Genchi Giada Graziana, Ciofani Gianni, Polini Alessandro, Liakos Ioannis, Iandolo Donata, Athanassiou Athanassia, Pisignano Dario, Mattoli Virgilio, Menciassi Arianna
Scuola Superiore Sant'Anna, The BioRobotics Institute, Viale Rinaldo Piaggio 34, 56025, Pontedera, (Pisa), Italy; Istituto Italiano di Tecnologia, Center for MicroBioRobotics @SSSA, Viale Rinaldo Piaggio 34, 56025, Pontedera, (Pisa), Italy.
J Tissue Eng Regen Med. 2015 Feb;9(2):151-61. doi: 10.1002/term.1623. Epub 2012 Oct 22.
In the last decade, the importance of topographic properties of extracellular environments has been shown to be essential to addressing cell response, especially when replacing damaged tissues with functional constructs obtained in vitro. In the current study, densely packed sub-micron poly(3-hydroxybutyrate) (PHB) fibres were electrospun with random and parallel orientations. PC12 pheochromocytoma cells that mimic central dopaminergic neurons and represent a model for neuronal differentiation were cultured on collagen-coated fibres to evaluate cell response dependence on substrate topography. Cell adhesion, viability and proliferation, as well as dopamine production were evaluated after three days since seeding. Cell differentiation was examined in terms of neurite number, orientation and length 6 days after administration of nerve growth factor (NGF). Results showed that proliferating PC12 cells secreted a higher quantity of dopamine on fibres with respect to control cultures and as a result, a possible use of PHB fibres was considered for cell transplantation in the central nervous system when local production of dopamine is impaired. Differentiated PC12 cells were characterized by highly aligned and longer neurites on parallel PHB fibres with respect to random fibres, thereby demonstrating the suitability of parallel PHB fibres for further studies in peripheral nervous system regeneration.
在过去十年中,细胞外环境的拓扑特性对于解决细胞反应的重要性已被证明至关重要,特别是在用体外获得的功能性构建体替代受损组织时。在当前研究中,将紧密堆积的亚微米聚(3-羟基丁酸酯)(PHB)纤维以随机和平行取向进行电纺丝。在胶原包被的纤维上培养模拟中枢多巴胺能神经元并代表神经元分化模型的PC12嗜铬细胞瘤细胞,以评估细胞反应对底物拓扑结构的依赖性。接种三天后评估细胞粘附、活力和增殖以及多巴胺产生。在给予神经生长因子(NGF)6天后,从神经突数量、取向和长度方面检查细胞分化。结果表明,与对照培养物相比,增殖的PC12细胞在纤维上分泌更高量的多巴胺,因此,当多巴胺的局部产生受损时,考虑将PHB纤维用于中枢神经系统的细胞移植。与随机纤维相比,分化的PC12细胞在平行PHB纤维上具有高度排列且更长的神经突,从而证明平行PHB纤维适用于外周神经系统再生方面的进一步研究。