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包裹牙髓干细胞的注射性水凝胶用于治疗外伤性视神经损伤。

Injective hydrogel encapsulating dental pulp stem cells for the treatment of traumatic optic nerve injury.

作者信息

Luo Lihua, Xing Zhenjie, Li Yao, Wang Ben, Dong Na, Sun Jiayi, Wang Shuimiao, Xu Yidi, He Yan, Li Lingli, Nan Kaihui, Ye Qingsong

机构信息

School and Hospital of Stomatology, Wenzhou Medical University, Wenzhou, Zhejiang, China.

The Eye Hospital, School of Ophthalmology and Optometry, Wenzhou Medical University, Wenzhou, Zhejiang, China.

出版信息

Front Bioeng Biotechnol. 2025 Feb 25;13:1528749. doi: 10.3389/fbioe.2025.1528749. eCollection 2025.

Abstract

OBJECTIVES

The study aimed to evaluate the effect of GeLMA/bFGF hydrogel loaded with dental pulp stem cells (DPSCs) on the repair and regeneration of traumatic optic nerve injury.

MATERIALS AND METHODS

GeLMA/bFGF hydrogel was photo-cross-linked by LED light. The physical-chemical properties and cytocompatibility of GeLMA/bFGF hydrogel after being squeezed (GeLMA/bFGF-SQ) were evaluated by SEM and degradation analyses, as well as live/dead and CCK-8 assays, respectively. The axon growth of PC12 cells was evaluated by MAP2 staining. The GeLMA/bFGF/DPSCs-SQ hydrogel was injected into the lesion site to observe the repair of optic nerve injury. The number of retinal ganglion cells (RGCs) was calculated by βIII-tubulin staining. The length of regenerated axons was observed by CTB staining. Tissue recovery and axon growth of the optic nerve were observed by HE and TEM analyses, respectively.

RESULTS

GeLMA/bFGF-SQ hydrogel had a porous structure and great degradability, as well as good cytocompatibility. Meanwhile, DPSCs-conditioned medium (DPSCs-CM) could promote the axon growth of PC12 cells. Moreover, the number of RGCs and the regeneration of axons of the optic nerve were the highest in the GeLMA/bFGF/DPSCs-SQ group. HE and TEM data revealed abundant newly and orderly arrangement of optic nerve axons that was observed in the damaged area of the GeLMA/bFGF/DPSCs-SQ group.

CONCLUSION

Transplantation of GeLMA/bFGF/DPSCs-SQ provided an appropriate microenvironment for the repair and regeneration of injured optic nerves. Moreover, DPSCs combined with bFGF protected the RGCs from apoptosis and promoted optic nerve regeneration by secreting a series of neurotrophic factors.

摘要

目的

本研究旨在评估负载牙髓干细胞(DPSCs)的GeLMA/bFGF水凝胶对创伤性视神经损伤修复和再生的影响。

材料与方法

GeLMA/bFGF水凝胶通过LED光进行光交联。分别通过扫描电子显微镜(SEM)和降解分析以及活/死细胞检测和CCK-8检测评估挤压后的GeLMA/bFGF水凝胶(GeLMA/bFGF-SQ)的物理化学性质和细胞相容性。通过微管相关蛋白2(MAP2)染色评估PC12细胞的轴突生长。将GeLMA/bFGF/DPSCs-SQ水凝胶注入损伤部位以观察视神经损伤的修复情况。通过βIII微管蛋白染色计算视网膜神经节细胞(RGCs)的数量。通过霍乱毒素B亚基(CTB)染色观察再生轴突的长度。分别通过苏木精-伊红(HE)染色和透射电子显微镜(TEM)分析观察视神经的组织恢复和轴突生长情况。

结果

GeLMA/bFGF-SQ水凝胶具有多孔结构和良好的降解性,以及良好的细胞相容性。同时,DPSCs条件培养基(DPSCs-CM)可促进PC12细胞的轴突生长。此外,GeLMA/bFGF/DPSCs-SQ组的RGCs数量和视神经轴突再生情况最佳。HE和TEM数据显示,在GeLMA/bFGF/DPSCs-SQ组的损伤区域观察到大量新的且有序排列的视神经轴突。

结论

GeLMA/bFGF/DPSCs-SQ移植为受损视神经的修复和再生提供了适宜的微环境。此外,DPSCs与碱性成纤维细胞生长因子(bFGF)联合通过分泌一系列神经营养因子保护RGCs免于凋亡并促进视神经再生。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc48/11893850/a91af312fad5/fbioe-13-1528749-g001.jpg

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