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Reconstruction of ocular surfaces with limbal stem cells supported by porous composite hydrogels

多孔复合水凝胶负载角膜缘干细胞重建眼表

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Abstract

Stem cell-based therapy is a novel technique used to reconstruct the ocular surface of patients with limbal stem cell deficiency (LSCD). However, the reported stem cell therapies for LSCD are limited to two-dimensional (2D) culture systems, which cannot mimic the native microenvironment in which stem cells reside in vivo. Herein, an injectable photocrosslinkable hydrogel was developed based on porous gelatin methacryloyl (GelMA)/silk fibroin glycidyl methacrylate (SilMA), which encapsulated limbal stem cells (LSCs) to reconstruct ocular surfaces. The porous GelMA/SilMA hydrogels demonstrated excellent transparency and good adhesion. More importantly, this hydrogel maintained viability, supported the adhesion of encapsulated rabbit LSCs and promoted their migration in vitro in three-dimensional (3D) culture environments. Proteomic analysis confirmed that the porous GelMA/SilMA hydrogel 3D cell culture system maintained both the self-renewal and differentiation of the LSCs. Furthermore, histological analyses demonstrated that the porous GelMA/SilMA hydrogel encapsulating LSCs effectively promoted the re-epithelization of LSCD. Porous hydrogels were subjected to simplified serial cell culture operations to construct an ocular surface with a uniform cell distribution. These findings provide valuable insights into stem cell therapies and regenerative medicine for ocular surface reconstruction.

摘要

干细胞疗法是一种用于重建角膜缘干细胞缺乏症(LSCD)患者眼 表的新技术. 然而, 目前报道的干细胞疗法治疗LSCD仅局限于二维 (2D)培养体系, 不能模拟干细胞在体内生存的原生微环境. 本研究基于 多孔明胶甲基丙烯酰(GelMA)/丝素蛋白甲基丙烯酸缩水甘油酯(SilMA), 制备了一种可注射的光交联水凝胶用于负载角膜缘干细胞 (LSCs)以重建眼表. 多孔GelMA/SilMA水凝胶具有优异的透明度和良 好的粘附性. 更重要的是, 该水凝胶保持了兔LSCs的活力, 支持了兔 LSCs的粘附, 并促进了它们在体外3D培养环境中的迁移. 蛋白质组学 分析证实, 多孔GelMA/SilMA水凝胶3D细胞培养体系能够保持LSCs的 自我更新和分化能力. 此外, 组织学分析表明, 负载LSCs的多孔GelMA/SilMA水凝胶能够有效的促进LSCD再上皮化. 多孔水凝胶能够简化连 续细胞培养的相关操作, 以构建具有细胞均匀分布的眼表. 这些发现为 干细胞疗法和再生医学用于眼表重建提供了有价值的见解.

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Acknowledgements

This work was funded by the National Natural Science Foundation of China (82201153), the Natural Science Foundation of Shandong Province (ZR2022QH276), the National Key Research and Development Program of China (2023YFE0206700), the Key Research and Development Program of Shandong Province (2021ZDSYS14), the Academic Promotion Program of Shandong First Medical University (2019ZL001 and 2019RC008), and the Taishan Scholar Program (20161059).

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Authors and Affiliations

  1. Eye Institute of Shandong First Medical University, State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, School of Ophthalmology, Shandong First Medical University, Qingdao, 266071, China

    Fuyan Wang  (王付燕), Yuehe Xu  (许悦荷), Depeng Shi  (石德鹏), Jun Cheng  (程钧), Long Zhao  (赵龙), **a Qi  (祁霞), Qingjun Zhou  (周庆军) & Lixin **e  (谢立信)

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  1. Fuyan Wang  (王付燕)
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  2. Yuehe Xu  (许悦荷)
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  3. Depeng Shi  (石德鹏)
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  4. Jun Cheng  (程钧)
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  5. Long Zhao  (赵龙)
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  6. **a Qi  (祁霞)
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  7. Qingjun Zhou  (周庆军)
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  8. Lixin **e  (谢立信)
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Contributions

Author contributions Wang F, Zhou Q and **e L conceived and designed the experiments. Wang F, Xu Y, Shi D, Cheng J, Zhao L and Qi X carried out the experiments. Wang F and Xu Y analyzed the data. Wang F, Zhou Q and **e L drafted and revised the paper. All authors participated in data discussionst. All authors read and approved the publication of the final manuscript.

Corresponding authors

Correspondence to Qingjun Zhou  (周庆军) or Lixin **e  (谢立信).

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Conflict of interest The authors declare that they have no conflict of interest.

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Fuyan Wang received her MD degree from Shandong University in 2021 and then worked as an ophthalmologist at Eye Institute of Shandong First Medical University. Her interests include the basic and clinical research of tissue-engineered cornea and infections corneal diseases.

Qingjun Zhou received his PhD degree from Zhejiang University in 2006. He now serves as the executive deputy director of Shandong Provincial Key Laboratory of Ophthalmology. His interests include the basic and clinical research of corneal injury repair and functional reconstruction.

Lixin **e received his MD degree from Shandong Medical College (now Shandong University) in 1965. Since 1991, he established and worked as the director of Shandong Eye Institute. His interests include the basic and clinical research of infections corneal diseases, diabetic ocular diseases, tissue-engineered cornea and cataract.

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Wang, F., Xu, Y., Shi, D. et al. Reconstruction of ocular surfaces with limbal stem cells supported by porous composite hydrogels. Sci. China Mater. (2024). https://doi.org/10.1007/s40843-024-2970-2

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