Functionalised microneedles for enhanced neuronal adhesion

摘要

Efficient functional coupling of neuronal cells and electronic sensors could result in hybrid bidirectional communication between neurons and computers (L.J. Breckenridge et al., Advantages of using microfabricated extracellular electrodes for in vitro neuronal recordings, J. Neurosci Res. 42 (1995), pp. 266-276; G. Zeck and P. Fromherz, Noninvasive neuroelectronic interfacing with synaptically connected snail neurons immobilised on a semiconductor chip, PNAS 98 (2001), pp. 10457-10462). Such systems could enable us to gain insight into the mechanisms of neuro-degenerative diseases like Parkinson's and Alzheimer's disease in vitro or could be used to improve the function and efficiency of devices used in vivo, for example in Deep Brain Stimulation devices that are already used in the treatment of Parkinson's disease. One of the major challenges for the development of reliable neuro-electronic systems is to perform extracellular recordings of action potentials with a high signal-to-noise ratio. The poor quality of these recordings is caused by the culture medium, which is present in the cleft between the cell membrane and the sensor surface (P. Fromherz, Neuroelectronic interfacing: semiconductor chips with ion channels, nerve cells and brain, in Nanoelectronics and Information Technology, R. Waser, ed., Wiley-VCH, Berlin, 2003, pp. 781-810; G. Zeck and P. Fromherz, Noninvasive neuroelectronic interfacing with synaptically connected snail neurons immobilised on a semiconductor chip, PNAS 98 (2001), pp. 10457-10462). In this article, we describe a method allowing a reduction of the distance between the membrane and the surface by combining surface chemistry and topography. We have developed a specialised surface chemistry, based on small laminin-derived peptides, which applied onto the topographical structures, triggers their engulfment by the cell membrane in a phagocytosis-like event. In the phagocytotic pit, the distance between the cell membrane and the sensor surface is believed to be minimal. We describe the surface chemistry used for the controlled immobilisation of the small peptides on the surface of the needle-like structures that are manufactured on the surface of electronic devices. PC12 neuroblastoma cells and genetically-modified HeLa cells have been used to investigate the interaction between the cell membrane and the peptide functionalised topographical structures. The membrane-surface interaction was examined by means of electron microscopy and fluorescence microscopy.