Martín Daniel, Bocio-Nuñez J, Scagliusi Santiago F, Pérez Pablo, Huertas Gloria, Yúfera Alberto, Giner Mercè, Daza Paula
Electronics Technology Department, Universidad de Sevilla, Seville, Spain.
Microelectronics Institute of Seville, Universidad de Sevilla, Seville, Spain.
J Biol Eng. 2022 Oct 13;16(1):27. doi: 10.1186/s13036-022-00306-8.
Electrical stimulation is a novel tool to promote the differentiation and proliferation of precursor cells. In this work we have studied the effects of direct current (DC) electrical stimulation on neuroblastoma (N2a) and osteoblast (MC3T3) cell lines as a model for nervous and bone tissue regeneration, respectively. We have developed the electronics and encapsulation of a proposed stimulation system and designed a setup and protocol to stimulate cell cultures.
Cell cultures were subjected to several assays to assess the effects of electrical stimulation on them. N2a cells were analyzed using microscope images and an inmunofluorescence assay, differentiated cells were counted and neurites were measured. MC3T3 cells were subjected to an AlamarBlue assay for viability, ALP activity was measured, and a real time PCR was carried out.
Our results show that electrically stimulated cells had more tendency to differentiate in both cell lines when compared to non-stimulated cultures, paired with a promotion of neurite growth and polarization in N2a cells and an increase in proliferation in MC3T3 cell line.
These results prove the effectiveness of electrical stimulation as a tool for tissue engineering and regenerative medicine, both for neural and bone injuries. Bone progenitor cells submitted to electrical stimulation have a higher tendency to differentiate and proliferate, filling the gaps present in injuries. On the other hand, neuronal progenitor cells differentiate, and their neurites can be polarized to follow the electric field applied.
电刺激是促进前体细胞分化和增殖的一种新型工具。在本研究中,我们分别以神经母细胞瘤(N2a)细胞系和成骨细胞(MC3T3)细胞系作为神经组织和骨组织再生的模型,研究了直流电(DC)刺激对它们的影响。我们开发了一种拟议的刺激系统的电子设备和封装,并设计了一种装置和方案来刺激细胞培养物。
对细胞培养物进行了多项检测,以评估电刺激对它们的影响。使用显微镜图像和免疫荧光检测分析N2a细胞,对分化细胞进行计数并测量神经突。对MC3T3细胞进行AlamarBlue活力检测,测量碱性磷酸酶(ALP)活性,并进行实时聚合酶链反应(PCR)。
我们的结果表明,与未刺激的培养物相比,电刺激的细胞在两种细胞系中都更倾向于分化,同时促进了N2a细胞的神经突生长和极化,并增加了MC3T3细胞系的增殖。
这些结果证明了电刺激作为组织工程和再生医学工具对于神经和骨损伤的有效性。接受电刺激的骨祖细胞具有更高的分化和增殖倾向,可填补损伤中存在的间隙。另一方面,神经祖细胞分化,其神经突可极化以跟随施加的电场。