Network of Excellence for Functional Biomaterials (NFB), ‡Department of Anatomy, National University of Ireland , Galway, Ireland.
ACS Chem Neurosci. 2013 Sep 18;4(9):1297-304. doi: 10.1021/cn400080h. Epub 2013 Jul 8.
Neurotrophins, as important regulators of neural development, function, and survival, have a therapeutic potential to repair damaged neurons. However, a controlled delivery of therapeutic molecules to injured tissue remains one of the greatest challenges facing the translation of novel drug therapeutics field. This study presents the development of an innovative protein-protein delivery technology of nerve growth factor (NGF) by an electrostatically assembled protein-based (collagen) reservoir system that can be directly injected into the injury site and provide long-term release of the therapeutic. A protein-based biomimetic hollow reservoir system was fabricated using a template method. The capability of neurotrophins to localize in these reservoir systems was confirmed by confocal images of fluorescently labeled collagen and NGF. In addition, high loading efficiency of the reservoir system was proven using ELISA. By comparing release profile from microspheres with varying cross-linking, highly cross-linked collagen spheres were chosen as they have the slowest release rate. Finally, biological activity of released NGF was assessed using rat pheochromocytoma (PC12) cell line and primary rat dorsal root ganglion (DRG) cell bioassay where cell treatment with NGF-loaded reservoirs induced significant neuronal outgrowth, similar to that seen in NGF treated controls. Data presented here highlights the potential of a high capacity reservoir-growth factor technology as a promising therapeutic treatment for neuroregenerative applications and other neurodegenerative diseases.
神经生长因子（NGF）是一种重要的神经营养因子，具有修复受损神经元的治疗潜力。然而，将治疗分子递送到损伤组织仍然是新药物治疗领域转化所面临的最大挑战之一。本研究提出了一种创新的蛋白质-蛋白质递药技术，即通过静电组装的基于蛋白质（胶原）的储库系统来传递神经生长因子（NGF），该系统可直接注射到损伤部位，并提供治疗药物的长期释放。使用模板法制备了基于蛋白质的仿生中空储库系统。通过对荧光标记胶原和 NGF 的共焦图像证实了神经营养因子在这些储库系统中的定位能力。此外，通过 ELISA 证明了储库系统具有较高的载药效率。通过比较具有不同交联度的微球的释放曲线，选择了高度交联的胶原微球，因为它们具有最慢的释放速率。最后，使用大鼠嗜铬细胞瘤（PC12）细胞系和原代大鼠背根神经节（DRG）细胞生物测定评估了释放的 NGF 的生物学活性，结果表明负载 NGF 的储库处理后的细胞诱导了明显的神经元突起生长，类似于用 NGF 处理的对照组。这里呈现的数据突出了高容量储库-生长因子技术作为神经再生应用和其他神经退行性疾病有前途的治疗方法的潜力。
