臺灣博碩士論文加值系統

神經網路電生理特性描述對於了解功能性神經網路行為扮演其重要角色。以微電極陣列為主的電阻抗量測與膜外電生理紀錄平台，藉由穩定神經細胞培養技術，可提供動態與長期觀測神經網路行為觀測。此研究目的為利用微電極陣列平台來比較體外培養與活體三維神經網路之電生理特性，並利用此平台之高度整合性，於二維體外培養神經網路下建立體外培養缺血缺氧中風與阿茲海默症等神經疾病模型應用。
為架構出三維體外神經網路，首先將大腦皮質神經細胞培養於Matrigel介質中，生長成具有不同深度之三維神經突觸分佈，且具有與二維體外細胞培養相近細胞存活率。於電阻抗量測結果中，神經網路成長與其電阻性成分具備漸增相關性。於膜外神經電生理紀錄結果中，可藉由神經活化模式與神經活化分布分析，提供三維神經網路之功能性證明。為進一步於複雜腦部組織中進行體內植入電生理訊號量測，此研究利用SU-8設計並製作出體內植入彈性探針，於體外與體內神經電生理紀錄中顯示出比傳統硬式神經植入探針更好的組織適應性。且藉由一連串機械與生物毒性測試: 體外機械性質、體外生物毒性測試、活體電生理訊號紀錄與活體免疫螢光染色結果可得到，SU-8彈性神經植入探針具有足夠的機械強度來進行手術植入，且由於其彈性機械性質可達到減少植入免疫反應，進一步得到高於七以上之高品質訊雜比神經訊號。
於神經疾病探討研究中，我們建立可以用來探討神經病理機制之體外神經疾病模型，如體外缺血性中風與近似阿茲海默症之Tau蛋白質異常沉積疾病模型。於體外缺血性中風模型建立探討中，首先藉由微循環環境產生缺血缺氧性中風，利用電阻抗量測於正常與受到生長素介質保護下之大腦皮質細胞進行電阻抗量測。於生物性測試證據中，經由50與100 ng/mL生長素介質保護之大腦皮質細胞顯示出較高的細胞存活率，且於粒線體膜電位表現中無過極化現象之產生。於電阻抗量測結果中，大腦皮質神經細胞經過缺血缺氧後，電阻抗值降低直接反應出細胞貼附性減少。但於50 and 100 ng/mL生長素介質保護之大腦皮質細胞電阻抗量測結果可發現，經過缺血缺氧實驗後，其阻抗值呈現緩慢減少到進行缺血缺氧實驗前的50%，此結果明顯表示，50 and 100 ng/mL生長素介質保護大腦皮質細胞具備較佳細胞貼附。此多時間點連續電阻抗量測結果證明，利用整合微循環系統微電極陣列平台，可於缺血缺氧中風模型下進行即時觀測，且可觀察大腦皮質細胞由生長素介質保護下之效果。
為建立體外阿茲海默症模型，此研究設計以神經突觸連結之健康與疾病大腦皮質神經元族群，應用於了解疾病傳遞之結構與功能差異。結構性資料可藉由螢光影像取得過磷酸化蛋白質tau與神經骨架變化，功能性資料則由膜外電紀錄計算其神經自發性動作電位數量比值與神經自發性大量活動數值比。於結構性結果中觀察得知，於疾病神經元族群中經由okadaic-acid誘發之過磷酸化蛋白質tau透過神經突觸連結區域，進一步對於健康神經元族群產生影響。此延遲性之神經分支退化現象可由免疫螢光影像證明其產生疾病傳遞現象。於功能性結果中觀察得知，其產生出先於結構性改變(神經分支退化)之直接反應電生理形態改變 (神經自發性動作電位數量比值降低)。利用此體外模擬疾病傳遞模型，可進一步提供挑選合適的疾病治療方法或標靶機制選擇的驗證系統。
整體來說。此研究提出之微電極陣列平台可提供活體/體外培養之三維神經網路電生理特性描述，且利用高度整合性之病態生理模型建立特殊神經疾病模擬應用。於此研究成果中得知，藉由普通生理或病理環境下之神經元培養模型可提供一個探討三維神經網路之特性描述，且呈現出其應用於神經疾病治療機制探討的重要性。

Electrophysiological characterization of neuronal networks plays important roles on understanding functional network behaviors. Microelectrode arrays (MEAs)-based platform with impedimetric sensing and extracellular recording could not only monitoring the onset dynamic neuronal network behavior but also with highly feasibility working as long-term observation tool thorough stable cell culture environment. The aims of this study are utilizing MEAs-based platform for comparing fundamental electrophysiological properties of in vitro/ in vivo 3D neuronal network. It also proven with the capacities works as in vitro 2D neurological disease models for ischemic stroke and Alzheimer’s disease (AD) model for pathological mimicking condition.
For constructing 3D neuronal network, cortical neuronal cells were cultured on Matrigel which could grow in a 3D distribution with neurite extension in different depths and exhibit comparable cell survival rate to 2D neuronal culture. The impedimetric measurement was applied to monitor the neuronal growth which was correlated to gradual increase of resistive component of impedance. The neuronal firing pattern exhibited typical spike waveform. The spike raster plot also proved the functional activity of 3D neuronal network. For In-vivo recording, a flexible neuroprobe using SU-8 was designed and fabricated as implant device for complex brain tissue. In vitro and in vivo electrical sensing showed that the improved tissue compatibility compared to that of the traditional rigid neuroprobe. The validation of neuroprobe was achieved by in vitro mechanical and cytotoxicity tests as well as in vivo neural recording and immunohistological staining. Our studies showed that SU-8 neuroprobe possessed enough stress for pentrated into brain tissue and remained flexibility to comply micro-movement of soft tissue with minor immune responses. The proposed SU-8 neuroprobe can achieve in vivo electrophysiological recordings at a signal-noise-ratio of greater than 7.
The in vitro neurological disease models provide an opportunity for getting insight into the pathological mechanism of diseased neuronal networks including ischemic stroke model and AD-like tauopathy model in this study. For the in vitro ischemic stroke model, we utilized impedimetric sensing technique for monitoring the time-course impedimetric changes in normal and insulin-like growth factor 1 (IGF-1)-protected cortical neurons under the ischemic insult of oxygen glucose deprivation (OGD) created in a micro-perfusion environment. From biological evidences, the cortical neurons treated with 50 and 100 ng/mL IGF-1 showed higher survival rates and no occurrence of the hyperpolarization of MMP during the re-oxygenation period. The impedimetric sensing results demonstrated that the measured impedance of cortical neurons decreased due to cell detachment under the insult of OGD. The measured impedance of IGF-1-protected cortical neurons slowly decreased to about 50% of the original value compared to saline control which indicates improved cell adhesion under OGD conditions. The time-course impedimetric sensing results showed that the proposed MEAs-based platform incorporated with a microperfusion environment can be used for the real-time monitoring of cortical neurons under in vitro OGD and the IGF-1 protective effect against OGD-induced ischemic neuronal death.
For AD disease model, we designed the disease propagation through a simple in vitro cortical neuronal network which were used for investigating the structural and functional differences between diseased and healthy neuronal populations at an axon-connected manner. The structural data via hyper-phosphorylated tau (hp-tau) and cytoskeleton alternation were captured from fluorescence imaging. Also, the functional data via spike number ratio and burst number per minute ratio were calculated from extracellular recording. From the structural data, we observed a disease propagation from diseased neuron population (hp-tau induced by okadaic-acid (OA) treatment), spreading through an axon-connected region then affecting the healthy neuron. The delayed response of neuritic degeneration from diseased neuron population propagated to the healthy neuron population was also observed through the neuronal cytoskeleton alternation from immunocytochemistry staining. From the functionality of AD cell models, we observed immediate electrophysiological pattern changes (spike number decreased) prior to the structural alternation occurring (neuritic degeneration). With advance in this simulated disease propagation model, the candidate treatment method or possible target mechanism could be validated.
In summary, this proposed MEAs-based platform provides electrophysiological characterization functionalities for in vitro/in vivo 3D neuronal network and also with integrated pathological environmental setup opens the way for specific neurological application. Our designed physiological/ pathological neuronal culture model not only could help the investigation of fundamental electrophysiological properties of 3D neuronal network but also reveal the potential treatment mechanism through neurological disease simulation environment.

