Wang Leying, Nie Xiongfeng, Wei Yuan, Chen Qiankun, Sun Yage, Zhao Xinrui, Xu Xizhan, Liu Wenguang, Liang Qingfeng
Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing 100005, China.
School of Material Science and Engineering, Tianjin Key Laboratory of Composite and Functional Materials, Tianjin University, Tianjin 300350, China.
Acta Biomater. 2025 Jan 24;193:157-170. doi: 10.1016/j.actbio.2025.01.011. Epub 2025 Jan 9.
Limbal stem cell deficiency (LSCD) causes vision loss and is often treated by simple corneal epithelial cell transplantation with poor long-term efficiency. Here, we present a biomimetic bilayer limbal implant using digital light processing 3D printing technology with gelatin methacrylate (GelMA) and poly (ethylene glycol) diacrylate (PEGDA) bioinks containing corneal epithelial cells (CECs) and corneal stromal stem cells (CSSCs), which can transplant CECs and improve the limbal niche simultaneously. The GelMA/PEGDA hydrogel possessed robust mechanical properties to support surgical transplantation and had good transparency, suitable swelling and degradation rate as a corneal implant. Encapsulated CECs and CSSCs maintained viability and proliferative activity in the bilayer limbal implant. In vivo, both CEC-loaded and CEC/CSSC-loaded hydrogel could repair the corneal surface in the LSCD model effectively. Notably, the corneal epithelial healing was faster, and corneal opacity and neovascularization were minimal in CEC/CSSC-loaded group. These findings highlight the feasibility of 3D printing in limbal construction, providing CEC/CSSC-loaded limbal implants as a treatment strategy for LSCD and corneal blindness. STATEMENT OF SIGNIFICANCE: This study aimed to enhance the long-term prognosis of limbal epithelial cell transplantation in patients with limbal stem cell deficiency by developing a 3D limbal implant that encapsulates corneal epithelial cells and limbal niche cells simultaneously. The 3D printed implant offers the advantages of mimicking the natural layered limbal structure and were found to enhance the regenerative capacity of corneal epithelial cells, suppress inflammation, and alleviate corneal scarring in vivo. This study highlights the importance of limbal microenvironment remodeling in the treatment of limbal stem cell deficiency and the potential of 3D printing in the treatment of corneal diseases.
角膜缘干细胞缺乏症（LSCD）会导致视力丧失，目前常通过简单的角膜上皮细胞移植进行治疗，但长期疗效不佳。在此，我们展示了一种采用数字光处理3D打印技术制备的仿生双层角膜缘植入物，其使用了含有角膜上皮细胞（CECs）和角膜基质干细胞（CSSCs）的甲基丙烯酸明胶（GelMA）和聚乙二醇二丙烯酸酯（PEGDA）生物墨水，该植入物能够同时移植CECs并改善角膜缘微环境。GelMA/PEGDA水凝胶具有强大的机械性能以支持手术移植，并且作为角膜植入物具有良好的透明度、合适的溶胀率和降解率。封装在双层角膜缘植入物中的CECs和CSSCs保持了活力和增殖活性。在体内，负载CECs和负载CEC/CSSC的水凝胶均可有效修复LSCD模型中的角膜表面。值得注意的是，在负载CEC/CSSC的组中，角膜上皮愈合更快，角膜混浊和新生血管形成最少。这些发现突出了3D打印在角膜缘构建中的可行性，为负载CEC/CSSC的角膜缘植入物作为LSCD和角膜盲的治疗策略提供了依据。重要性声明：本研究旨在通过开发一种同时封装角膜上皮细胞和角膜缘微环境细胞的3D角膜缘植入物，提高角膜缘干细胞缺乏症患者角膜缘上皮细胞移植的长期预后。3D打印的植入物具有模仿天然分层角膜缘结构的优势，并且在体内被发现可增强角膜上皮细胞的再生能力、抑制炎症并减轻角膜瘢痕形成。本研究突出了角膜缘微环境重塑在角膜缘干细胞缺乏症治疗中的重要性以及3D打印在角膜疾病治疗中的潜力。
