Paes Sabrina Moreira, Castro Mateus Vidigal de, Barbosa Rafael Maza, Politti Cartarozzi Luciana, Coser Lilian de Oliveira, Kempe Paula Regina Gelinski, Decarli Monize Caiado, Moraes Ângela Maria, Barraviera Benedito, Ferreira Júnior Rui Seabra, Oliveira Alexandre Leite Rodrigues de
Laboratory of Nerve Regeneration, Institute of Biology (IB), State University of Campinas (UNICAMP), Campinas, SP, Brazil.
Laboratory of Nerve Regeneration, Institute of Biology (IB), State University of Campinas (UNICAMP), Campinas, SP, Brazil; School of Chemical Engineering, University of Campinas, UNICAMP, Av. Albert Einstein, 500, Brazil.
Brain Res. 2023 Mar 1;1802:148229. doi: 10.1016/j.brainres.2022.148229. Epub 2022 Dec 30.
Spinal cord injuries result in severe neurological deficits and neuronal loss, with poor functional recovery. Mesenchymal stem cells have shown promising results; therefore the present objective of this work was to compare motor recovery after treatment with human dental pulp stem cells (hDPSC) cultivated in monolayer (2D) or as spheroids (3D), following avulsion and reimplantation of spinal motor roots in adult rats. Thus, 72 adult female Lewis rats were divided into 4 groups: avulsion (AV); avulsion followed by reimplantation (AR); avulsion associated with reimplant and 2D cell therapy (AR + 2D), and avulsion associated with reimplant and 3D cell therapy (AR + 3D). The application of the cells in 2D and 3D was performed by microsurgery, with subsequent functional assessment using a walking track test (Catwalk system), immunohistochemistry, neuronal survival, and qRT-PCR in 1-, 4-, and 12-weeks post-injury. The animals in the AR + 2D and AR + 3D groups showed the highest neuronal survival rates, and immunofluorescence revealed downregulation of GFAP, and Iba-1, with preservation of synaptophysin, indicating a reduction in glial reactivity, combined with the maintenance of pre-synaptic inputs. There was an increase in anti-inflammatory (IL-4, TGFβ) and a reduction of pro-inflammatory factors (IL-6, TNFα) in animals treated with reimplantation and hDPSC. As for the functional recovery, in all analyzed parameters, the AR + 2D group performed better and was superior to the avulsion alone. Overall, our results indicate that the 2D and 3D cell therapy approaches provide successful immunomodulation and motor recovery, consistent with advanced therapies after spinal cord injury.
脊髓损伤会导致严重的神经功能缺损和神经元丢失，功能恢复较差。间充质干细胞已显示出有前景的结果；因此，本研究的目的是比较在成年大鼠脊髓运动神经根撕脱并重新植入后，用单层培养（2D）或球体培养（3D）的人牙髓干细胞（hDPSC）治疗后的运动恢复情况。因此，将72只成年雌性Lewis大鼠分为4组：撕脱组（AV）；撕脱后重新植入组（AR）；撕脱并重新植入联合2D细胞治疗组（AR + 2D），以及撕脱并重新植入联合3D细胞治疗组（AR + 3D）。通过显微手术将细胞以2D和3D方式应用，随后在损伤后1周、4周和12周使用步行轨迹测试（Catwalk系统）、免疫组织化学、神经元存活情况和qRT-PCR进行功能评估。AR + 2D组和AR + 3D组的动物显示出最高的神经元存活率，免疫荧光显示胶质纤维酸性蛋白（GFAP）和离子钙结合衔接分子1（Iba-1）下调，同时突触素得以保留，表明胶质细胞反应性降低，同时突触前输入得以维持。在接受重新植入和hDPSC治疗的动物中，抗炎因子（白细胞介素-4、转化生长因子β）增加，促炎因子（白细胞介素-6、肿瘤坏死因子α）减少。至于功能恢复，在所有分析参数中，AR + 2D组表现更好，优于单纯撕脱组。总体而言，我们的结果表明，2D和3D细胞治疗方法能成功实现免疫调节和运动恢复，与脊髓损伤后的先进治疗方法一致。