Contents
致謝 IV
中文摘要 V
Abstract VII
Contents X
List of Tables ...XIIII
List of Figures ...XIIII

Chapter 1 Introduction
1.1 Introduction to planar microelectrode arrays 1
1.2 Impedance measurement of the neuron-electrode interface 2
1.3 Extracellular stimulation and recording of in vitro neuronal networks 3
1.4 Three-dimensional cell culture 4
1.5 Three-dimensional microelectrode arrays 6
1.6 Impedance measurement for in vitro 3D cellular environment 7
1.7 Extracellular recording of in vitro 3D neuronal networks 8
1.8 Novel flexible microelectrode arrays 9
1.9 Toward disease model application: in vitro ischemic stroke model 10
1.10 Impedimetric monitoring for in vitro ischemic stroke conditions 11
1.11 Toward disease model application: in vitro Alzheimer’s disease model 12
1.12 Hyper-phosphorylated microtubule-associate protein tau in Alzheimer’s disease 12
1.13 Compartmentalized microfluidic neuronal culture 13
1.14 The aims of this study 14

Chapter 2 Material and Methods
2.1 Fabrication and characterization of 3D tip-based MEAs 17
2.1.1 Fabrication process of 3D tip-based MEAs 17
2.1.2 In vitro 3D cortical neuronal culture 18
2.1.3 Florescence imaging of in vitro 3D cortical culture 19
2.1.4 Electrophysiological characterization of in vitro 3D neuronal networks 19

2.2 Fabrication and characterization of SU-8 Flexible Neuroprobe 20
2.2.1 Design and fabrication of SU-8 neuroprobe 20
2.2.2 Evaluation of mechanical properties of SU-8 neuroprobe 22
2.2.3 In vitro viability test and extracellular recording using neuroprobe 24
2.2.4 In vivo electrophysiological recording and immunobiological assays 24

2.3 Impedimetric monitoring of IGF-1 protection of in vitro cortical neurons under
ischemic conditions 27
2.3.1 Fabrication of microelectrode arrays 27
2.3.2 Cell culture and oxygen-glucose deprivation model 28
2.3.3 Impedance sensing under normal and OGD conditions 30
2.3.4 Measurement of cell viability and mitochondrial membrane potential 32
2.3.5 Statistical analysis 33

2.4 Disease Propagation of Okadaic Acid-Induced Tauopathy Model via
Compartmentalized MEAs Device 33
2.4.1 Design and integration of compartmentalized microfluidic device with
MEAs 33
2.4.2 Preparation and plating procedure for cortical neuronal culture 36
2.4.3 Induction of tau-hyperphosphorylation via okadaic acid treatment 37
2.4.4 Extracellular recording of cortical neuronal population inside
compartmentalized MEAs 38
2.4.5 Immunocytochemistry staining 39
2.4.6 Statistical analysis 40

Chapter 3 Results
3.1 Fabrication and characterization of 3D tip-based MEAs 41
3.1.1 Fabrication of 3D tip MEAs 41
3.1.2 Morphological observations and cell viability of 2D versus 3D cell culture 41
3.1.3 Impedimetric sensing of in vitro 3D neuronal network formation 43
3.1.4 Extracellular recording of in vitro 3D neuronal activities 45
3.2 Fabrication and characterization of SU-8 Flexible Neuroprobe 46
3.2.1 Fabrication and electrical characterization of SU-8 neuroprobe 46
3.2.2 Mechanical characterization of SU-8 neuroprobe 48
3.2.3 In vitro cytotoxicity testing assay and extracellular physiological recording 50
3.2.4 In vivo immune response and electrophysiological recording 52
3.3 Impedimetric monitoring of IGF-1 protection of in vitro cortical neurons under
ischemic conditions 54
3.3.1 Time-course resistive component of impedance monitoring of cell growth
on the electrode 54
3.3.2 Cell viability assessment 56
3.3.3 MMP assay 57
3.3.4 Time-course resistive component impedance monitoring of OGD-induced
cell detachment 59
3.4 Disease Propagation of Okadaic Acid-Induced Tauopathy Model via
Compartmentalized MEAs Device 60
3.4.1 Compartmentalized microfluidic device integrated with MEAs 60
3.4.2 Cytoskeleton alternation under OA-induced disease propagation 61
3.4.3 Propagation of tau hyper-phosphorylation between diseased and healthy
Neuronal populations 64
3.4.4 Spike number ratio of spontaneous activities during disease propagation 67
3.4.5 Bursts per minute ratio of spontaneous activities during disease propagation 68

Chapter 4 Discussion
4.1 Fabrication and characterization of 3D tip-based MEAs 71
4.2 Fabrication and characterization of SU-8 Flexible Neuroprobe 73
4.3 Impedimetric monitoring of IGF-1 protection of in vitro cortical neurons under
ischemic conditions 76
4.4 Disease Propagation of Okadaic Acid-Induced Tauopathy Model via
Compartmentalized MEAs Device 80

Chapter 5 Conclusion 88

References 91



List of Tables
Table 1 Summary of numerical data of spike number ratio and bursts per minute ratio under high and low concentration (600 nM and 60 nM, respectively) of OA treatment at all selected experiment time period 70

