Department of Ophthalmology, Clinical Medical College of Shandong University, Jinan, China.
State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China.
Xenotransplantation. 2020 Nov;27(6):e12633. doi: 10.1111/xen.12633. Epub 2020 Jul 29.
The sclera is one of the most commonly used repair materials in ophthalmic plastic surgery and is often used for supporting, wrapping, filling, and pressing during surgery. Although the sclera plays an irreplaceable role in ophthalmology applications, there are many restrictive factors, such as high costs and limited sources. Here, we report the use of a decellularized porcine sclera (DPS) for scleral reconstruction in rabbit models.
The DPS generated by a hybrid decellularization protocol was characterized in respect of histological observation, DNA, α-gal, GAG, and collagen content. The mechanical properties were evaluated by uniaxial tensile testing. LIVE/DEAD and Cell Counting Kit (CCK)-8 assays were performed to assess its in vitro cytocompatibility and cytotoxicity. In vivo biocompatibility and biointegration of the DPS for repairing scleral defect in rabbit were measured by slit-lamp and histological analyses. Immunohistochemical (IHC) staining was used to detect the expression of CD4, CD8, CD45, CD68, and vimentin.
Through decellularization, the major xenoantigen DNA and α-gal are efficiently removed while abundant matrix components and mechanical properties are well preserved in the DPS. Extracts of the DPS and DPS samples had no inhibitory effects on the proliferation of HFSFs. Moreover, there was no sign that an immune reaction occurred in or around the transplanted DPS grafts within 28 days of animal implantation.
The decellularization strategy we developed is feasible and effective. The prepared DPS holds great potential for the repair of scleral injury.
巩膜是眼科整形手术中最常用的修复材料之一，常用于手术中的支撑、包裹、填充和压迫。尽管巩膜在眼科应用中具有不可替代的作用，但存在许多限制因素，例如成本高和来源有限。在这里，我们报告了使用脱细胞猪巩膜（DPS）进行兔模型巩膜重建。
通过混合脱细胞方案生成的 DPS 进行了组织学观察、DNA、α-半乳糖、GAG 和胶原蛋白含量的特征描述。通过单轴拉伸试验评估了机械性能。通过 LIVE/DEAD 和细胞计数试剂盒（CCK-8）测定评估了其体外细胞相容性和细胞毒性。通过裂隙灯和组织学分析测量 DPS 修复兔巩膜缺损的体内生物相容性和生物整合。免疫组织化学（IHC）染色用于检测 CD4、CD8、CD45、CD68 和波形蛋白的表达。
通过脱细胞处理，有效地去除了主要的异种抗原 DNA 和α-半乳糖，同时在 DPS 中保留了丰富的基质成分和良好的机械性能。DPS 的提取物和 DPS 样本对 HFSFs 的增殖没有抑制作用。此外，在动物植入后 28 天内，移植的 DPS 移植物内或周围没有发生免疫反应的迹象。
我们开发的脱细胞策略是可行且有效的。所制备的 DPS 具有修复巩膜损伤的巨大潜力。
