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仿生聚(γ-苄基-L-谷氨酸)纤维支架促进角膜神经再生的研究。

Exploration of biomimetic poly(γ-benzyl-L-glutamate) fibrous scaffolds for corneal nerve regeneration.

机构信息

Department of Materials Science and Engineering, National Taiwan University, Taipei, Taiwan.

Department of Ophthalmology, National Taiwan University College of Medicine, Taipei, Taiwan.

出版信息

J Mater Chem B. 2022 Aug 24;10(33):6372-6379. doi: 10.1039/d2tb01250b.

Abstract

Poly(γ-benzyl-L-glutamate) (PBG) made biomimetic scaffolds are explored as candidate materials for corneal nerve regeneration and neurotrophic keratopathy treatment. The PBG with built-in neurotransmitter glutamate was synthesized and fabricated into 3D fibrous scaffolds containing aligned fibers using electrospinning. In experiments, primary mouse trigeminal ganglia (TG) cells were used. Immunohistochemistry (IHC) analysis shows that TG cells cultured on PBG have no cytotoxic response for 21 days. Without any nerve growth factor, TG cells have the longest neurite length of 225.3 μm in the PBG group and 1.3 times the average length as compared with the polycaprolactone and no scaffold groups. Also, aligned fibers guide the neurite growth and extension unidirectionally. assays were carried out by intracorneal implantation of PBG on clinical New Zealand rabbits. The external eye photos and confocal microscopy (IVCM) show a low immune response. The corneal neural markers (βIII tubulin and SMI312) in the IHC analysis are consistent with the position stained by glutamate of implanted scaffolds, indicating that PBG induces neurogenesis. PBG exhibits mechanical stiffness to resist material deformation possibly caused by surgical operations. The results of this study demonstrate that PBG is suitable for corneal nerve regeneration and the treatment of neurotrophic keratopathy.

摘要

聚(γ-苄基-L-谷氨酸酯)(PBG)仿生支架被探索作为角膜神经再生和神经营养性角膜病变治疗的候选材料。合成了内置神经递质谷氨酸的 PBG,并使用静电纺丝将其制成含有取向纤维的 3D 纤维支架。在实验中,使用原代小鼠三叉神经节(TG)细胞。免疫组织化学(IHC)分析表明,在 PBG 上培养的 TG 细胞在 21 天内没有细胞毒性反应。在没有任何神经生长因子的情况下,TG 细胞在 PBG 组中的最长神经突长度为 225.3 μm,是聚己内酯和无支架组的平均长度的 1.3 倍。此外,取向纤维可单向引导神经突生长和延伸。通过在临床新西兰兔的角膜内植入 PBG 进行了测定。外部眼部照片和共聚焦显微镜(IVCM)显示出低免疫反应。免疫组织化学分析中的角膜神经标志物(βIII 微管蛋白和 SMI312)与植入支架中谷氨酸染色的位置一致,表明 PBG 诱导了神经发生。PBG 具有机械刚度,可抵抗手术操作可能导致的材料变形。这项研究的结果表明,PBG 适合用于角膜神经再生和神经营养性角膜病变的治疗。

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