List of Figures
Figure.1 (a) Fabrication process of 3D tip MEAs. (b) Scanning electron microscopic imaging of tip structure. (c) Assembled 3D tip MEAs with external PCB for interfacing extracellular recording hardware. 18
Figure 2 Schematic representation of the fabrication processes for SU-8 neuroprobe in oblique and cross section views. (a) E-beam evaporation of Al sacrificial layer on a glass substrate, (b) spin-coating and definition of SU-8 neuroprobe pattern, (c) E-beam evaporation of Ti/Au electrode layer, (d) spin-coating and definition of electrode trace and pad by photoresist (PR), (e) etching of unwanted metal using wet etching solution, (f) removal of photoresist via acetone wash, (g) spin-coating and definition of opening via SU-8 patterning, (h) release of complete SU-8 neuroprobe via etching of Al sacrificial layer, and (i) layout of SU-8 neuroprobe with two groups of microelectrode for two different depths of brain regions, i.e. cortex and striatum.... 22
Figure 3 Mechanical analytical model of SU-8 neuroprobe during insertion process 23
Figure 4 Schematic diagrams of the fabrication of MEAs. (a) A 15-µm-thick photosensitive PI film was utilized as the substrate for the deposition of metals. (b) A gold layer (2000 Ǻ) was then deposited after a titanium layer (200 Ǻ) as the adhesion layer above the PI. Mask 1 was used to create the trace of electrodes by wet etching. (c) A gold electrode and pad trace pattern was formed by etched away unwanted metal then followed by removed the photoresist. (d) 20-µm-thick spin-coated PI film was used to encapsulate the electrodes. Mask 2 was then applied to expose microelectrodes and contact pads. (e) MEAs were then baked at 100 °C for 1 hr. The fabricated MEAs has total 60 microelectrodes and each active electrode site has diameter of 50 μm in circle. 28
Figure 5 Schematic diagram of microperfusion OGD system with electrophysiological measurement functionality. Whole measurement system could be easily equipped into inverted microscope for real-time observation of cell behavior. 30
Figure 6 Photos of (a) integrated devices consisting of compartmentalized microfluidic device and microelectrode array; (b) microscopic view of three culture region with twenty microelectrode arrangement; (c) fluorescence imaging of separated neuronal population connected with axons (green: MAP II, red: Smi 312, blue: DAPI). .. 36
Figure 7 Phase-contrast microscopic imaging for observing of morphological difference between 2D and 3D neuronal culture: (a) 2D neuronal network in petri-dish surface, (b) 2D neuronal culture in 3D tip MEAs surface, (c) Matrigel-based 3D neuronal culture inside petri-dish culture, (d) Matrigel –based 3D neuronal culture inside 3D tip MEAs culture.. 42
Figure 8 Cell viability of 2D and 3D neuronal culture during 21 DIVs culture period. The observation time point was selected as following: after 4 hours cell seeding as 0 DIVs, 7 DIVS, 14 DIVs and 21 DIVs, (n=10, *p〈0.05). The cell viability was calculated as the number of live cells divided by the sum of live and dead cells 43
Figure 9 Impedimetric measurement results during 2D and 3D neuronal network formation during 21 DIVs culture period..... 44
Figure 10 Typical extracellular recorded spike from in vitro 3D neuronal network after 17 DIVs culture period 45
Figure 11 Spike raster train from in vitro 3D neuronal network after 17 DIVs culture period. 46
Figure 12 SEM images of SU-8 neuroprobe of (a) whole probe structure and (b) one group of microelectrodes with four microelectrodes for electrophysiological recording (circular shape) and one reserved for future integration of electrochemical sensing (rectangular shape). Higher magnification of (c) single recording microelectrode and (d) wire bonding to external PCB for recordings... 47
Figure 13 Mechanical evaluation of SU-8 neuroprobe during insertion process and images acquired during (a) buckling test and (b) bending test... 49
Figure 14 Insertion and retraction behavior of flexible SU-8 neuroprobe during three different samples. The insertion resistance force exhibited different patterns for 0.6% agar brain phantom (solid line), brain with dura and pia (dotted line), and brain with pia only (dashed line). The insertion of neuroprobe was operated at fixed speed of 100 µm/s to around 6 mm in depth. Before retraction at 80 sec, the resistance force dropped to about zero after insertion.... 49
Figure 15 Cytoxicity test of SU-8 neuroprobe. Cortical neurons cultured in culture dish for (a) 7 DIVs and (c) 14 DIVs. Cortical neurons cultured on SU-8 neuroprobe for (b) 7 DIVs and (d) 14 DIVs. (e) Quantification of cell viability for culture and neuroprobe (N=10 for each group).... 51
Figure 16 Extracellular spontaneous action potential of cortical neurons cultured on SU-8 neuroprobe was recorded from 17-DIV cortical neurons, (a). Magnified view of signals recorded from SU-8 neuroprobe can be distinguished from background noise, (b).... 52
Figure 17 Immunohistological staining of tissue reaction around implantation site. Anti-MAPII/GFAP staining around implanted neuroprobe in (a) cortex region and (b) striatum 14 days after implantation (anti-MAPII stained red for neuron marker; anti-GFAP stained green for astrocyte marker).... 53
Figure 18 (a) Electrophysiological recording of neural signals after SU-8 neuroprobe implantation. Two action potential signals extracted from those marked with arrows in (a)with (b) minimum and (c) maximum SNR.... 54
Figure 19 Time-course monitoring results of impedance of cells on the electrode over 7 DIVs (n=4). The curve of cortical neuron represents neurons cultured for 7 DIVs. The cells were deliberately damaged on day 8 via adjustment of osmolarity.... 55
Figure 20 Cell viabilities of control and IGF-1-protected (50 ng/mL and 100 ng/mL) group of rat cortical neurons before OGD, after 30-min of OGD, and after 24 hr of re-oxygenation (re-oxy), (n=4, *p 〈 0.05). The cell viability was calculated as the number of live cells divided by the sum of live and dead cells.... 57
Figure 21 Normalized JC-1 fluorescence change of normal and IGF-1-protected (50 ng/mL and 100 ng/mL) cortical neurons during 30-min of OGD and 2 hr of re-oxygenation. JC-1 fluorescence was normalized and valinomycin was used to force the MMP to be at the level of depolarization.... 58
Figure 22 Impedimetric monitoring results of normal and IGF-1-protected rat cortical neurons during 30-min of OGD and controls without OGD insult.... 60
Figure 23 Pictures of (a) microscopic view of three culture region with twenty microelectrode arrangement, (b) fluorescence imaging of separated neuronal population connected with axon (green: MAP II, red: Smi 312, blue: DAPI).... 61
Figure 24 The diseased and healthy neuronal population under OA and without treatment. The cytoskeleton alternation visualized as green color by MAP II immunocytochemistry staining for (a) high concentration OA treatment (600 nM), (b) low concentration OA treatment (60 nM).,,.... 63
Figure 25 Disease propagation process during high concentration OA treatment (600 nM). Hp-tau was visualized as green color by pS262 immunocytochemistry staining..... 66
Figure 26 Disease propagation process during low concentration OA treatment (60 nM). Hp-tau was visualized as green color by pS262 immunocytochemistry staining.... 66
Figure 27 Spike number ratio acquired by extracellular recording during two concentration of OA treatment (600 nM, 60 nM) which represented the functional pattern changed during disease propagation, (N = 5, * p 〈 0.05, ** p 〈 0.01, *** p 〈 0.001, **** p 〈 0.005), The spike number ratio was calculated as the value of each recovery period (75min + 0h, 6h, 24h and 48h) divided by the value of before 75min OA as control value..... 68
Figure 28 Bursts per minute ratio acquired by extracellular recording during two concentration of OA treatment (600 nM, 60 nM) which represented the functional pattern changed during disease propagation, (N = 5, * p 〈 0.05, ** p 〈 0.01, *** p 〈 0.005, **** p 〈 0.001), The bursts per minute ratio was calculated as the value of each recovery period (75min + 0h, 6h, 24h and 48h) divided by the value of before 75min OA as control value.... 70

