Multi-walled carbon nanotube

(CNT) arrays with chemical modifications and 3D nanotopography greatly enhanced the adhesion and organization of the functional neuronal network [10, 11]. Positively charged nanofibers dictated neuron adhesion and network formation [12]. CNT clusters promoted complex and interconnected neuronal network formation via the self-assembly process of neurons [13, 14]. Topography affects the growth direction of processes and the adhesion of astrocytes. Nanotopography might affect the constructs and functions of astrocytes, leading to the regulation of hyperexcitability and epileptic activity in neurons. Structures with topographic patterns can control cell behavior, and the interactions between SCH727965 mouse cells and substrates may play an important role in substrate biocompatibility [15]. However, the effects of glial-substrate interactions on the astrocytic syncytium are not clear. In this report, we used ordered nanotopography to study the molecular mechanisms underlying topographic control of the astrocytic syncytium of the C6 glioma. Nanotopography is capable of modulating transport of gap Danusertib junction protein and influencing the cell-cell interactions of astrocytes. Methods Cell culture The C6 glioma-astrocytoma rat cell line, C6.51.passage, was purchased from the Bioresource Collection and Research Center

(BCRC; Hsinchu, Taiwan). C6 cells were cultured in Hamćs F10 medium with sodium bicarbonate (NaHCO3), horse serum (HS), fetal bovine serum (FBS), GlutaMAX I (Thermo Fisher Scientific Inc., Waltham, MA, USA), trypsin, and BSA (bovine serum albumin), which were purchased from GIBCO (Thermo find more Fisher

Scientific Inc.). The cells were Chloroambucil incubated at 37°C in 5% CO2. Chemicals A CellTiter 96® AQueous One Solution Cell Proliferation Assay (MTS assay) was purchased from Promega (Madison, WI, USA). Phosphate-buffered saline (PBS) was purchased from Bio-tech (Taipei, Taiwan). Anti-vinculin antibody (vinculin) and anti-connexin43 antibody (connexin43) were purchased from Abcam (Cambridge, England, UK) and Invitrogen (Renfrew, UK), respectively. Anti-glial fibrillary acidic protein antibody (GFAP), luminol reagent, and oxidizing reagent were purchased from Millipore (Billerica, MA, USA). Sulfuric acid (H2SO4), oxalic acid (H2C2O4), and phosphoric acid (H3PO4) were purchased from Sigma Chemicals (Perth, Western Australia). Other chemicals of analytical grade or higher were purchased from Sigma or Millipore. Fabrication of nanodot surfaces Nanodot arrays were fabricated as previously described [16]. A 200-nm-thick tantalum nitride (TaN) thin film was sputtered onto a 6-in silicon wafer (Summit-Tech, West Hartford, CT, USA), followed by a deposition of a 400-nm-thick aluminum (Admat-Midas, Norristown, PA, USA) layer on top of the TaN thin film. Anodization was performed using either 1.8 M H2SO4 at 5 V for 1.5 h (for the 10-nm nanodot array) or 0.