Jung Seunghyun, Harris Nathaniel, Niyonshuti Isabelle I, Jenkins Samir V, Hayar Abdallah M, Watanabe Fumiya, Jamshidi-Parsian Azemat, Chen Jingyi, Borrelli Michael J, Griffin Robert J
Department of Physiology and Cell Biology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA.
Department of Radiation Oncology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA.
Nanomaterials (Basel). 2021 May 4;11(5):1216. doi: 10.3390/nano11051216.
Strategies to increase the proportion of neural stem cells that differentiate into neurons are vital for therapy of neurodegenerative disorders. In vitro, the extracellular matrix composition and topography have been found to be important factors in stem cell differentiation. We have developed a novel artificial extracellular matrix (aECM) formed by attaching gold nanocages (AuNCs) to glass coverslips. After culturing rat neural stem cells (rNSCs) on these gold nanocage-coated surfaces (AuNC-aECMs), we observed that 44.6% of rNSCs differentiated into neurons compared to only 27.9% for cells grown on laminin-coated glass coverslips. We applied laser irradiation to the AuNC-aECMs to introduce precise amounts of photothermally induced heat shock in cells. Our results showed that laser-induced thermal stimulation of AuNC-aECMs further enhanced neuronal differentiation (56%) depending on the laser intensity used. Response to these photothermal effects increased the expression of heat shock protein 27, 70, and 90α in rNSCs. Analysis of dendritic complexity showed that this thermal stimulation promoted neuronal maturation by increasing dendrite length as thermal dose was increased. In addition, we found that cells growing on AuNC-aECMs post laser irradiation exhibited action potentials and increased the expression of voltage-gated Na+ channels compared to laminin-coated glass coverslips. These results indicate that the photothermal response induced in cells growing on AuNC-aECMs can be used to produce large quantities of functional neurons, with improved electrochemical properties, that can potentially be transplanted into a damaged central nervous system to provide replacement neurons and restore lost function.
提高分化为神经元的神经干细胞比例的策略对于神经退行性疾病的治疗至关重要。在体外,已发现细胞外基质组成和拓扑结构是干细胞分化的重要因素。我们开发了一种通过将金纳米笼(AuNCs)附着在玻璃盖玻片上形成的新型人工细胞外基质(aECM)。在这些金纳米笼包被的表面(AuNC-aECMs)上培养大鼠神经干细胞(rNSCs)后,我们观察到44.6%的rNSCs分化为神经元,而在层粘连蛋白包被的玻璃盖玻片上生长的细胞只有27.9%分化为神经元。我们对AuNC-aECMs施加激光照射,以在细胞中引入精确量的光热诱导热休克。我们的结果表明,根据所使用的激光强度,激光诱导的AuNC-aECMs热刺激进一步增强了神经元分化(56%)。对这些光热效应的反应增加了rNSCs中热休克蛋白27、70和90α的表达。树突复杂性分析表明,随着热剂量增加,这种热刺激通过增加树突长度促进了神经元成熟。此外,我们发现,与层粘连蛋白包被的玻璃盖玻片相比,激光照射后在AuNC-aECMs上生长的细胞表现出动作电位,并增加了电压门控Na+通道的表达。这些结果表明,在AuNC-aECMs上生长的细胞中诱导的光热反应可用于产生大量具有改善的电化学特性的功能性神经元,这些神经元可能被移植到受损的中枢神经系统中以提供替代神经元并恢复丧失的功能。