References
V. Aas, S. Torbla, M. H. Andersen, J. Jensen, A. C. Rustan, Electrical stimulation improves insulin responses in a human skeletal muscle cell model of hyperglycemia, Annals of the New York Academy of Sciences, vol. 967, pp. 506-515, 2002.
P. Abgrall, V. Conedera, H. Camon, A. a. Gue, and N. r. Nguyen, SU-8 as a structural material for labs on chips and microelectromechanical systems, Electrophoresis, vol. 28, pp. 4539-4551, 2007.
A. d. C. Alonso, I. Grundke-Iqbal, H. S. Barra, and K. Iqbal, Abnormal phosphorylation of tau and the mechanism of Alzheimer neurofibrillary degeneration: sequestration of microtubule-associated proteins 1 and 2 and the disassembly of microtubules by the abnormal tau, Proceedings of the National Academy of Sciences, vol. 94, pp. 298-303, 1997.
A. Altuna, G. Gabriel, L. M. de la Prida, M. Tijero, A. Guimera, J. Berganzo, R. Salido, R. Villa, and L. J. Fernandez, SU-8-based microneedles for in vitro neural applications, Journal of Micromechanics and Microengineering, vol. 20, p. 064014, 2010.
A. Altuna, L. Menendez de la Prida, E. Bellistri, G. Gabriel, A. Guimera, J. Berganzo, R. Villa, and L. J. Fernandez, SU-8 based microprobes with integrated planar electrodes for enhanced neural depth recording, Biosensors and Bioelectronics, vol. 37, pp. 1-5, 2012.
P. Aivar, M. Valero, E. Bellistri, and L. M. de la Prida, Extracellular Calcium Controls the Expression of Two Different Forms of Ripple-Like Hippocampal Oscillations, The Journal of Neuroscience, vol. 34, pp. 2989-3004, 2014.
J. F. Alvarez-Barreto, M. C. Shreve, P. L. Deangelis, and V. I. Sikavitsas, Preparation of a functionally flexible, three-dimensional, biomimetic poly (L-lactic acid) scaffold with improved cell adhesion, Tissue engineering, vol. 13, pp. 1205-1217, 2007.
H. Andersson and A. Van Den Berg, Microfabrication and microfluidics for tissue engineering: state of the art and future opportunities, Lab Chip, vol. 4, pp. 98-103, 2004.
H. Arai, M. Terajima, M. Miura, S. Higuchi, T. Muramatsu, N. Machida, H. Seiki, S. Takase, C. M. Clark, and V. M. Lee, Tau in cerebrospinal fluid: a potential diagnostic marker in Alzheimer's disease, Annals of neurology, vol. 38, pp. 649-652, 1995.
C. Arias, N. Sharma, P. Davies, and B. Shafit-zagardo, Okadaic Acid Induces Early Changes in Microtubule-associated Protein 2 and Tau Phosphorylation Prior to Neurodegeneration in Cultured Cortical Neurons, Journal of neurochemistry, vol. 61, pp. 673-682, 1993.
S. Arndt, J. Seebach, K. Psathaki, H.-J. Galla, and J. Wegener, Bioelectrical impedance assay to monitor changes in cell shape during apoptosis, Biosensors and Bioelectronics, vol. 19, pp. 583-594, 2004.
F. Asphahani and M. Zhang, Cellular impedance biosensors for drug screening and toxin detection, Analyst, vol. 132, pp. 835-841, 2007.
F. Asphahani, M. Thein, O. Veiseh, D. Edmondson, R. Kosai, M. Veiseh, J. Xu, and M. Zhang, Influence of cell adhesion and spreading on impedance characteristics of cell-based sensors, Biosensors and Bioelectronics, vol. 23, pp. 1307-1313, 2008.
T. Atkinson, J. Whitfield, and B. Chakravarthy, The phosphatase inhibitor, okadaic acid, strongly protects primary rat cortical neurons from lethal oxygen?lucose deprivation, Biochemical and biophysical research communications, vol. 378, pp. 394-398, 2009.
C. Ballatore, V. M. Y. Lee, and J. Q. Trojanowski, Tau-mediated neurodegeneration in Alzheimer's disease and related disorders, Nature Reviews Neuroscience, vol. 8, pp. 663-672, 2007.
R. E. Becker, N. H. Greig, and E. Giacobini, Why do so many drugs for Alzheimer's disease fail in development? Time for new methods and new practices?, Journal of Alzheimer's Disease, vol. 15, pp. 303-325, 2008.
R. E. Becker and N. H. Greig, Why so few drugs for Alzheimer's disease? Are methods failing drugs?, Current Alzheimer Research, vol. 7, p. 642, 2010.
R. E. Becker and N. H. Greig, Increasing the success rate for Alzheimer's disease drug discovery and development, Expert opinion on drug discovery, vol. 7, pp. 367-370, 2012
R. Biran, D. C. Martin, and P. A. Tresco, Neuronal cell loss accompanies the brain tissue response to chronically implanted silicon microelectrode arrays, Experimental neurology, vol. 195, pp. 115-126, 2005.
R. Brandt, M. Hundelt, and N. Shahani, Tau alteration and neuronal degeneration in tauopathies: mechanisms and models, Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease, vol. 1739, pp. 331-354, 2005.
G. J. Brewer, M. D. Boehler, S. Leondopulos, L. Pan, S. Alagapan, T. B. DeMarse, and B. C. Wheeler, Toward a self-wired active reconstruction of the hippocampal trisynaptic loop: DG-CA3, Frontiers in neural circuits, vol. 7, 2013.
J. M. Boyd, L. Huang, L. Xie, B. Moe, S. Gabos, and X.-F. Li, A cell-microelectronic sensing technique for profiling cytotoxicity of chemicals, Analytica chimica acta, vol. 615, pp. 80-87, 2008.
J. R. Buitenweg, W. L. C. Rutten, E. Marani, S. K. L. Polman, and J. Ursum, Extracellular detection of active membrane currents in the neuron-electrode interface, Journal of neuroscience methods, vol. 115, pp. 211-221, 2002.
A. de Calignon, M. Polydoro, M. Suarez-Calvet, C. William, D. H. Adamowicz, K. J. Kopeikina, R. Pitstick, N. Sahara, K. H. Ashe, and G. A. Carlson, Propagation of tau pathology in a model of early Alzheimer's disease, Neuron, vol. 73, pp. 685-697, 2012.
D. C. Chen, J. R. Avansino, V. G. Agopian, V. D. Hoagland, J. D. Woolman, S. Pan, B. D. Ratner, and M. Stelzner, Comparison of polyester scaffolds for bioengineered intestinal mucosa, Cells Tissues Organs, vol. 184, pp. 154-165, 2007.
K. Cheung, Y. Zhong, P. Renaud, and R. Bellamkonda, Comparison of tissue reaction to implanted polyimide and silicon microelectrode arrays, European Cells and Materials, vol. 10, p. 5, 2005.
K. C. Cheung, P. Renaud, H. Tanila, and K. Djupsund, Flexible polyimide microelectrode array for in vivo recordings and current source density analysis, Biosensors and Bioelectronics, vol. 22, pp. 1783-1790, 2007.
Y.-Y. Chen, H.-Y. Lai, S.-H. Lin, C.-W. Cho, W.-H. Chao, C.-H. Liao, S. Tsang, Y.-F. Chen, and S.-Y. Lin, Design and fabrication of a polyimide-based microelectrode array: Application in neural recording and repeatable electrolytic lesion in rat brain, Journal of neuroscience methods, vol. 182, pp. 6-16, 2009.
M. Chiappalone, M. Bove, A. Vato, M. Tedesco, and S. Martinoia, Dissociated cortical networks show spontaneously correlated activity patterns during in vitro development, Brain research, vol. 1093, pp. 41-53, 2006.
F. Clavaguera, T. Bolmont, R. A. Crowther, D. Abramowski, S. Frank, A. Probst, G. Fraser, A. K. Stalder, M. Beibel, and M. Staufenbiel, Transmission and spreading of tauopathy in transgenic mouse brain, Nature cell biology, vol. 11, pp. 909-913, 2009.
Cho, S. H., H. M. Lu., Cauller, L., Romero-Ortega, M.I., Jeong-Bong Lee., and Hughes, G.A., Biocompatible SU-8-based microprobes for recording neural spike signals from regenerated peripheral nerve fibers, Sensors Journal, vol. 8, pp. 1830-1836, 2008.
P. W. Coates and R. D. Nathan, Feasibility of electrical recordings from unconnected vertebrate CNS neurons cultured in a three-dimensional extracellular matrix, Journal of neuroscience methods, vol. 20, pp. 203-210, 1987.
J. L. Crimins, A. B. Rocher, A. Peters, P. Shultz, J. Lewis, and J. I. Luebke, Homeostatic responses by surviving cortical pyramidal cells in neurodegenerative tauopathy, Acta neuropathologica, vol. 122, pp. 551-564, 2011.
J. L. Crimins, A. B. Rocher, and J. I. Luebke, Electrophysiological changes precede morphological changes to frontal cortical pyramidal neurons in the rTg4510 mouse model of progressive tauopathy, Acta neuropathologica, vol. 124, pp. 777-795, 2012.
X. Delbeuck, M. Van der Linden, and F. Collette, Alzheimer'Disease as a Disconnection Syndrome?, Neuropsychology review, vol. 13, pp. 79-92, 2003.
