Electrospun Poly(benzyl glutamate)-r-Poly(glutamic acid) Scaffold for PC-12 Neurite outgrowth
Date Issued
2016
Date
2016
Author(s)
Wang, Zhen-Hua
Abstract
As portable electronic devices popularize, people seems to be addicted to them for getting new information or watching videos. However, using eye for long time that causes optic nerve damage. Clinical research reports that macular degeneration and glaucoma have been found on people in their thirties more often than before which may be using electronic devices without proper distance and adequate eye relaxation. As long as optic nerve is damaged, it will not regenerate by itself. We would like to culture optic nerves in vitro on scaffold by tissue engineering, then transfer the scaffold to damaged part of nerves for regeneration. In tissue engineering, the scaffold must be porous, cell-recognized motif, biodegradable, biocompatible which is mimic native tissue. Electrospin is a promising technique to fabricate nanofibrous scaffold which is mimic the structure of extracellular matrix (ECM) and provides high surface area with interconnected pores. Poly(benzyl glutamate)-r-poly(glutamic acid) (PBGA) have glutamic acid, which is a excitatory neurotransmitter that plays the principal role in neural activation. We expect it can benefit neuron differentiation by glutamic acid motif during cell culture. In this study, we have synthesized monomer: γ-benzyl glutamate-N-carboxy anhydride from L-glutamic acid γ-benzyl ester and triphosgene. Then we used sodium methoxide as an initiator to polymerize the monomer into poly(benzyl-glutamate) in anhydrous benzene. Finally, we controlled the ratio of glutamic acid on the main chain by hydrolysis time. After getting product, we use NMR and GPC to analyze the ratio of glutamic acid in copolymer and molecular weight of PBGA. The PBG and PBGA were further fabricated into fibrous scaffolds by electrospinning using tetrahydrofuran (THF) and dimethylacetamide (DMAc) cosolvent. We investigated the effect of glutamic acid ratio on PC-12 cells proliferation and neuron differentiation. We found that the biocompatibility and neuron differentiation can be improved by glutamic acid. In our study, The copolymer contained 30% molar ratio of glutamic acid (PBGA30) exhibited best biocompatibility and the longest nerve outgrowth. However, neuron transmit direction plays important role on nerve tissue functionality. We fabricated scaffold with aligned fibers by employing a rotating drum collector to guide the growth of nerve. We found that neurite is growth along the fiber. In summary, the aligned electrospun PBGA scaffold has potential for optic nerve regeneration.
Subjects
poly(benzyl-glutamate)-r-poly(glutamic acid)
electrospinning
PC-12 culture
optic nerve regeneration
Type
thesis
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