Injectable Hydrogel Guides Neurons Growth with Specific Directionality

Visual disabilities affect more than 250 million people, with 43 million suffering from irreversible blindness. The eyes are an extension of the central nervous system which cannot regenerate. Neural tissue engineering is a potential method to cure the disease. Injectability is a desirable property...

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Main Authors: Yun-Hsiu Tseng, Tien-Li Ma, Dun-Heng Tan, An-Jey A. Su, Kia M. Washington, Chun-Chieh Wang, Yu-Ching Huang, Ming-Chung Wu, Wei-Fang Su
Format: Article
Language:English
Published: MDPI AG 2023-04-01
Series:International Journal of Molecular Sciences
Subjects:
Online Access:https://www.mdpi.com/1422-0067/24/9/7952
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author Yun-Hsiu Tseng
Tien-Li Ma
Dun-Heng Tan
An-Jey A. Su
Kia M. Washington
Chun-Chieh Wang
Yu-Ching Huang
Ming-Chung Wu
Wei-Fang Su
author_facet Yun-Hsiu Tseng
Tien-Li Ma
Dun-Heng Tan
An-Jey A. Su
Kia M. Washington
Chun-Chieh Wang
Yu-Ching Huang
Ming-Chung Wu
Wei-Fang Su
author_sort Yun-Hsiu Tseng
collection DOAJ
description Visual disabilities affect more than 250 million people, with 43 million suffering from irreversible blindness. The eyes are an extension of the central nervous system which cannot regenerate. Neural tissue engineering is a potential method to cure the disease. Injectability is a desirable property for tissue engineering scaffolds which can eliminate some surgical procedures and reduce possible complications and health risks. We report the development of the anisotropic structured hydrogel scaffold created by a co-injection of cellulose nanofiber (CNF) solution and co-polypeptide solution. The positively charged poly (L-lysine)-r-poly(L-glutamic acid) with 20 mol% of glutamic acid (PLLGA) is crosslinked with negatively charged CNF while promoting cellular activity from the acid nerve stimulate. We found that CNF easily aligns under shear forces from injection and is able to form hydrogel with an ordered structure. Hydrogel is mechanically strong and able to support, guide, and stimulate neurite growth. The anisotropy of our hydrogel was quantitatively determined in situ by 2D optical microscopy and 3D X-ray tomography. The effects of PLLGA:CNF blend ratios on cell viability, neurite growth, and neuronal signaling are systematically investigated in this study. We determined the optimal blend composition for stimulating directional neurite growth yielded a 16% increase in length compared with control, reaching anisotropy of 30.30% at 10°/57.58% at 30°. Using measurements of calcium signaling in vitro, we found a 2.45-fold increase vs. control. Based on our results, we conclude this novel material and unique injection method has a high potential for application in neural tissue engineering.
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spelling doaj.art-a39846de123f4e9ab78fa1fb0546893d2023-11-17T23:02:51ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-04-01249795210.3390/ijms24097952Injectable Hydrogel Guides Neurons Growth with Specific DirectionalityYun-Hsiu Tseng0Tien-Li Ma1Dun-Heng Tan2An-Jey A. Su3Kia M. Washington4Chun-Chieh Wang5Yu-Ching Huang6Ming-Chung Wu7Wei-Fang Su8Department of Materials Science and Engineering, National Taiwan University, Taipei 10617, TaiwanDepartment of Materials Science and Engineering, National Taiwan University, Taipei 10617, TaiwanDepartment of Materials Science and Engineering, National Taiwan University, Taipei 10617, TaiwanDepartment of Surgery, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USADepartment of Surgery, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USANational Synchrotron Radiation Research Center, Hsinchu 30076, TaiwanDepartment of Materials Engineering, Ming Chi University of Technology, New Taipei 24301, TaiwanDepartment of Chemical and Materials Engineering, Chang Gung University, Taoyuan 33302, TaiwanDepartment of Materials Science and Engineering, National Taiwan University, Taipei 10617, TaiwanVisual disabilities affect more than 250 million people, with 43 million suffering from irreversible blindness. The eyes are an extension of the central nervous system which cannot regenerate. Neural tissue engineering is a potential method to cure the disease. Injectability is a desirable property for tissue engineering scaffolds which can eliminate some surgical procedures and reduce possible complications and health risks. We report the development of the anisotropic structured hydrogel scaffold created by a co-injection of cellulose nanofiber (CNF) solution and co-polypeptide solution. The positively charged poly (L-lysine)-r-poly(L-glutamic acid) with 20 mol% of glutamic acid (PLLGA) is crosslinked with negatively charged CNF while promoting cellular activity from the acid nerve stimulate. We found that CNF easily aligns under shear forces from injection and is able to form hydrogel with an ordered structure. Hydrogel is mechanically strong and able to support, guide, and stimulate neurite growth. The anisotropy of our hydrogel was quantitatively determined in situ by 2D optical microscopy and 3D X-ray tomography. The effects of PLLGA:CNF blend ratios on cell viability, neurite growth, and neuronal signaling are systematically investigated in this study. We determined the optimal blend composition for stimulating directional neurite growth yielded a 16% increase in length compared with control, reaching anisotropy of 30.30% at 10°/57.58% at 30°. Using measurements of calcium signaling in vitro, we found a 2.45-fold increase vs. control. Based on our results, we conclude this novel material and unique injection method has a high potential for application in neural tissue engineering.https://www.mdpi.com/1422-0067/24/9/7952hydrogelpolypeptidecellulose nanofiberinjectablealigned structureneuron
spellingShingle Yun-Hsiu Tseng
Tien-Li Ma
Dun-Heng Tan
An-Jey A. Su
Kia M. Washington
Chun-Chieh Wang
Yu-Ching Huang
Ming-Chung Wu
Wei-Fang Su
Injectable Hydrogel Guides Neurons Growth with Specific Directionality
International Journal of Molecular Sciences
hydrogel
polypeptide
cellulose nanofiber
injectable
aligned structure
neuron
title Injectable Hydrogel Guides Neurons Growth with Specific Directionality
title_full Injectable Hydrogel Guides Neurons Growth with Specific Directionality
title_fullStr Injectable Hydrogel Guides Neurons Growth with Specific Directionality
title_full_unstemmed Injectable Hydrogel Guides Neurons Growth with Specific Directionality
title_short Injectable Hydrogel Guides Neurons Growth with Specific Directionality
title_sort injectable hydrogel guides neurons growth with specific directionality
topic hydrogel
polypeptide
cellulose nanofiber
injectable
aligned structure
neuron
url https://www.mdpi.com/1422-0067/24/9/7952
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AT dunhengtan injectablehydrogelguidesneuronsgrowthwithspecificdirectionality
AT anjeyasu injectablehydrogelguidesneuronsgrowthwithspecificdirectionality
AT kiamwashington injectablehydrogelguidesneuronsgrowthwithspecificdirectionality
AT chunchiehwang injectablehydrogelguidesneuronsgrowthwithspecificdirectionality
AT yuchinghuang injectablehydrogelguidesneuronsgrowthwithspecificdirectionality
AT mingchungwu injectablehydrogelguidesneuronsgrowthwithspecificdirectionality
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