Discher, D. E., P. Janmey, and Y Wang, Tissue cells feel and respond to the stiffness of their substrate, Science, vol. 310, pp. 5751, 2005.
R. Djakaria, B. I. Chandran, M. H. Gordon, W. F. Schmidt, and T. G. Lenihan, Determination of Young's modulus of thin films used in embedded passive devices, in Electronic Components and Technology Conference, 1997. Proceedings., 47th, pp. 745-749, 1997.
B. J. Dworak and B. C. Wheeler, Novel MEA platform with PDMS microtunnels enables the detection of action potential propagation from isolated axons in culture, Lab on a Chip, vol. 9, pp. 404-410, 2009.
J. El-Ali, P. K. Sorger, and K. F. Jensen, Cells on chips, Nature, vol. 442, pp. 403-411, 2006.
J. P. Frampton, M. R. Hynd, J. C. Williams, M. L. Shuler, and W. Shain, Three-dimensional hydrogel cultures for modeling changes in tissue impedance around microfabricated neural probes, Journal of neural engineering, vol. 4, p. 399, 2007.
J. M. Goddard and J. H. Hotchkiss, Polymer surface modification for the attachment of bioactive compounds, Progress in polymer science, vol. 32, pp. 698-725, 2007.
P. Gluckman, N. Klempt, J. Guan, C. Mallard, E. Sirimanne, M. Dragunow, M. Klempt, K. Singh, C. Williams, and K. Nikolics, A role for IGF-1 in the rescue of CNS neurons following hypoxic-ischemic injury, Biochemical and biophysical research communications, vol. 182, pp. 593-599, 1992.
S. Gerecht-Nir, S. Cohen, A. Ziskind, and J. Itskovitz-Eldor, Three-dimensional porous alginate scaffolds provide a conducive environment for generation of well-vascularized embryoid bodies from human embryonic stem cells, Biotechnology and bioengineering, vol. 88, pp. 313-320, 2004.
M. P. Goldberg and D. W. Choi, Combined oxygen and glucose deprivation in cortical cell culture: calcium-dependent and calcium-independent mechanisms of neuronal injury, The Journal of neuroscience, vol. 13, pp. 3510-3524, 1993.
M. Guo, J. Chen, X. Yun, K. Chen, L. Nie, and S. Yao, Monitoring of cell growth and assessment of cytotoxicity using electrochemical impedance spectroscopy, Biochimica et Biophysica Acta (BBA)-General Subjects, vol. 1760, pp. 432-439, 2006.
L. Haines-Butterick, K. Rajagopal, M. Branco, D. Salick, R. Rughani, M. Pilarz, M. S. Lamm, D. J. Pochan, and J. P. Schneider, Controlling hydrogelation kinetics by peptide design for three-dimensional encapsulation and injectable delivery of cells, Proceedings of the National Academy of Sciences, vol. 104, pp. 7791-7796, 2007.
N. Haj Hosseini, R. Hoffmann, S. Kisban, T. Stieglitz, O. Paul, and P. Ruther, Comparative study on the insertion behavior of cerebral microprobes, in Engineering in Medicine and Biology Society, 2007. EMBS 2007. 29th Annual International Conference of the IEEE, pp. 4711-4714, 2007.
M. Hajj-Hassan, S. Musallam, and V. Chodavarapu, Reinforced silicon neural microelectrode array fabricated using a commercial MEMS process, Journal of Micro/Nanolithography, MEMS, and MOEMS, vol. 8, pp. 033011-033011-8, 2009.
T. J. Hall, M. Bilgen, M. F. Insana, and T. A. Krouskop, Phantom materials for elastography, Ultrasonics, Ferroelectrics and Frequency Control, IEEE Transactions on, vol. 44, pp. 1355-1365, 1997.
J. A. Hardy and G. A. Higgins, Alzheimer's disease: the amyloid cascade hypothesis, Science, 1992.
J. Hardy and D. J. Selkoe, The amyloid hypothesis of Alzheimer's disease: progress and problems on the road to therapeutics, Science, vol. 297, pp. 353-356, 2002.
J. Hardy, The amyloid hypothesis for Alzheimer‘s disease: a critical reappraisal, Journal of neurochemistry, vol. 110, pp. 1129-1134, 2009.
J. A. Harris, N. Devidze, L. Verret, K. Ho, B. Halabisky, M. T. Thwin, D. Kim, P. Hamto, I. Lo, and G.-Q. Yu, Transsynaptic progression of amyloid-β-induced neuronal dysfunction within the entorhinal-hippocampal network, Neuron, vol. 68, pp. 428-441, 2010.
J. P. Harris, A. E. Hess, S. J. Rowan, C. Weder, C. A. Zorman, D. J. Tyler, and J. R. Capadona, In vivo deployment of mechanically adaptive nanocomposites for intracortical microelectrodes, Journal of neural engineering, vol. 8, p. 046010, 2011.
C. Hassler, T. Boretius, and T. Stieglitz, Polymers for neural implants, Journal of Polymer Science Part B: Polymer Physics, vol. 49, pp. 18-33, 2011.
Y. He, Z. Chen, G. Gong, and A. Evans, Neuronal networks in Alzheimer's disease, The Neuroscientist, vol. 15, pp. 333-350, 2009.
A. Heller, H. Choi, and L. Won, Regulation of developing dopaminergic axonal arbor size in three-dimensional reaggregate tissue culture, Journal of Comparative Neurology, vol. 384, pp. 349-358, 1997.
M. O. Heuschkel, M. Fejtl, M. Raggenbass, D. Bertrand, and P. Renaud, A three-dimensional multi-electrode array for multi-site stimulation and recording in acute brain slices, Journal of neuroscience methods, vol. 114, pp. 135-148, 2002.
I. T. Hsieh, C. C.-H. Yang, C.-Y. Chen, G.-S. Lee, F.-J. Kao, K.-L. Kuo, and T. B.-J. Kuo, Uninterrupted wireless long-term recording of sleep patterns and autonomic function in freely moving rats, Journal of Medical and Biological Engineering, vol. 33, pp. 79-86, 2013.
S.-H. Huang, S.-P. Lin, C.-K. Liang, and J.-J. J. Chen, Impedimetric monitoring of IGF-1 protection of in vitro cortical neurons under ischemic conditions, Biomedical microdevices, vol. 15, pp. 135-143, 2013.
T. S. Hug, Biophysical methods for monitoring cell-substrate interactions in drug discovery, Assay and drug development technologies, vol. 1, pp. 479-488, 2003.
T. Iijima, Mitochondrial membrane potential and ischemic neuronal death, Neuroscience research, vol. 55, pp. 234-243, 2006.
K. Iqbal, A. d. C. Alonso, and I. Grundke-Iqbal, Cytosolic abnormally hyperphosphorylated tau but not paired helical filaments sequester normal MAPs and inhibit microtubule assembly, Journal of Alzheimer's Disease, vol. 14, pp. 365-370, 2008.
H. R. Irons, D. K. Cullen, N. P. Shapiro, N. A. Lambert, R. H. Lee, and M. C. LaPlaca, Three-dimensional neural constructs: a novel platform for neurophysiological investigation, Journal of neural engineering, vol. 5, p. 333, 2008.
T. Ishihara, M. Hong, B. Zhang, Y. Nakagawa, M. K. Lee, J. Q. Trojanowski, and V. M. Y. Lee, Neuron, 24,751-762, 1999.
L. M. Ittner and J. Gotz, Amyloid-β and tau - toxic pas de deux in Alzheimer's disease, Nature Reviews Neuroscience, vol. 12, pp. 67-72, 2010.
T. Jacobs, T. Valero, M. Naumann, S. Kintzios, and P. Hauptmann, Electrical impedance spectroscopy of gel embedded neuronal cells based on a novel impedimetric biosensor, Procedia Chemistry, vol. 1, pp. 261-264, 2009.
D. C. Jean and P. W. Baas, It cuts two ways: microtubule loss during Alzheimer disease, The EMBO journal, vol. 32, pp. 2900-2902, 2013.
I. Jin, E. R. Kandel, and R. D. Hawkins, Whereas short-term facilitation is presynaptic, intermediate-term facilitation involves both presynaptic and postsynaptic protein kinases and protein synthesis, Learning ＆ Memory, vol. 18, pp. 96-102, 2011.
M. D. Johnson, R. K. Franklin, M. D. Gibson, R. B. Brown, and D. R. Kipke, Implantable microelectrode arrays for simultaneous electrophysiological and neurochemical recordings, Journal of neuroscience methods, vol. 174, pp. 62-70, 2008.
A. F. M. Johnstone, G. W. Gross, D. G. Weiss, O. H. U. Schroeder, A. Gramowski, and T. J. Shafer, Microelectrode arrays: a physiologically based neurotoxicity testing platform for the 21st century, Neurotoxicology, vol. 31, pp. 331-350, 2010.
P. K. Kamat, S. Rai, and C. Nath, Okadaic acid induced neurotoxicity: An emerging tool to study Alzheimer's disease pathology, Neurotoxicology, vol. 37, pp. 163-172, 2013.
T. T. Kanagasabapathi, D. Ciliberti, S. Martinoia, W. J. Wadman, and M. M. J. Decre, Dual-compartment neurofluidic system for electrophysiological measurements in physically segregated and functionally connected neuronal cell culture, Frontiers in neuroengineering, vol. 4, 2011.
