移植物的影响知之甚少。在这里,我们评估了 GI 对 DPC 的超微结构、光学、机械和生物学性能的影响。透射电子显微镜显示,伽马射线照射的脱细胞化猪角膜(G-DPC)保持了其结构完整性。此外,辐射并没有降低组织的光学性能。与天然猪角膜相比,DPC 和 G-DPC 在暴露于等离子体时都不会进一步激活补体系统。尽管 DPC 的机械性能与天然组织相当,但 GI 增加了机械强度、组织疏水性和抗酶降解能力。尽管存在这些变化,但人角膜上皮、基质、内皮和混合神经母细胞瘤细胞在 DPC 和 G-DPC 上生长和分化。因此,GI 可以实现有效的组织灭菌,而不会影响对角膜移植存活至关重要的关键特性。