梓醇/GelMA水凝胶的构建及其抗氧化性能的体外研究

doi:10.12439/kqhm.1005-4979.2025.03.004

摘要/Abstract

摘要：

目的：探究梓醇对大鼠施万细胞（rat Schwann cell，RSC）的作用及构建梓醇/甲基丙烯酰化明胶 (methacrylated gelatin，GelMA) 水凝胶，并在体外评价细胞生物相容性及抗氧化性能。方法：首先通过 CCK8 实验确定梓醇作用于RSC96 细胞（大鼠施万细胞系）的最佳浓度，然后通过划痕实验和 Transwell 实验分析梓醇对 RSC96 细胞迁移的影响，通过实时定量聚合酶链反应（real‐time quantitative polymerase chain reaction，RT‐qPCR）分析梓醇对细胞胶质纤维酸性蛋白（glial fibrillary acidic protein，GFAP）、神经生长因子（nerve growth factor，NGF）、脑源性神经营养因子（brain-derived neurotrophic factor，BDNF）、胶质细胞源性神经营养因子（glial cell line-derived neurotrophic factor，GDNF）mRNA 表达的影响。使用H₂O₂构建氧化应激模型，通过 CCK8 和活性氧（reactive oxygen species，ROS）染色法检测梓醇的抗氧化能力；最后检测梓醇/GelMA 水凝胶的力学性能、生物相容性、抗氧化能力及其对 RSC96 细胞增殖的作用。结果：梓醇作用于 RCS96 细胞的最佳浓度为 10 μmol/L，该浓度下梓醇能促进 RSC96 细胞的增殖和迁移，提高GFAP、NGF、BDNF、GDNF的mRNA表达，减少氧化应激对RSC96细胞的损伤。构建的梓醇/GelMA 水凝胶可以负载RSC96细胞，且具有良好的生物相容性，能促进RSC96细胞的增殖，并对处于H₂O₂环境中的细胞具有保护作用。结论：梓醇/GelMA 水凝胶具有良好的体外细胞生物相容性和抗氧化性。

关键词:

梓醇, 施万细胞, 氧化应激, 水凝胶, 体外细胞实验

Abstract:

Objective: To investigate the effects of catalpol on rat Schwann cell (RSC) and establish a catalpol/methacrylated gelatin (GelMA) hydrogel, followed by in vitro evaluation of its biocompatibility and antioxidant properties. Methods: First, the optimal concentration of catalpol for acting on RSC96 cell (rat Schwann cell line) was determined through CCK8 assay. Subsequently, the effects of catalpol on RSC96 cell migration were analyzed by using scratch assay and Transwell assay. Realtime quantitative polymerase chain reaction (RT-qPCR) was employed to assess catalpol's influence on mRNA expression levels of glial fibrillary acidic protein (GFAP), nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF). An oxidative stress model was established using H₂O₂, and catalpol's antioxidant capacity was evaluated through CCK8 assay and reactive oxygen species (ROS) staining. Finally, the mechanical properties,biocompatibility, antioxidant capacity of catalpol/GelMA hydrogel, along with its effects on RSC96 cell proliferation, were systematically investigated. Results: The optimal concentration of catalpol in GelMA for acting on RSC96 cells was determined to be 10 µmol/L. At this concentration, catalpol significantly promoted the proliferation and migration of RSC96 cell, upregulated mRNA expression levels of GFAP, NGF, BDNF and GDNF, and attenuated oxidative stress-induced damage in RSC96 cells. The constructed catalpol/GelMA hydrogel demonstrated excellent capability for RSC96 cell encapsulation and exhibited favorable biocompatibility. Furthermore, it effectively enhanced RSC96 cell proliferation and exerted protective effects against H₂O₂-induced cytotoxicity. Conclusion: Catalpol/GelMA hydrogel has good in vitro cell biocompatibility and antioxidant properties.

Key words:

catalpol, Schwann cells, oxidative stress, hydrogel, in vitro cell experiments

中图分类号:

R782

张春雷, 王迪, 杨景, 韩鸿洋, 杨澍, 逄博, 宋涛.

梓醇/GelMA水凝胶的构建及其抗氧化性能的体外研究[J]. 《口腔颌面外科杂志》, 2025, 35(3): 190-198.

ZHANG Chunlei, WANG Di, YANG Jing, HAN Hongyang, YANG Shu, PANG Bo, SONG Tao.

