Bogomoletz Institute of Physiology NAS of Ukraine, Kyiv 01601, Ukraine; State Institution National Scientific Center the M.D. Strazhesko Institute of Cardiology, Clinical and Regenerative Medicine, NAMS of Ukraine, Kyiv 03680, Ukraine.
Bogomoletz Institute of Physiology NAS of Ukraine, Kyiv 01601, Ukraine.
Exp Neurol. 2023 Oct;368:114497. doi: 10.1016/j.expneurol.2023.114497. Epub 2023 Jul 29.
Currently, several therapeutic methods of treating the effects of spinal cord injury (SCI) are being considered. On the one hand, transplantation of stem cells (SCs), in particular, neural stem/progenitor cells (NSPCs), is promising, as these cells have the potential to differentiate into nervous tissue cells, able to enhance endogenous regeneration and prevent the development of inflammatory processes. On the other hand, it is quite promising to replace the damaged nervous tissue with synthetic matrices, in particular hydrogels, which can create artificial conditions for the regenerative growth of injured nerve fibers through the spinal cord injury area, i.e. stimulate and support axonal regeneration and myelination. In this work, we combined both of these novel approaches by populating (injecting or rehydrating) a heteroporous pHPMA hydrogel (NeuroGel) with murine hippocampal NSPCs. Being inside the hydrogel (10 days of cultivation), NSPCs were more differentiated into neurons: 19.48% ± 1.71% (the NSPCs injection into the hydrogel) and 36.49% ± 4.20% (the hydrogel rehydration in the NSPCs suspension); in control cultures, the level of differentiation in neurons was only 2.40% ± 0.31%. Differentiation of NSPCs into glial cells, in particular into oligodendrocyte progenitor cells, was also observed - 8.89% ± 2.15% and 6.21% ± 0.80% for injection and rehydration variants, respectively; in control - 28.75% ± 2.08%. In the control NSPCs culture, there was a small number of astrocytes - 2.11% ± 0.43%. Inside the hydrogel, NSPCs differentiation in astrocytes was not observed. In vitro data showed that the hydrogel promotes the differentiation of NSPCs into neurons, and inhibits the differentiation into glial cells. And in vivo showed post-traumatic recovery of rat spinal cord tissue after injury followed by implantation of the hydrogel+NSPCs complex (approximately 7 months after SCI). The implant area was closely connected with the recipient tissue, and the recipient cells freely grew into the implant itself. Inside the implant, a formed dense neuronal network was visible. In summary, the results are primarily an experimental ground for further studies of implants based on pHPMA hydrogel with populated different origin SCs, and the data also indicate the feasibility and efficiency of using an integrated approach to reduce possible negative side effects and facilitate the rehabilitation process after a SCI.
目前，人们正在考虑几种治疗脊髓损伤 (SCI) 影响的治疗方法。一方面，干细胞（SCs），特别是神经干细胞/祖细胞（NSPCs）的移植具有很大的前景，因为这些细胞具有分化为神经组织细胞的潜力，能够增强内源性再生并防止炎症过程的发展。另一方面，用合成基质，特别是水凝胶替代受损的神经组织具有很大的前景，因为水凝胶可以通过脊髓损伤区域为受伤神经纤维的再生生长创造人工条件，即刺激和支持轴突再生和髓鞘形成。在这项工作中，我们通过将鼠海马 NSPC 接种（注射或再水合）到杂孔 pHPMA 水凝胶（NeuroGel）中来结合这两种新方法。在水凝胶内（培养 10 天），NSPC 更多地分化为神经元：19.48%±1.71%（将 NSPC 注射到水凝胶中）和 36.49%±4.20%（将水凝胶再水合到 NSPC 悬浮液中）；在对照培养物中，神经元分化水平仅为 2.40%±0.31%。还观察到 NSPC 分化为神经胶质细胞，特别是少突胶质前体细胞，注射和再水化变体分别为 8.89%±2.15%和 6.21%±0.80%；在对照中为 28.75%±2.08%。在对照 NSPC 培养物中，只有少量的星形胶质细胞-2.11%±0.43%。在水凝胶内，未观察到 NSPC 分化为星形胶质细胞。体外数据表明，水凝胶促进 NSPC 向神经元分化，并抑制向神经胶质细胞分化。体内研究表明，在大鼠脊髓损伤后植入水凝胶+NSPC 复合物后（SCI 后约 7 个月），脊髓组织有创伤后恢复。植入物区域与受者组织紧密相连，受者细胞自由生长到植入物本身。在植入物内，可以看到形成的密集神经元网络。总之，这些结果主要是进一步研究基于带有接种不同来源干细胞的 pHPMA 水凝胶的植入物的实验基础，并且数据还表明使用综合方法的可行性和效率，以减少可能的负面副作用并促进 SCI 后的康复过程。