T. T. Kanagasabapathi, P. Massobrio, R. A. Barone, M. Tedesco, S. Martinoia, W. J. Wadman, and M. M. J. Decre, Functional connectivity and dynamics of cortical-thalamic networks co-cultured in a dual compartment device, Journal of neural engineering, vol. 9, p. 036010, 2012.
P. Massobrio, C. N. G. Giachello, M. Ghirardi, and S. Martinoia, Selective modulation of chemical and electrical synapses of Helix neuronal networks during in vitro development, BMC neuroscience, vol. 14, p. 22., 2013.
J. M. Kelm, N. E. Timmins, C. J. Brown, M. Fussenegger, and L. K. Nielsen, Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types, Biotechnology and Bioengineering, vol. 83, pp. 173-180, 2003.
E. G. R. Kim, J. K. John, H. Tu, Q. Zheng, J. Loeb, J. Zhang, and Y. Xu, A hybrid silicon-parylene neural probe with locally flexible regions, Sensors and Actuators B: Chemical, 2014.
W. Kim, S. Lee, C. Jung, A. Ahmed, G. Lee, and G. F. Hall, Interneuronal transfer of human tau between Lamprey central neurons in situ, Journal of Alzheimer's Disease, vol. 19, pp. 647-664, 2010.
H. K. Kleinman and G. R. Martin, Matrigel: basement membrane matrix with biological activity, in Seminars in cancer biology, pp. 378-386, 2005.
D. Klo, R. Kurz, H.-G. Jahnke, M. Fischer, A. Rothermel, U. Anderegg, J. C. Simon, and A. A. Robitzki, Microcavity array (MCA)-based biosensor chip for functional drug screening of 3D tissue models, Biosensors and Bioelectronics, vol. 23, pp. 1473-1480, 2008.
T. D. Y. Kozai and D. R. Kipke, Insertion shuttle with carboxyl terminated self-assembled monolayer coatings for implanting flexible polymer neural probes in the brain, Journal of neuroscience methods, vol. 184, pp. 199-205, 2009.
A. V. Kravitz and A. C. Kreitzer, Optogenetic manipulation of neural circuitry in vivo Current opinion in neurobiology, vol. 21, pp. 433-439, 2011.
D. Krinke, H.-G. Jahnke, O. Panke, and A. A. Robitzki, A microelectrode-based sensor for label-free in vitro detection of ischemic effects on cardiomyocytes, Biosensors and Bioelectronics, vol. 24, pp. 2798-2803, 2009.
I. I. Kruman and M. P. Mattson, Pivotal role of mitochondrial calcium uptake in neural cell apoptosis and necrosis, Journal of neurochemistry, vol. 72, pp. 529-540, 1999.
A. Kunze, R. Meissner, S. Brando, and P. Renaud, Co-pathological connected primary neurons in a microfluidic device for alzheimer studies, Biotechnology and bioengineering, vol. 108, pp. 2241-2245, 2011.
I. Kuperstein, K. Broersen, I. Benilova, J. Rozenski, W. Jonckheere, M. Debulpaep, A. Vandersteen, I. Segers-Nolten, K. Van Der Werf, and V. Subramaniam, Neurotoxicity of Alzheimer's disease Aβ peptides is induced by small changes in the Aβ42 to Aβ40 ratio, The EMBO journal, vol. 29, pp. 3408-3420, 2010.
J. Kuret, E. E. Congdon, G. Li, H. Yin, X. Yu, and Q. I. Zhong, Evaluating triggers and enhancers of tau fibrillization, Microscopy research and technique, vol. 67, pp. 141-155, 2005.
M. Kusumoto, E. Dux, W. Paschen, and K. Hossmann, Susceptibility of Hippocampal and Cortical Neurons to Argon-Mediated In vitro Ischemia, Journal of neurochemistry, vol. 67, pp. 1613-1621, 1996.
A. H. Kyle, C. T. O. Chan, and A. I. Minchinton, Characterization of three-dimensional tissue cultures using electrical impedance spectroscopy, Biophysical journal, vol. 76, pp. 2640-2648, 1999.
S. P. Lacour, S. Benmerah, E. Tarte, J. FitzGerald, J. Serra, S. McMahon, J. Fawcett, O. Graudejus, Z. Yu, and B. Morrison Iii, Flexible and stretchable micro-electrodes for in vitro and in vivo neural interfaces, Medical ＆ biological engineering ＆ computing, vol. 48, pp. 945-954, 2010.
J. B. Leach, A. K. H. Achyuta, and S. K. Murthy, Bridging the divide between neuroprosthetic design, tissue engineering and neurobiology, Frontiers in neuroengineering, vol. 2, 2009.
J. Lee, M. J. Cuddihy, and N. A. Kotov, “Three-dimensional cell culture matrices: state of the art, Tissue Engineering Part B: Reviews, vol. 14, no. 1, pp. 61-86, 2008.
K. Lee, A. Singh, J. He, S. Massia, B. Kim, and G. Raupp, Polyimide based neural implants with stiffness improvement, Sensors and Actuators B: Chemical, vol. 102, pp. 67-72, 2004.
K.-N. Lee, D.-S. Shin, Y.-S. Lee, and Y.-K. Kim, Micromirror array for protein micro array fabrication, Journal of Micromechanics and Microengineering, vol. 13, p. 474, 2003.
S. Lee, W. Kim, Z. Li, and G. F. Hall, Accumulation of vesicle-associated human tau in distal dendrites drives degeneration and tau secretion in an in situ cellular tauopathy model, International journal of Alzheimer's disease, vol. 2012.
K. Leroy, Z. Yilmaz, and J. Brion, Increased level of active GSK-3 in Alzheimer‘s disease and accumulation in argyrophilic grains and in neurones at different stages of neurofibrillary degeneration, Neuropathology and applied neurobiology, vol. 33, pp. 43-55, 2007.
J. Lewis, E. McGowan, J. Rockwood, H. Melrose, P. Nacharaju, M. Van Slegtenhorst, K. Gwinn-Hardy, M. P. Murphy, M. Baker, and X. Yu, Neurofibrillary tangles, amyotrophy and progressive motor disturbance in mice expressing mutant (P301L) tau protein, Nature genetics, vol. 25, pp. 402-405, 2000.
D. Lewitus, K. L. Smith, W. Shain, and J. Kohn, Ultrafast resorbing polymers for use as carriers for cortical neural probes, Acta biomaterialia, vol. 7, pp. 2483-2491.
D. Y. Lewitus, K. L. Smith, W. Shain, D. Bolikal, and J. Kohn, The fate of ultrafast degrading polymeric implants in the brain, Biomaterials, vol. 32, pp. 5543-5550, 2011.
S.-P. Lin, J.-J. J. Chen, J.-D. Liao, and S.-F. Tzeng, Characterization of surface modification on microelectrode arrays for in vitro cell culture, Biomedical microdevices, vol. 10, pp. 99-111, 2008.
S. P. Lin, T. R. Kyriakides, and J. J. J. Chen, “On-line observation of cell growth in a three-dimensional matrix on surface-modified microelectrode arrays, Biomaterials, vol. 30, no. 17, pp. 3110-3117, 2009.
W.-L. Lin, J. Lewis, S.-H. Yen, M. Hutton, and D. W. Dickson, Ultrastructural neuronal pathology in transgenic mice expressing mutant (P301L) human tau, Journal of neurocytology, vol. 32, pp. 1091-1105, 2003.
P. Linderholm, J. Vannod, Y. Barrandon, and P. Renaud, Bipolar resistivity profiling of 3D tissue culture, Biosensors and Bioelectronics, vol. 22, pp. 789-796, 2007.
Q. Liu, J. Yu, L. Xiao, J. C. O. Tang, Y. Zhang, P. Wang, M. Yang, Biosensors and Bioelectronic, vol. 24, pp. 1305-1310, 2009.
L. Liu, V. Drouet, J. W. Wu, M. P. Witter, S. A. Small, C. Clelland, and K. Duff, Trans-synaptic spread of tau pathology in vivo, PloS one, vol. 7, p. e31302, 2012.
H. Lorenz, M. Despont, N. Fahrni, N. LaBianca, P. Renaud, and P. Vettiger, SU-8: a low-cost negative resist for MEMS, Journal of Micromechanics and Microengineering, vol. 7, p. 121, 1997.
J. C. Lotters, W. Olthuis, P. H. Veltink, and P. Bergveld, The mechanical properties of the rubber elastic polymer polydimethylsiloxane for sensor applications, Journal of Micromechanics and Microengineering, vol. 7, p. 145, 1997.
A. E. Ludvigson, J. I. Luebke, J. Lewis, and A. Peters, Structural abnormalities in the cortex of the rTg4510 mouse model of tauopathy: a light and electron microscopy study, Brain Structure and Function, vol. 216, pp. 31-42, 2011.
J. H. T. Luong, M. Habibi-Rezaei, J. Meghrous, C. Xiao, K. B. Male, and A. Kamen, Monitoring motility, spreading, and mortality of adherent insect cells using an impedance sensor, Analytical chemistry, vol. 73, pp. 1844-1848, 2001.
M. J. Mahoney and K. S. Anseth, Three-dimensional growth and function of neural tissue in degradable polyethylene glycol hydrogels, Biomaterials, vol. 27, pp. 2265-2274, 2006.
C. Marin and E. Fernandez, Biocompatibility of intracortical microelectrodes: current status and future prospects, Frontiers in neuroengineering, vol. 3, p. 8, 2010.
M. Martinez-Sanchez, F. Striggow, U. H. Schroder, S. Kahlert, K. G. Reymann, and G. Reiser, Na2+ and Ca2+ homeostasis pathways, cell death and protection after oxygen-glucose-deprivation in organotypic hippocampal slice cultures, Neuroscience, vol. 128, pp. 729-740, 2004.
