title: Guidance of neural regeneration on the biomimetic nanostructured matrix.
abstract: Biomimetic materials are used for creating microsystems to control cell growth spatially and elicit specific cellular responses by combining complex biomolecules with nanostructured surfaces. Intercellular cell-to-cell and cell-to-extracellular matrix (ECM) interactions in biomimetic materials have demonstrated potential in the development of drug discovery platforms and regeneration medicine. In this study, we developed a biomimetic nanostructured matrix by using various ECM molecular layers to create a biomimetic and biocompatible environment for realizing neuronal guidance in neural regeneration medicine. Silicon-based substrates possessing nanostructures were modified using different ECM proteins and peptides to develop a biomimetic and biocompatible environment for studying neural behaviors in adhesion, proliferation, and differentiation. The substrates were flat glass, flat silicon wafers (FWs), and nanorod-structured wafers prepared using wet etching. The three substrates were then functionalized using laminin-1 peptide, PA22-2-contained active isoleucine-lysine-valine-alanine-valine (IKVAV) peptide, and poly-d-lysine (PDL), separately. When PC12 cells were cultured and differentiated on the modified substrates, the cells were able to elongate the neurites on the glass and FW, which was coated with three types of peptide. More differentiated neurons were observed on the nanorod-structured wafers coated with laminin than on those coated with IKVAV or PDL. For achieving directional guidance of neurite outgrowth, substrates exhibiting a grating pattern of nanorods were partially collapsed by the pulling force of water, leaving few nanorods, which support the net form of laminin on the surface. Furthermore, we fabricated the topological nanostructure-patterned wafer coated with laminin and successfully manipulated the extension and direction of neurites by using more than 80 μm of a single soma. This approach demonstrates potential as a facile and efficient method for guiding the direction of single axons and for enhancing neurite outgrowth in studies on nerve regenerative medicine.
authors: Instrument Technology Research Center, National Applied Research Laboratories, Hsinchu, Taiwan.
authors: Yen-Pei
authors: YP
authors: Lu
authors: Department of Biological Science and Technology, I-Shou University, Kaohsiung, Taiwan.
authors: Chih-Hui
authors: CH
authors: Yang
authors: Instrument Technology Research Center, National Applied Research Laboratories, Hsinchu, Taiwan; Institute of NanoEngineering and MicroSystems, National Tsing Hua University, Hsinchu, Taiwan.
authors: J Andrew
authors: JA
authors: Yeh
authors: Instrument Technology Research Center, National Applied Research Laboratories, Hsinchu, Taiwan.
authors: Fu Han
authors: FH
authors: Ho
authors: Instrument Technology Research Center, National Applied Research Laboratories, Hsinchu, Taiwan.
authors: Yu-Cheng
authors: YC
authors: Ou
authors: Department of Urology and Uro-oncology, China Medical University Beigang Hospital, Yunlin, Taiwan; Department of BioMedical Engineering, National Cheng Kung University, Tainan, Taiwan.
authors: Chieh Hsiao
authors: CH
authors: Chen
authors: Instrument Technology Research Center, National Applied Research Laboratories, Hsinchu, Taiwan. Electronic address: minyulin@itrc.narl.org.tw.
authors: Ming-Yu
authors: MY
authors: Lin
authors: The School of Chinese Medicine for Post-Baccalaureate, I-Shou University, Kaohsiung, Taiwan. Electronic address: huangks@isu.edu.tw.
authors: Keng-Shiang
authors: KS
authors: Huang
mesh_terms: D000818:Animals; D040761:Biomimetic Materials / Q000737:chemistry*; D005109:Extracellular Matrix; D007797:Laminin / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D043942:Nanotubes / Q000737:chemistry* / Q000648:ultrastructure; D009416:Nerve Regeneration; D016716:PC12 Cells; D010446:Peptide Fragments / Q000737:chemistry; D011107:Polylysine / Q000737:chemistry; D051381:Rats; D012825:Silicon / Q000737:chemistry*
chemical_list: D007797:Laminin; D010446:Peptide Fragments; C121346:laminin 1; C083321:isoleucyl-lysyl-valyl-alanyl-valine; D011107:Polylysine; D012825:Silicon
keywords: Biomimetic; Extracellular matrix; Nanostructure; Neural regeneration