Department of Regenerative Medicine, Maria Sklodowska-Curie National Research Institute of Oncology, W.K. Roentgena 5, 02-781 Warsaw, Poland.
Department of Regenerative Medicine, Maria Sklodowska-Curie National Research Institute of Oncology, W.K. Roentgena 5, 02-781 Warsaw, Poland.
Tissue Cell. 2022 Oct;78:101899. doi: 10.1016/j.tice.2022.101899. Epub 2022 Aug 20.
The development of tissue-engineered scaffolds with electrical properties is the primary motivation of novel regenerative medicine. Electroconductive scaffolds are designed to mimic the injured tissue environment's electrical properties and regulate cellular behavior - growth, proliferation, and differentiation - that could stimulate the injured nerve's regeneration.
We fabricated dedicated electroconductive scaffolds and customized an appropriate device with an external current supply to expose cells on the scaffold to electrical stimulation (ES). Next, we isolated rat adipose-derived stem cells (ASCs) and performed in vitro experiments that combine cells, an electroconductive scaffold, NGF (nerve growth factor), and ES (90 mV/mm, constant, for four days). Finally, we checked cellular activity as proliferation, viability, morphology, the neurogenic differentiation potential of ASCs, cell alignment, and karyotype.
We observed that the electrical stimulation did not change the viability and chromosome stability of rat ASCs, but altered slightly proliferation compared to non-stimulated cells. The combined effect of a scaffold, NGF, and ES caused morphology changes and enhancement of ASCs neuronal differentiation as indicated in βIII-tubulin expression, actin organization, and upregulation of neurogenic gene expression.
We developed an electroconductive scaffold and customized device for in vitro study with many experimental variants. Based on our results, we presumed that the established study scheme - including an electroconductive scaffold, NGF and ES - is biocompatible and could guide ASCs to differentiate in neurogenic lineage, thus may be potentially applied in nerve injury regeneration.
具有电性能的组织工程支架的开发是新型再生医学的主要动力。导电支架旨在模拟受伤组织环境的电性能,并调节细胞行为——生长、增殖和分化——这可以刺激受伤神经的再生。
我们制造了专用的导电支架,并定制了一个带有外部电流供应的合适设备,以使支架上的细胞暴露于电刺激(ES)下。接下来,我们分离大鼠脂肪来源干细胞(ASCs),并进行体外实验,将细胞、导电支架、NGF(神经生长因子)和 ES(90 mV/mm,恒定,持续四天)结合起来。最后,我们检查了细胞活性,如增殖、活力、形态、ASCs 的神经分化潜能、细胞排列和核型。
我们观察到电刺激没有改变大鼠 ASCs 的活力和染色体稳定性,但与未受刺激的细胞相比,增殖略有改变。支架、NGF 和 ES 的联合作用导致细胞形态发生变化,并增强了 ASCs 的神经元分化,表现为βIII-微管蛋白表达、肌动蛋白组织和神经基因表达的上调。
我们开发了一种导电支架和定制的设备,用于具有多种实验变体的体外研究。基于我们的结果,我们推测所建立的研究方案——包括导电支架、NGF 和 ES——是生物相容的,可以指导 ASCs 向神经谱系分化,因此可能潜在地应用于神经损伤再生。