Faculty of Dentistry, Restorative Dental Sciences, The University of Hong Kong, Hong Kong, China.
Shenzhen Stomatology Hospital (Pingshan), Southern Medical University, Shenzhen, China.
Tissue Eng Part A. 2024 Sep;30(17-18):563-576. doi: 10.1089/ten.TEA.2023.0330. Epub 2024 Jun 10.
Ischemic stroke is a devastating medical condition with poor prognosis due to the lack of effective treatment modalities. Transplantation of human neural stem cells or primary neural cells is a promising treatment approach, but this is hindered by limited suitable cell sources and low expansion capacity. This study aimed (1) use small molecules (SM) to reprogram gingival mesenchymal stem cells (GMSCs) commitment to the neural lineage cells , and (2) use hyaluronic acid (HA) hydrogel scaffolds seeded with GMSCs-derived neural lineage cells to treat ischemic stroke . Neural induction was carried out with a SM cocktail-based one-step culture protocol over a period of 24 h. The induced cells were analyzed for expression of neural markers with immunocytochemistry and quantitative real-time polymerase chain reaction (qRT-PCR). The Sprague-Dawley (SD) rats ( = 100) were subjected to the middle cerebral artery occlusion (MCAO) reperfusion ischemic stroke model. Then, after 8 days post-MCAO, the modeled rats were randomly assigned to six study groups ( = 12 per group): (1) GMSCs, (2) GMSCs-derived neural lineage cells, (3) HA and GMSCs-derived neural lineage cells, (4) HA, (5) PBS, and (6) sham transplantation control, and received their respective transplantation. Evaluation of post-stroke recovery were performed by behavioral tests and histological assessments. The morphologically altered nature of neural lineages has been observed of the GMSCs treated with SMs compared to the untreated controls. As shown by the qRT-PCR and immunocytochemistry, SMs further significantly enhanced the expression level of neural markers of GMSCs as compared with the untreated controls (all < 0.05). Intracerebral injection of self-assembling HA hydrogel carrying GMSCs-derived neural lineage cells promoted the recovery of neural function and reduced ischemic damage in rats with ischemic stroke, as demonstrated by histological examination and behavioral assessments (all < 0.05). In conclusion, the SM cocktail significantly enhanced the differentiation of GMSCs into neural lineage cells. The HA hydrogel was found to facilitate the proliferation and differentiation of GMSCs-derived neural lineage cells. Furthermore, HA hydrogel seeded with GMSCs-derived neural lineage cells could promote tissue repair and functional recovery in rats with ischemic stroke and may be a promising alternative treatment modality for stroke.
缺血性中风是一种预后不良的破坏性医学病症，由于缺乏有效的治疗方法。移植人神经干细胞或原代神经细胞是一种很有前途的治疗方法，但这受到合适细胞来源有限和扩增能力低的限制。本研究旨在：(1) 使用小分子 (SM) 将牙龈间充质干细胞 (GMSC) 重编程为神经谱系细胞，(2) 使用透明质酸 (HA) 水凝胶支架种植 GMSC 衍生的神经谱系细胞治疗缺血性中风。通过基于 SM 鸡尾酒的一步培养方案在 24 小时内进行神经诱导。通过免疫细胞化学和实时定量聚合酶链反应 (qRT-PCR) 分析诱导细胞的神经标志物表达。将 Sprague-Dawley (SD) 大鼠（=100）进行大脑中动脉闭塞 (MCAO) 再灌注缺血性中风模型。然后，在 MCAO 后 8 天，将建模大鼠随机分为六个研究组（每组=12）：(1) GMSC，(2) GMSC 衍生的神经谱系细胞，(3) HA 和 GMSC 衍生的神经谱系细胞，(4) HA，(5) PBS 和 (6) 假手术对照，并接受各自的移植。通过行为测试和组织学评估评估中风后恢复情况。与未处理的对照组相比，用 SM 处理的 GMSC 表现出神经谱系形态改变。如 qRT-PCR 和免疫细胞化学所示，与未处理的对照组相比，SM 进一步显著增强了 GMSC 的神经标志物表达水平（均 <0.05）。载有 GMSC 衍生的神经谱系细胞的自组装 HA 水凝胶的脑内注射促进了缺血性中风大鼠神经功能的恢复和缺血损伤的减少，如组织学检查和行为评估所示（均 <0.05）。总之，SM 鸡尾酒显著增强了 GMSC 向神经谱系细胞的分化。发现 HA 水凝胶有利于 GMSC 衍生的神经谱系细胞的增殖和分化。此外，载有 GMSC 衍生的神经谱系细胞的 HA 水凝胶可促进缺血性中风大鼠的组织修复和功能恢复，可能是中风的一种有前途的替代治疗方法。