In vitro study on the preparation of catalpol/GelMA hydrogel and its antioxidant properties[J]. 《Journal of Oral and Maxillofacial Surgery》, 2025, 35(3): 190-198.

https://journal06.magtech.org.cn/Jweb_joms/CN/Y2025/V35/I3/190

参考文献

[1] 徐万林, 吴一凡, 杨雯君. 面神经损伤修复过程中差异表达 microRNA 的初步筛选分析 [J]. 组织工程与重建外科, 2023, 19(4): 352-357.

[2] 安玉川, 成荣, 孙蕾, 等. 一种基于光交联明胶-聚丙烯酰胺的神经组织工程构建物 [J]. 太原理工大学学报, 2023, 54(1): 170-177.

[3] Li GC, Zhang LL, Han Q, et al. Photothermal responsive cell-laden PNIPAM self-rolling hydrogel containing dopamine enhanced MWCNTs for peripheral nerve regeneration[J]. Compos Part B Eng, 2023, 254: 110551.

[4] Hsu CC, Huang HC, Wu PT, et al. Sesame oil improves functional recovery by attenuating nerve oxidative stress in a mouse model of acute peripheral nerve injury: Role of Nrf-2[J]. J Nutr Biochem, 2016, 38: 102-106.

[5] Zhu HF, Wang JH, Shao YL, et al. Catalpol may improve axonal growth via regulating miR-124 regulated PI3K/AKT/mTOR pathway in neurons after ischemia[J]. Ann Transl Med, 2019, 7(14): 306.

[6] Surmelioglu O, Sencar L, Ozdemir S, et al. Effects of agmatine sulphate on facial nerve injuries[J]. J Laryngol Otol, 2017, 131(3): 221-226.

[7] Rovak JM, Tung TH, MacKinnon SE. The surgical management of facial nerve injury[J]. Semin Plast Surg, 2004, 18(1): 23-30.

[8] Hu J, Tian LL, Prabhakaran MP, et al. Fabrication of nerve growth factor encapsulated aligned poly(ε-caprolactone) nanofibers and their assessment as a potential neural tissue engineering scaffold[J].Polymers (Basel), 2016, 8(2): 54.

[9] Li Y, Ma ZJ, Ren Y, et al. Tissue engineering strategies for peripheral nerve regeneration[J]. Front Neurol, 2021, 12: 768267.

[10] Tian YY, Jiang B, An LJ, et al. Neuroprotective effect of catalpol against MPP⁺-induced oxidative stress in mesencephalic neurons[J]. Eur J Pharmacol, 2007, 568(1-3): 142-148.

[11] Zhu HF, Shao YL, Qin L, et al. Catalpol enhances neurogenesis and inhibits apoptosis of new neurons via BDNF, but not the BDNF/trkb pathway[J]. Drug Des Devel Ther, 2019, 13: 4145-4157.

[12] He B, Hu JF, Zhu ZW. Editorial: The function of Schwann cells in peripheral nervous system[J]. Front Cell Neurosci, 2023, 16: 1129560.

[13] Kamil K, Yazid MD, Idrus RBH, et al. Hydroxytyrosol promotes proliferation of human schwann cells: An in vitro study[J]. Int J Environ Res Public Health, 2020, 17(12): 4404.

[14] Li HF, Wang YR, Huo HP, et al. Neuroprotective effects of ultrasound-guided nerve growth factor injections after sciatic nerve injury[J]. Neural Regen Res, 2015, 10(11): 1846-1855.

[15] Zuo WP, Wu HY, Zhang K, et al. Baicalin promotes the viability of Schwann cells in vitro by regulating neurotrophic factors[J]. Exp Ther Med, 2017, 14(1): 507-514.

[16] Luo XJ, Chen BG, Zheng R, et al. Hydrogen peroxide induces apoptosis through the mitochondrial pathway in rat Schwann cells[J]. Neurosci Lett, 2010, 485(1): 60-64.

[17] Bi J, Jiang B, Liu JH, et al. Protective effects of catalpol against H2O2-induced oxidative stress in astrocytes primary cultures[J]. Neurosci Lett, 2008, 442(3): 224-227.

[18] Klotz BJ, Gawlitta D, Rosenberg AJWP, et al. Gelatinmethacryloyl hydrogels: Towards biofabrication-based tissue repair[J]. Trends Biotechnol, 2016, 34(5): 394-407.

[19] Yue K, Trujillo-de Santiago G, Alvarez MM, et al. Synthesis, properties, and biomedical applications of gelatin methacryloyl (GelMA) hydrogels[J]. Biomaterials, 2015, 73: 254-271.

[20] Park J, Jeon J, Kim B, et al. Electrically conductive hydrogel nerve guidance conduits for peripheral nerve regeneration[J]. Adv Funct Mater, 2020, 30(39): 2003759.

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