S. Martinoia, L. Bonzano, M. Chiappalone, M. Tedesco, M. Marcoli, and G. Maura, In vitro cortical neuronal networks as a new high-sensitive system for biosensing applications, Biosensors and Bioelectronics, vol. 20, pp. 2071-2078, 2005.
M. P. Mazanetz and P. M. Fischer, Untangling tau hyperphosphorylation in drug design for neurodegenerative diseases, Nature reviews Drug discovery, vol. 6, pp. 464-479, 2007.
R. Meissner, B. Eker, H. Kasi, A. Bertsch, and P. Renaud, Distinguishing drug-induced minor morphological changes from major cellular damage via label-free impedimetric toxicity screening, Lab on a chip, vol. 11, pp. 2352-2361, 2011.
A. Mercanzini, K. Cheung, D. L. Buhl, M. Boers, A. Maillard, P. Colin, J.-C. Bensadoun, A. Bertsch, and P. Renaud, Demonstration of cortical recording using novel flexible polymer neural probes, Sensors and Actuators A: Physical, vol. 143, pp. 90-96, 2008.
F. O. Morin, Y. Takamura, and E. Tamiya, “Investigating neuronal activity with planar microelectrode arrays: achievements and new perspectives, Journal of bioscience and bioengineering, vol. 100, no. 2, pp. 131-143, 2005.
G. Morfini, G. Szebenyi, r. ouml, B. Richards, and S. T. Brady, Regulation of kinesin: implications for neuronal development, Developmental neuroscience, vol. 23, pp. 364-376, 2001.
G. A. Morfini, M. Burns, L. I. Binder, N. M. Kanaan, N. LaPointe, D. A. Bosco, R. H. Brown, H. Brown, A. Tiwari, and L. Hayward, Axonal transport defects in neurodegenerative diseases, The Journal of Neuroscience, vol. 29, pp. 12776-12786, 2009.
K. Najafi and J. F. Hetke, Strength characterization of silicon microprobes in neurophysiological tissues, Biomedical Engineering, IEEE Transactions on, vol. 37, pp. 474-481, 1990.
H.-s. Noh, K.-s. Moon, A. Cannon, P. J. Hesketh, and C. P. Wong, Wafer bonding using microwave heating of parylene intermediate layers, Journal of Micromechanics and Microengineering, vol. 14, p. 625, 2004.
S.-M. Ong, C. Zhang, Y.-C. Toh, S. H. Kim, H. L. Foo, C. H. Tan, D. van Noort, S. Park, and H. Yu, A gel-free 3D microfluidic cell culture system, Biomaterials, vol. 29, pp. 3237-3244, 2008
T. J. O'Shaughnessy, H. J. Lin, and W. Ma, “Functional synapse formation among rat cortical neurons grown on three-dimensional collagen gels, Neuroscience letters, vol. 340, no. 3, pp. 169-172, 2003.
J. J. Palop, J. Chin, and L. Mucke, A network dysfunction perspective on neurodegenerative diseases, Nature, vol. 443, pp. 768-773, 2006.
J. J. Palop and L. Mucke, Amyloid-[beta]-induced neuronal dysfunction in Alzheimer's disease: from synapses toward neural networks, Nature neuroscience, vol. 13, pp. 812-818, 2010.
F. Pampaloni, E. G. Reynaud, and E. H. K. Stelzer, “The third dimension bridges the gap between cell culture and live tissue, Nature Reviews Molecular Cell Biology, vol. 8, no. 10, pp. 839-845, 2007.
D. Puzzo, L. Privitera, E. Leznik, M. F, A. Staniszewski, A. Palmeri, and O. Arancio, Picomolar amyloid-β positively modulates synaptic plasticity and memory in hippocampus, The Journal of Neuroscience, vol. 28, pp. 14537-14545, 2008.
L. Pan, S. Alagapan, E. Franca, G. J. Brewer, and B. C. Wheeler, Propagation of action potential activity in a predefined microtunnel neural network, Journal of neural engineering, vol. 8, p. 046031, 2011.
J. W. Park, H. J. Kim, M. W. Kang, and N. L. Jeon, Advances in microfluidics-based experimental methods for neuroscience research, Lab on a Chip, vol. 13, pp. 509-521, 2013.
Paxinos, G. and C. Watson, „The rat brain in stereotaxic coordinates,“ Academic press, 2007.
J. Pine, “Recording action potentials from cultured neurons with extracellular microcircuit electrodes, Journal of neuroscience methods, vol. 2, no. 1, pp. 19-31, 1980.
V. S. Polikov, P. A. Tresco, and W. M. Reichert, Response of brain tissue to chronically implanted neural electrodes, Journal of neuroscience methods, vol. 148, pp. 1-18, 2005.
A. M. Pooler, W. Noble, and D. P. Hanger, A role for tau at the synapse in Alzheimer's disease pathogenesis, Neuropharmacology, vol. 76, pp. 1-8, 2014.
S. M. Potter and T. B. DeMarse, A new approach to neural cell culture for long-term studies, Journal of neuroscience methods, vol. 110, pp. 17-24, 2001.
A. Probst, J. Gotz, K. H. Wiederhold, M. Tolnay, C. Mistl, A. L. Jaton, M. Hong, T. Ishihara, V.-Y. Lee, and J. Q. Trojanowski, Axonopathy and amyotrophy in mice transgenic for human four-repeat tau protein, Acta neuropathologica, vol. 99, pp. 469-481, 2000.
Y. Qiu, R. Liao, and X. Zhang, Real-time monitoring primary cardiomyocyte adhesion based on electrochemical impedance spectroscopy and electrical cell-substrate impedance sensing, Analytical chemistry, vol. 80, pp. 990-996, 2008.
Y. Qiu, R. Liao, and X. Zhang, Impedance-Based Monitoring of Ongoing Cardiomyocyte Death Induced by Tumor Necrosis Factor-a, Biophysical journal, vol. 96, pp. 1985-1991, 2009.
F. Rehfeldt, A. J. Engler, A. Eckhardt, F. Ahmed, and D. E. Discher, Cell responses to the mechanochemical microenvironment- implications for regenerative medicine and drug delivery, Advanced drug delivery reviews, vol. 59, pp. 1329-1339, 2007.
E. D. Roberson, K. Scearce-Levie, J. J. Palop, F. Yan, I. H. Cheng, T. Wu, H. Gerstein, G.-Q. Yu, and L. Mucke, Reducing endogenous tau ameliorates amyloid ?-induced deficits in an Alzheimer's disease mouse model, Science, vol. 316, pp. 750-754, 2007.
A. B. Rocher, J. L. Crimins, J. M. Amatrudo, M. S. Kinson, M. A. Todd-Brown, J. Lewis, and J. I. Luebke, Structural and functional changes in tau mutant mice neurons are not linked to the presence of NFTs, Experimental neurology, vol. 223, pp. 385-393, 2010.
W. Rutten, J. M. Mouveroux, J. Buitenweg, C. Heida, T. Ruardij, E. Marani, and E. Lakke, Neuroelectronic interfacing with cultured multielectrode arrays toward a cultured probe, Proceedings of the IEEE, vol. 89, pp. 1013-1029, 2001.
S. Saman, W. Kim, M. Raya, Y. Visnick, S. Miro, S. Saman, B. Jackson, A. C. McKee, V. E. Alvarez, and N. C. Y. Lee, Exosome-associated tau is secreted in tauopathy models and is selectively phosphorylated in cerebrospinal fluid in early Alzheimer disease, Journal of Biological Chemistry, vol. 287, pp. 3842-3849, 2012.
G. Shahaf and S. Marom, Learning in networks of cortical neurons, The Journal of Neuroscience, vol. 21, pp. 8782-8788, 2001.
D. H. Small, S. San Mok, and J. C. Bornstein, Alzheimer's disease and Aβ toxicity: from top to bottom, Nature Reviews Neuroscience, vol. 2, pp. 595-598, 2001.
D. H. Small, Network dysfunction in Alzheimer's disease: does synaptic scaling drive disease progression?, Trends in molecular medicine, vol. 14, pp. 103-108, 2008.
C. Spegel, A. Heiskanen, L. H. D. Skjolding and Jenny Emnéus., “Chip based electroanalytical systems for cell analysis, Electroanalysis, vol. 20, no. 6, pp. 680-702, 2008.
K. Stamer, R. Vogel, E. Thies, E. Mandelkow, and E. M. Mandelkow, Tau blocks traffic of organelles, neurofilaments, and APP vesicles in neurons and enhances oxidative stress, The Journal of cell biology, vol. 156, pp. 1051-1063, 2002.
A. Stett, U. Egert, E. Guenther, F. Hofmann, T. Meyer, Wi. Nisch, and H. Haemmerle, “Biological application of microelectrode arrays in drug discovery and basic research, Analytical and bioanalytical chemistry, vol. 377, no. 3, pp. 486-495, 2003.
G. B. Stokin, C. Lillo, T. L. Falzone, R. G. Brusch, E. Rockenstein, S. L. Mount, R. Raman, P. Davies, E. Masliah, and D. S. Williams, Axonopathy and transport deficits early in the pathogenesis of Alzheimer's disease, Science, vol. 307, pp. 1282-1288, 2005.
E. Storey, G. J. Kinsella, and M. J. Slavin, The neuropsychological diagnosis of Alzheimer's disease, Journal of Alzheimer's Disease, vol. 3, pp. 261-285, 2001.
Su F.C., C. C. Wu, S. Chien, Review: roles of microenvironment and mechanical forces in cell and tissue remodeling, Journal of Medical and Biological Engineering, vol. 31,pp. 233-244, 2011.
