de Castro Mateus Vidigal, da Silva Moníze Valéria Ramos, Chiarotto Gabriela Bortolança, Santana Maria Helena Andrade, Luzo Ângela Cristina Malheiros, Kyrylenko Sergiy, de Oliveira Alexandre Leite Rodrigues
Department of Structural and Functional Biology, Institute of Biology, University of Campinas, Campinas, São Paulo, Brazil.
University Center of Herminio Ometto Foundation, Araras, São Paulo, Brazil.
Stem Cells Int. 2020 Oct 29;2020:8834360. doi: 10.1155/2020/8834360. eCollection 2020.
Dorsal root rhizotomy (DRZ) is currently considered an untreatable injury, resulting in the loss of sensitive function and usually leading to neuropathic pain. In this context, we recently proposed a new surgical approach to treat DRZ that uses platelet-rich plasma (PRP) gel to restore the spinal reflex. Success was correlated with the reentry of primary afferents into the spinal cord. Here, aiming to enhance previous results, cell therapy with bioengineered human embryonic stem cells (hESCs) to overexpress fibroblast growth factor 2 (FGF2) was combined with PRP. For these experiments, adult female rats were submitted to a unilateral rhizotomy of the lumbar spinal dorsal roots, which was followed by root repair with PRP gel with or without bioengineered hESCs. One week after DRZ, the spinal cords were processed to evaluate changes in the glial response (GFAP and Iba-1) and excitatory synaptic circuits (VGLUT1) by immunofluorescence. Eight weeks postsurgery, the lumbar intumescences were processed for analysis of the repaired microenvironment by transmission electron microscopy. Spinal reflex recovery was evaluated by the electronic Von Frey method for eight weeks. The transcript levels for human FGF2 were over 37-fold higher in the induced hESCs than in the noninduced and the wildtype counterparts. Altogether, the results indicate that the combination of hESCs with PRP gel promoted substantial and prominent axonal regeneration processes after DRZ. Thus, the repair of dorsal roots, if done appropriately, may be considered an approach to regain sensory-motor function after dorsal root axotomy.
背根切断术(DRZ)目前被认为是一种无法治疗的损伤,会导致感觉功能丧失,通常还会引发神经性疼痛。在此背景下,我们最近提出了一种治疗DRZ的新手术方法,即使用富含血小板血浆(PRP)凝胶来恢复脊髓反射。成功与初级传入神经重新进入脊髓有关。在此,为了强化之前的结果,将生物工程化人胚胎干细胞(hESCs)过表达成纤维细胞生长因子2(FGF2)的细胞疗法与PRP相结合。对于这些实验,成年雌性大鼠接受了腰脊髓背根的单侧切断术,随后用含或不含生物工程化hESCs的PRP凝胶进行神经根修复。DRZ后一周,通过免疫荧光处理脊髓,以评估胶质细胞反应(GFAP和Iba - 1)和兴奋性突触回路(VGLUT1)的变化。术后八周,通过透射电子显微镜处理腰膨大,以分析修复后的微环境。通过电子冯弗里方法评估脊髓反射恢复情况,为期八周。诱导后的hESCs中人类FGF2的转录水平比未诱导的和野生型对应物高出37倍以上。总体而言,结果表明hESCs与PRP凝胶的组合在DRZ后促进了大量且显著的轴突再生过程。因此,如果操作得当,背根修复可被视为背根轴突切断术后恢复感觉运动功能的一种方法。