Aix Marseille Univ, CNRS, INSERM, CIML, Turing Centre for Living Systems, Marseille, France.
Aix Marseille Univ, CNRS, CINaM, Marseille, France.
Sci Rep. 2020 Feb 27;10(1):3581. doi: 10.1038/s41598-020-60333-7.
Understanding how animals respond to injury and how wounds heal remains a challenge. These questions can be addressed using genetically tractable animals, including the nematode Caenorhabditis elegans. Given its small size, the current methods for inflicting wounds in a controlled manner are demanding. To facilitate and accelerate the procedure, we fabricated regular arrays of pyramidal features ("pins") sharp enough to pierce the tough nematode cuticle. The pyramids were made from monocrystalline silicon wafers that were micro-structured using optical lithography and alkaline wet etching. The fabrication protocol and the geometry of the pins, determined by electron microscopy, are described in detail. We also used electron microscopy to characterize the different types of injury caused by these pins. Upon wounding, C. elegans expresses genes encoding antimicrobial peptides. A comparison of the induction of antimicrobial peptide gene expression using traditional needles and the pin arrays demonstrates the utility of this new method.
了解动物如何应对损伤以及伤口如何愈合仍然是一个挑战。这些问题可以使用遗传上可操作的动物来解决,包括秀丽隐杆线虫。由于其体型小,目前以受控方式造成伤口的方法要求很高。为了促进和加速该过程,我们制造了规则的金字塔形特征(“针”)阵列,这些针足够锋利,可以刺穿坚韧的线虫表皮。金字塔由单晶硅片制成,使用光学光刻和碱性湿法刻蚀进行微结构处理。详细描述了制造协议和通过电子显微镜确定的针的几何形状。我们还使用电子显微镜来描述这些针造成的不同类型的损伤。受伤后,秀丽隐杆线虫表达编码抗菌肽的基因。使用传统的针和针阵列比较抗菌肽基因表达的诱导,证明了这种新方法的实用性。