Department of Ophthalmology, Clinical Medical College of Shandong University, China; State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, China.
State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, China; Eye Hospital of Shandong First Medical University, China.
Ocul Surf. 2020 Oct;18(4):748-760. doi: 10.1016/j.jtos.2020.07.020. Epub 2020 Aug 22.
Hydrogels derived from decellularized tissues provide superior biocompatibility, tenability and tissue-specific extracellular matrix (ECM) components. Based on the preparation of decellularized porcine cornea (DPC), here we developed an injectable and transparent hydrogel for the regeneration of epithelium and stroma in focal corneal defects.
The DPC-derived hydrogel was prepared with N-cyclohexyl-N'-(2-morpholinethyl) carbodiimide metho-p-toluenesulfonate/N-hydroxysuccinimide (CMC/NHS) as cross-linkers. The characteristics of the hydrogel were analyzed and its cytocompatibility was assessed by Live/Dead and Cell Counting Kit (CCK)-8 assays. Immunofluorescence staining, quantitative PCR and Western blot analyses were performed to assess the relative protein and gene expression in corneal fibroblasts on hydrogel. The safety and efficiency of the hydrogel for repairing focal corneal defects in rabbit were measured by slit-lamp, anterior segment optical coherence tomography (AS-OCT), confocal microscopy and histological analyses.
The DPC-derived hydrogel cross-linked with CMC/NHS assumed favorable transparency, exhibited distinct mechanical properties and preserved the ECM components of native porcine cornea (NPC). In vitro experiments showed that the hydrogel maintained the phenotype, supported the proliferation and promoted the ECM synthesis of corneal fibroblasts. When injected onto rabbit corneas, the hydrogel rapidly covered, solidified and formed a smooth surface on the focal defect. Corneal epithelium was fully regenerated within 3 days. The thickness of the corneal epithelium and stroma was restored at 12 weeks after surgery without significant inflammation or scar formation. Notably, the hydrogel showed no harmful effects on the resident stroma and endothelium.
The DPC-derived hydrogel may represent a promising biomaterial for corneal epithelial and stromal regeneration.
脱细胞组织衍生的水凝胶提供了卓越的生物相容性、可调节性和组织特异性细胞外基质（ECM）成分。基于脱细胞猪角膜（DPC）的制备，我们开发了一种可注射的透明水凝胶，用于再生局灶性角膜缺损的上皮和基质。
使用 N-环己基-N'-(2-吗啉代乙基)碳二亚胺甲氧基-对甲苯磺酸盐/N-羟基琥珀酰亚胺（CMC/NHS）作为交联剂制备 DPC 衍生的水凝胶。分析水凝胶的特性，并通过 Live/Dead 和细胞计数试剂盒（CCK-8）测定评估其细胞相容性。通过免疫荧光染色、定量 PCR 和 Western blot 分析评估角膜成纤维细胞在水凝胶上的相对蛋白和基因表达。通过裂隙灯、眼前节光学相干断层扫描（AS-OCT）、共聚焦显微镜和组织学分析测量水凝胶修复兔局灶性角膜缺损的安全性和效率。
用 CMC/NHS 交联的 DPC 衍生水凝胶具有良好的透明度，表现出明显的机械性能，并保留了天然猪角膜（NPC）的 ECM 成分。体外实验表明，水凝胶维持了表型，支持增殖并促进了角膜成纤维细胞的 ECM 合成。当注入兔角膜时，水凝胶迅速覆盖、凝固并在局灶性缺损处形成光滑表面。角膜上皮在 3 天内完全再生。术后 12 周，角膜上皮和基质的厚度得到恢复，没有明显的炎症或瘢痕形成。值得注意的是，水凝胶对固有基质和内皮没有不良影响。
DPC 衍生的水凝胶可能代表一种有前途的用于角膜上皮和基质再生的生物材料。
