Institute of New Frontier Research, Hallym University College of Medicine, Chuncheon, Korea.
Department of Neurosurgery, Yonsei University Wonju College of Medicine, Wonju, Korea.
Acta Biomater. 2023 Sep 1;167:335-347. doi: 10.1016/j.actbio.2023.06.027. Epub 2023 Jun 23.
There are no effective clinically applicable treatments for neuronal dysfunction after mild traumatic brain injury (TBI). Here, we evaluated the therapeutic effect of a new delivery method of mouse neural stem cell (mNSC) spheroids using a hydrogel, in terms of improvement in damaged cortical lesions and cognitive impairment after mild TBI.
mNSCs were isolated from the subventricular zone and subgranular zone by a hydrogel-based culture system. GFP-transduced mNSCs were generated into spheroids and wrapped into a sheet for transplantation. Male C57BL/6J mice were randomly divided into four groups: sham operation, TBI, TBI with mNSC spheroids, and TBI with mNSC spheroid sheet transplantation covering the damaged cortex. Histopathological and immunohistochemical features and cognitive function were evaluated 7, 14, and 28 days after transplantation following TBI.
Hydrogel-based culture systems and mNSC isolation were successfully established from the adult mice. Essential transcription factors for NSCs, such as SOX2, PAX6, Olig2, nestin, and doublecortin (DCX), were highly expressed in the mNSCs. A transplanted hydrogel-based mNSC spheroid sheet showed good engraftment and survival ability, differentiated into TUJ1-positive neurons, promoted angiogenesis, and reduced neuronal degeneration. Also, TBI mice treated with mNSC spheroid sheet transplantation exhibited a significantly increased preference for a new object, suggesting improved cognitive function compared to the mNSC spheroids or no treatment groups.
Transplantation with a hydrogel-based mNSC spheroid sheet showed engraftment, migration, and stability of delivered cells in a hostile microenvironment after TBI, resulting in improved cognitive function via reconstruction of the damaged cortex.
This study presents the therapeutic effect of a new delivery method of mouse neural stem cells spheroids using a hydrogel, in terms of improvement in damaged cortical lesions and cognitive impairment after traumatic brain injury. Collagen/fibrin hydrogel allowed long-term survival and migratory ability of NSCs spheroids. Furthermore, transplanted hydrogel-based mNSCs spheroids sheet showed good engraftment, migration, and stability of delivered cells in a hostile microenvironment, resulting in reconstruction of the damaged cortex and improved cognitive function after TBI. Therefore, we suggest that a hydrogel-based mNSCs spheroids sheet could help to improve cognitive impairment after TBI.
目前对于轻度创伤性脑损伤（TBI）后神经元功能障碍尚无有效的临床应用治疗方法。在这里，我们评估了使用水凝胶的新型小鼠神经干细胞（mNSC）球体传递方法的治疗效果，以改善轻度 TBI 后受损皮质病变和认知障碍。
通过基于水凝胶的培养系统从侧脑室下区和颗粒下区分离 mNSC。将 GFP 转导的 mNSC 生成球体并包裹成薄片用于移植。雄性 C57BL/6J 小鼠随机分为四组：假手术、TBI、TBI 加 mNSC 球体和 TBI 加覆盖受损皮质的 mNSC 球体薄片移植。TBI 后 7、14 和 28 天，评估移植后组织病理学和免疫组织化学特征以及认知功能。
成功地从成年小鼠中建立了基于水凝胶的培养系统和 mNSC 分离。NSC 的必需转录因子，如 SOX2、PAX6、Olig2、巢蛋白和双皮质素（DCX），在 mNSC 中高度表达。移植的基于水凝胶的 mNSC 球体薄片显示出良好的植入和存活能力，分化为 TUJ1 阳性神经元，促进血管生成，并减少神经元变性。此外，与 mNSC 球体或未治疗组相比，接受 mNSC 球体薄片移植的 TBI 小鼠对新物体的偏好明显增加，表明认知功能得到改善。
在 TBI 后，基于水凝胶的 mNSC 球体薄片的移植显示出植入、迁移和递送细胞在恶劣微环境中的稳定性，通过受损皮质的重建改善认知功能。
本研究提出了一种新的利用水凝胶传递小鼠神经干细胞球体的方法的治疗效果，可改善创伤性脑损伤后受损皮质病变和认知障碍。胶原/纤维蛋白水凝胶允许 NSCs 球体的长期存活和迁移能力。此外，移植的基于水凝胶的 mNSC 球体薄片在恶劣的微环境中显示出良好的植入、迁移和递送细胞的稳定性，导致 TBI 后受损皮质的重建和认知功能的改善。因此，我们认为基于水凝胶的 mNSC 球体薄片有助于改善 TBI 后的认知障碍。
