中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (30): 7880-7892.doi: 10.12307/2026.964

• 神经组织构建 nerve tissue construction • 上一篇    下一篇

神经生长因子-壳聚糖水凝胶对视网膜外植体的神经保护和促再生作用

马新月，段红梅，郝  鹏，赵  文，高钰丹，郭旬会，姜蕙汀，许玲瑞，杨朝阳，李晓光   

  1. 首都医科大学基础医学院，北京市   100069
  • 收稿日期:2026-04-11 修回日期:2026-06-26 出版日期:2026-10-28 发布日期:2026-09-11
  • 通讯作者: 李晓光，博士，教授，首都医科大学基础医学院，北京市 100069 通讯作者：杨朝阳，博士，教授，首都医科大学基础医学院，北京市 100069
  • 作者简介:马新月，女，1999年生，河北省承德市人，满族，硕士，主要从事生物活性材料治疗视网膜神经退行性疾病方面的研究。
  • 基金资助:
    国家自然科学基金项目(82271403，81941011，31730030)，项目负责人：李晓光；国家自然科学基金项目(82272171，31971279)，项目负责人：杨朝阳；北京市自然科学基金项目(7222004)，项目负责人：段红梅

Nerve growth factor-chitosan hydrogels: neuroprotective and pro-regenerative effects on retinal explants

Ma Xinyue, Duan Hongmei, Hao Peng, Zhao Wen, Gao Yudan, Guo Xunhui, Jiang Huiting, Xu Lingrui, Yang Chaoyang, Li Xiaoguang   

  1. School of Basic Medicine, Capital Medical University, Beijing 100069, China
  • Received:2026-04-11 Revised:2026-06-26 Online:2026-10-28 Published:2026-09-11
  • Contact: Yang Chaoyang, PhD, Professor, School of Basic Medicine, Capital Medical University, Beijing 100069, China
  • About author:Ma Xinyue, MS, School of Basic Medicine, Capital Medical University, Beijing 100069, China Corresponding author: Li Xiaoguang, PhD, Professor, School of Basic Medicine, Capital Medical University, Beijing 100069, China
  • Supported by:
    National Natural Science Foundation of China, Nos. 82271403, 81941011, 31730030 (to LXG), 82272171, 31971279 (to YCY); Beijing Natural Science Foundation, No. 7222004 (to DHM)

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摘要：



文题释义：
视网膜神经节细胞：是位于视网膜最内层的唯一输出神经元，负责将光信号经视神经传递至大脑视觉中枢，是视觉形成的关键环节。其树突接收双极细胞信号，轴突汇聚形成视神经，结构完整性直接决定视觉传导功能。在青光眼、视网膜色素变性等疾病中，神经节细胞会发生不可逆凋亡，导致视力丧失甚至失明。
视网膜外植体：是从实验动物眼球中完整剥离、保留视网膜天然组织结构的体外器官型培养模型，能高度模拟在体视网膜的细胞构成、层状排列与神经环路联系，是研究视网膜神经退行性疾病的理想平台。它保留了神经节细胞、双极细胞、小胶质细胞等多种细胞类型及突触连接，可长期维持组织形态与生理特征，避免体内实验干扰因素多、操作复杂、动物消耗大等问题。

背景：视网膜神经退行性疾病以视网膜神经元进行性丢失、神经纤维及突触结构损伤为核心病理改变，目前临床治疗手段有限，亟需开发有效的神经保护与再生策略。神经生长因子和壳聚糖在神经元存活、分化及突触可塑性调控中发挥关键作用。
目的：通过构建神经生长因子-壳聚糖水凝胶缓释体系，明确其对视网膜外植体的神经保护与促再生作用。
方法：以Wistar乳鼠视网膜外植体、原代视网膜神经节细胞及视网膜神经干细胞为研究模型，采用自制神经生长因子-壳聚糖水凝胶进行干预；运用苏木精-伊红染色、免疫荧光染色、TUNEL凋亡检测、RT-qPCR及细胞培养等技术方法，系统检测视网膜组织结构、神经节细胞与双极细胞存活、细胞凋亡水平、神经突再生、神经干细胞分化以及小胶质细胞活化与极化状态等指标。
结果与结论：①神经生长因子-壳聚糖水凝胶可显著降低视网膜外植体细胞凋亡率、维持视网膜结构完整性以及缓解视网膜神经细胞进行性丢失；②神经生长因子-壳聚糖水凝胶可显著促进视网膜神经突生长且激活视网膜内源性神经干细胞，促进其增殖并定向分化为神经节细胞；③同时还发现神经生长因子-壳聚糖水凝胶可有效缓解小胶质细胞活化并且调节炎症因子表达，促进其从促炎的M1型向抗炎的M2型极化，从而构建适宜的神经再生微环境；④证实神经生长因子-壳聚糖水凝胶可通过调控小胶质细胞表型极化、改善炎症微环境，达到保护视网膜外植体神经细胞的存活和促神经发生效应，为体内应用神经生长因子-壳聚糖水凝胶治疗视网膜神经退行性疾病提供新策略。

https://orcid.org/0000-0001-7112-9854 (马新月) 


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 视网膜外植体, 神经生长因子-壳聚糖水凝胶, 小胶质细胞极化, 神经保护, 神经发生

Abstract: BACKGROUND: Retinal neurodegenerative diseases are characterized by progressive loss of retinal neurons and damage to nerve fibers and synaptic structures as the core pathological changes. Current clinical treatment options are limited, and there is an urgent need to develop effective neuroprotective and regenerative strategies. Nerve growth factor and chitosan play key roles in regulating neuronal survival, differentiation, and synaptic plasticity.
OBJECTIVE: To construct a nerve growth factor-chitosan hydrogel sustained-release system and clarify its neuroprotective and pro-regenerative effects on retinal explants.
METHODS: Retinal explants, primary retinal ganglion cells, and retinal neural stem cells from Wistar neonatal rats were used as research models and intervened with self-made nerve growth factor-chitosan hydrogels. Hematoxylin-eosin staining, immunofluorescence staining, TUNEL apoptosis assay, RT-qPCR, and cell culture techniques were employed to systematically detect retinal tissue structure, survival of ganglion cells and bipolar cells, apoptosis level, neurite regeneration, neural stem cell differentiation, and microglial activation and polarization status.
RESULTS AND CONCLUSION: (1) The nerve growth factor-chitosan hydrogel significantly reduced the apoptosis rate of retinal explants, maintained retinal structural integrity, and alleviated the progressive loss of retinal neurons. (2) The nerve growth factor-chitosan hydrogel significantly promoted retinal neurite outgrowth, activated endogenous neural stem cells in the retina, and promoted their proliferation and directional differentiation into ganglion cells. (3) It was also found that the nerve growth factor-chitosan hydrogel effectively alleviated microglial activation, regulated the expression of inflammatory factors, and promoted their polarization from the pro-inflammatory M1 phenotype to the anti-inflammatory M2 phenotype, thereby constructing a favorable microenvironment for nerve regeneration. Overall, these findings confirm that the nerve growth factor-chitosan hydrogel protects the survival of retinal explant neurons and promotes neurogenesis by regulating microglial phenotypic polarization and improving the inflammatory microenvironment, providing a new strategy for the in vivo application of nerve growth factor-chitosan hydrogel in the treatment of retinal neurodegenerative diseases.

Key words: retinal explant, nerve growth factor-chitosan hydrogel, microglial polarization, neuroprotection, neurogenesis

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