Akaranuchat Nutthawut, Chruewkamlow Nuttapol, Sathan-Ard Chutipon, Phutthakunphithak Phakawan, Tapechum Sompol, Ruangsetakit Chanean, Sermsathanasawadi Nuttawut
Division of Plastic and Reconstructive Surgery, Department of Surgery, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Siriraj Center for Regenerative Medicine, Research Department, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
PLoS One. 2025 Apr 16;20(4):e0321457. doi: 10.1371/journal.pone.0321457. eCollection 2025.
This study aimed to assess the efficacy of Quality and Quantity media-cultured mononuclear cells (QQ-MNCs) for promoting nerve regeneration in a mouse sciatic nerve transection model. Human peripheral blood mononuclear cells (PB-MNCs) and QQ-MNCs derived from healthy volunteers were used/compared. The left sciatic nerve was surgically transected in 27 mice. After complete nerve transection was confirmed, end-to-end direct epineurial nerve repair was performed using 9-0 nylon. Fibrin glue was applied to the tissue around the injury site to limit diffusion of the study treatment followed by application of 0.5 ml phosphate buffered saline (PBS) or PB-MNCs (2x106 cells) or QQ-MNCs (2x106 cells) to the injury site. The skin was then closed using 6-0 nylon. Histomorphology, immunohistochemistry, electrophysiologic examination, and functional assessment were evaluated at 12-weeks followed by euthanasia and subsequent harvesting of the left sciatic nerves and the left and right gastrocnemius muscles for examination. QQ-MNCs mice exhibited significant improvement in all histomorphologic parameters (axon fiber diameter, myelin thickness, percentage of nerve density) and immunohistochemistry assays (S100, SOX10, GFAP, neurofilament, IL-1β, VEGF, anti-HNA, TNF-α, vWF) compared to PBS mice (all p < 0.05). QQ-MNCs mice also had a significantly higher Basso Mouse Scale score compared to PBS mice (p = 0.018). The percentage of nerve density adjacent to the injury site was significantly higher in QQ-MNCs mice than in PB-MNCs mice (p = 0.049). IL-1β expression was significantly lower in QQ-MNCs mice than in PB-MNCs mice (p = 0.01). QQ-MNCs mice demonstrated significantly better functional and histomorphologic outcomes of nerve regeneration compared to PB-MNCs mice and PBS mice.
本研究旨在评估质量和数量培养基培养的单核细胞（QQ-MNCs）在小鼠坐骨神经横断模型中促进神经再生的疗效。使用/比较了源自健康志愿者的人外周血单核细胞（PB-MNCs）和QQ-MNCs。对27只小鼠的左侧坐骨神经进行手术横断。在确认神经完全横断后，使用9-0尼龙线进行端到端直接神经外膜修复。将纤维蛋白胶应用于损伤部位周围的组织，以限制研究治疗的扩散，随后将0.5毫升磷酸盐缓冲盐水（PBS）或PB-MNCs（2×10⁶个细胞）或QQ-MNCs（2×10⁶个细胞）应用于损伤部位。然后用6-0尼龙线缝合皮肤。在12周时评估组织形态学、免疫组织化学、电生理检查和功能评估，随后实施安乐死，并随后采集左侧坐骨神经以及左、右腓肠肌进行检查。与PBS小鼠相比，QQ-MNCs小鼠在所有组织形态学参数（轴突纤维直径、髓鞘厚度、神经密度百分比）和免疫组织化学检测（S100、SOX10、GFAP、神经丝、IL-1β、VEGF、抗HNA、TNF-α、vWF）方面均表现出显著改善（所有p<0.05）。与PBS小鼠相比，QQ-MNCs小鼠的巴索小鼠量表评分也显著更高（p=0.018）。QQ-MNCs小鼠损伤部位相邻的神经密度百分比显著高于PB-MNCs小鼠（p=0.049）。QQ-MNCs小鼠的IL-1β表达显著低于PB-MNCs小鼠（p=0.01）。与PB-MNCs小鼠和PBS小鼠相比，QQ-MNCs小鼠在神经再生的功能和组织形态学结果方面表现明显更好。
