Conjunctival defects severely impair ocular surface homeostasis and vision. However, the existing conjunctival reconstruction methods are still unable to achieve satisfactory functional replacement of conjunctiva. Scaffold materials that provide a more stem-cell-friendly microenvironment are in urgent need of improvement. Herein we reported a functional conjunctival reconstruction scaffold constructed from decellularized matrix derived from rabbit subconjunctival fibroblasts (DM-SCF) loaded with conjunctival epithelial stem cells (CjESCs) for ocular surface repair. The DM-SCF had a thickness and mechanical strength close to that of normal conjunctiva and showed excellent biocompatibility. CjESCs inoculated on DM-SCF maintained good stem cell properties, which showed long-term maintenance of low differentiation and good proliferation activity. DM-SCF transplantation with CjESCs showed satisfactory results in rabbit models of a large conjunctival defect. The conjunctiva was reconstructed with abundant goblet cell expression and regular fiber arrangement. Protein mass spectrometry revealed the active extracellular matrix components of DM-SCF, regulating the activation of Wnt/β-catenin and Notch pathways for stem cell maintenance. Overall, our study developed a promising new strategy for ocular surface reconstruction by optimizing bioactive scaffold and providing functional stem cell niche, so as to achieve long-standing and superior functional repair.

结膜缺陷严重损害眼表稳态和视力。 然而，现有的结膜重建方法仍无法实现满意的结膜功能置换。提供对干细胞更友好的微环境的支架材料迫切需要改进。在这里，我们报道了一种功能性结膜重建支架，该支架由脱细胞基质构成，该基质来源于兔结膜下成纤维细胞 (DM-SCF)，负载结膜上皮干细胞 (CjESCs)，用于眼表修复。DM-SCF具有接近正常结膜的厚度和机械强度，并表现出优异的生物相容性。接种在DM-SCF上的CjESCs保持良好的干细胞特性，长期维持低分化和良好的增殖活性。 CjESCs 的 DM-SCF 移植在大结膜缺损的兔模型中显示出令人满意的结果。 结膜重建，杯状细胞表达丰富，纤维排列规则。蛋白质质谱分析揭示了 DM-SCF 的活性细胞外基质成分，调节 Wnt/β-catenin 和 Notch 通路的激活以维持干细胞。总体而言，我们的研究通过优化生物活性支架和提供功能性干细胞生态位，开发了一种有前景的眼表重建新策略，从而实现长期和卓越的功能修复。