J. Subbaroyan, D. C. Martin, and D. R. Kipke, A finite-element model of the mechanical effects of implantable microelectrodes in the cerebral cortex, Journal of neural engineering, vol. 2, p. 103, 2005.
A. Sydow, A. Van der Jeugd, F. Zheng, T. Ahmed, D. Balschun, O. Petrova, D. Drexler, L. Zhou, G. Rune, and E. Mandelkow, Tau-induced defects in synaptic plasticity, learning, and memory are reversible in transgenic mice after switching off the toxic Tau mutant, The Journal of Neuroscience, vol. 31, pp. 2511-2525, 2011.
D. H. Szarowski, M. D. Andersen, S. Retterer, A. J. Spence, M. Isaacson, H. G. Craighead, J. N. Turner, and W. Shain, Brain responses to micro-machined silicon devices, Brain research, vol. 983, pp. 23-35, 2003.
S. Takeuchi, D. Ziegler, Y. Yoshida, K. Mabuchi, and T. Suzuki, Parylene flexible neural probes integrated with microfluidic channels, Lab on a Chip, vol. 5, pp. 519-523, 2005.
A. M. Taylor, M. Blurton-Jones, S. W. Rhee, D. H. Cribbs, C. W. Cotman, and N. L. Jeon, A microfluidic culture platform for CNS axonal injury, regeneration and transport, Nature methods, vol. 2, pp. 599-605, 2005.
A. M. Taylor, D. C. Dieterich, H. T. Ito, S. A. Kim, and E. M. Schuman, Microfluidic local perfusion chambers for the visualization and manipulation of synapses, Neuron, vol. 66, pp. 57-68, 2010.
E. Thies and E.-M. Mandelkow, Missorting of tau in neurons causes degeneration of synapses that can be rescued by the kinase MARK2/Par-1, The Journal of neuroscience, vol. 27, pp. 2896-2907, 2007.
L. W. Tien, F. Wu, M. D. Tang-Schomer, E. Yoon, F. G. Omenetto, and D. L. Kaplan, Silk as a Multifunctional Biomaterial Substrate for Reduced Glial Scarring around Brain-Penetrating Electrodes, Advanced Functional Materials, vol. 23, pp. 3185-3193.
M. Tijero, G. Gabriel, J. Caro, A. Altuna, R. Hernandez, R. Villa, J. Berganzo, F. J. Blanco, R. Salido, and L. J. Fernandez, SU-8 microprobe with microelectrodes for monitoring electrical impedance in living tissues, Biosensors and Bioelectronics, vol. 24, pp. 2410-2416, 2009.
C. Tiruppathi, A. B. Malik, P. J. Del Vecchio, C. R. Keese, and I. Giaever, Electrical method for detection of endothelial cell shape change in real time: assessment of endothelial barrier function, Proceedings of the National Academy of Sciences, vol. 89, pp. 7919-7923, 1992.
J. N. Turner, W. Shain, D. H. Szarowski, M. Andersen, S. Martins, M. Isaacson, and H. Craighead, Cerebral astrocyte response to micromachined silicon implants, Experimental neurology, vol. 156, pp. 33-49, 1999.
I. S. Uroukov and L. Bull, On the effect of long-term electrical stimulation on three-dimensional cell cultures: Hen embryo brain spheroids, Medical devices (Auckland, NZ), vol. 1, p. 1, 2008.
T. Valero, G. Moschopoulou, S. Kintzios, P. Hauptmann, M. Naumann, and T. Jacobs, Studies on neuronal differentiation and signalling processes with a novel impedimetric biosensor, Biosensors and Bioelectronics, vol. 26, pp. 1407-1413, 2010.
J. van Pelt, P. S. Wolters, M. A. Corner et al., “Long-term characterization of firing dynamics of spontaneous bursts in cultured neural networks, Biomedical Engineering, IEEE Transactions on, vol. 51, no. 11, pp. 2051-2062, 2004.
V. N. Vernekar, D. K. Cullen, N. Fogleman, Y. Choi, A. J. Garcia, M. G. Allen, G. J. Brewer, and M. C. LaPlaca, SU - 2000 rendered cytocompatible for neuronal bioMEMS applications, Journal of Biomedical Materials Research Part A, vol. 89, pp. 138-151, 2009.
R. J. Vetter, J. C. Williams, J. F. Hetke, E. A. Nunamaker, and D. R. Kipke, Chronic neural recording using silicon-substrate microelectrode arrays implanted in cerebral cortex, Biomedical Engineering, IEEE Transactions on, vol. 51, pp. 896-904, 2004.
G. Voskerician, M. S. Shive, R. S. Shawgo, H. v. Recum, J. M. Anderson, M. J. Cima, and R. Langer, Biocompatibility and biofouling of MEMS drug delivery devices, Biomaterials, vol. 24, pp. 1959-1967, 2003.
J. Vukasinovic, D. K. Cullen, M. C. LaPlaca, and A. Glezer, A microperfused incubator for tissue mimetic 3D cultures, Biomedical microdevices, vol. 11, pp. 1155-1165, 2009.
P.-H. Wang, I. L. Tseng, and S.-h. Hsu, Review: bioengineering approaches for guided peripheral nerve regeneration, J. Med. Biol. Eng, vol. 31, pp. 151-160, 2011.
Y. Wang, G. Zhang, H. Zhou, A. Barakat, and H. Querfurth, Opposite effects of low and high doses of Aβ42 on electrical network and neuronal excitability in the rat prefrontal cortex, PloS one, vol. 4, p. e8366, 2009.
D. A. Wagenaar, J. Pine, and S. M. Potter, An extremely rich repertoire of bursting patterns during the development of cortical cultures, BMC neuroscience, vol. 7, p. 11, 2006.
M. P. Ward, P. Rajdev, C. Ellison, and P. P. Irazoqui, Toward a comparison of microelectrodes for acute and chronic recordings, Brain research, vol. 1282, pp. 183-200, 2009.
T. Ware, D. Simon, D. E. Arreaga-Salas, J. Reeder, R. Rennaker, E. W. Keefer, and W. Voit, Fabrication of responsive, softening neural interfaces, Advanced Functional Materials, vol. 22, pp. 3470-3479, 2012.
J. Wegener, C. R. Keese, and I. Giaever, Electric cell-ubstrate impedance sensing (ECIS) as a noninvasive means to monitor the kinetics of cell spreading to artificial surfaces, Experimental cell research, vol. 259, pp. 158-166, 2000.
S. M. Willerth, K. J. Arendas, D. I. Gottlieb, and S. E. Sakiyama-Elbert, Optimization of fibrin scaffolds for differentiation of murine embryonic stem cells into neural lineage cells, Biomaterials, vol. 27, pp. 5990-6003, 2006.
D. F. Williams, On the mechanisms of biocompatibility, Biomaterials, vol. 29, pp. 2941-2953, 2008.
J. C. Williams, R. L. Rennaker, and D. R. Kipke, Long-term neural recording characteristics of wire microelectrode arrays implanted in cerebral cortex, Brain Research Protocols, vol. 4, pp. 303-313, 1999.
G. Xiang, L. Pan, L. Huang, Z. Yu, X. Song, J. Cheng, W. Xing, and Y. Zhou, Microelectrode array-based system for neuropharmacological applications with cortical neurons cultured in vitro, Biosensors and Bioelectronics, vol. 22, pp. 2478-2484, 2007.
C. Xiao, B. Lachance, G. Sunahara, and J. H. T. Luong, Assessment of cytotoxicity using electric cell-substrate impedance sensing: concentration and time response function approach, Analytical chemistry, vol. 74, pp. 5748-5753, 2002.
C. Xiao and J. H. T. Luong, On-line monitoring of cell growth and cytotoxicity using electric cell-substrate impedance sensing (ECIS), Biotechnology progress, vol. 19, pp. 1000-1005, 2003.
C. Xiao and J. H. T. Luong, Assessment of cytotoxicity by emerging impedance spectroscopy, Toxicology and applied pharmacology, vol. 206, pp. 102-112, 2005.
T. Xu, P. Molnar, C. Gregory, M. Das, T. Boland, and J. J. Hickman, Electrophysiological characterization of embryonic hippocampal neurons cultured in a 3D collagen hydrogel, Biomaterials, vol. 30, pp. 4377-4383, 2009.
M. Yang, C. C. Lim, R. Liao, and X. Zhang, A novel microfluidic impedance assay for monitoring endothelin-induced cardiomyocyte hypertrophy, Biosensors and Bioelectronics, vol. 22, pp. 1688-1693, 2007.
H. Yin, F. L. Wang, A. L. Wang, J. Cheng, and Y. Zhou, Bioelectrical Impedance Assay to Monitor Changes in Aspirin-treated Human Colon Cancer HT9 Cell Shape during Apoptosis, Analytical letters, vol. 40, pp. 85-94, 2007.
E. Yoshida, T. G. Atkinson, and B. Chakravarthy, Neuroprotective gene expression profiles in ischemic cortical cultures preconditioned with IGF-1 or bFGF, Molecular brain research, vol. 131, pp. 33-50, 2004.
S. Zhang, F. Gelain, and X. Zhao, Designer self-assembling peptide nanofiber scaffolds for 3D tissue cell cultures, Seminars in cancer biology, pp. 413-420, 2005.

Z. Zhang, P. Zhao, and G. Xiao, The fabrication of polymer microfluidic devices using a solid-to-solid interfacial polyaddition, Polymer, vol. 50, pp. 5358-5361, 2009.
Z. Zhang and J. W. Simpkins, An okadaic acid-induced model of tauopathy and cognitive deficiency, Brain research, vol. 1359, pp. 233-246, 2010.