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On-line Access: 2022-01-12

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 ORCID:

Shengbo SANG

https://orcid.org/0000-0003-3011-7632

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Journal of Zhejiang University SCIENCE B 2022 Vol.23 No.1 P.58-73

http://doi.org/10.1631/jzus.B2100155


Biocompatible chitosan/polyethylene glycol/multi-walled carbon nanotube composite scaffolds for neural tissue engineering


Author(s):  Shengbo SANG, Rong CHENG, Yanyan CAO, Yayun YAN, Zhizhong SHEN, Yajing ZHAO, Yanqing HAN

Affiliation(s):  Shanxi Key Laboratory of Micro Nano Sensors & Artificial Intelligence Perception, College of Information and Computer, Taiyuan University of Technology, Taiyuan 030024, China; more

Corresponding email(s):   sunboa-sang@tyut.edu.cn

Key Words:  Multi-walled carbon nanotube (MWCNT), Cell-scaffold, PC12 cells, Biocompatibility


Shengbo SANG, Rong CHENG, Yanyan CAO, Yayun YAN, Zhizhong SHEN, Yajing ZHAO, Yanqing HAN. Biocompatible chitosan/polyethylene glycol/multi-walled carbon nanotube composite scaffolds for neural tissue engineering[J]. Journal of Zhejiang University Science B, 2022, 23(1): 58-73.

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author="Shengbo SANG, Rong CHENG, Yanyan CAO, Yayun YAN, Zhizhong SHEN, Yajing ZHAO, Yanqing HAN",
journal="Journal of Zhejiang University Science B",
volume="23",
number="1",
pages="58-73",
year="2022",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B2100155"
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%T Biocompatible chitosan/polyethylene glycol/multi-walled carbon nanotube composite scaffolds for neural tissue engineering
%A Shengbo SANG
%A Rong CHENG
%A Yanyan CAO
%A Yayun YAN
%A Zhizhong SHEN
%A Yajing ZHAO
%A Yanqing HAN
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%DOI 10.1631/jzus.B2100155

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T1 - Biocompatible chitosan/polyethylene glycol/multi-walled carbon nanotube composite scaffolds for neural tissue engineering
A1 - Shengbo SANG
A1 - Rong CHENG
A1 - Yanyan CAO
A1 - Yayun YAN
A1 - Zhizhong SHEN
A1 - Yajing ZHAO
A1 - Yanqing HAN
J0 - Journal of Zhejiang University Science B
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PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B2100155


Abstract: 
Carbon nanotube (CNT) composite materials are very attractive for use in neural tissue engineering and biosensor coatings. CNT scaffolds are excellent mimics of extracellular matrix due to their hydrophilicity, viscosity, and biocompatibility. CNTs can also impart conductivity to other insulating materials, improve mechanical stability, guide neuronal cell behavior, and trigger axon regeneration. The performance of chitosan (CS)/polyethylene glycol (PEG) composite scaffolds could be optimized by introducing multi-walled CNTs (MWCNTs). CS/PEG/CNT composite scaffolds with CNT content of 1%, 3%, and 5% (1%=0.01 g/mL) were prepared by freeze-drying. Their physical and chemical properties and biocompatibility were evaluated. Scanning electron microscopy (SEM) showed that the composite scaffolds had a highly connected porous structure. Transmission electron microscope (TEM) and Raman spectroscopy proved that the CNTs were well dispersed in the CS/PEG matrix and combined with the CS/PEG nanofiber bundles. MWCNTs enhanced the elastic modulus of the scaffold. The porosity of the scaffolds ranged from 83% to 96%. They reached a stable water swelling state within 24 h, and swelling decreased with increasing MWCNT concentration. The electrical conductivity and cell adhesion rate of the scaffolds increased with increasing MWCNT content. Immunofluorescence showed that rat pheochromocytoma (PC12) cells grown in the scaffolds had characteristics similar to nerve cells. We measured changes in the expression of nerve cell markers by quantitative real-time polymerase chain reaction (qRT-PCR), and found that PC12 cells cultured in the scaffolds expressed growth-associated protein 43 (GAP43), nerve growth factor receptor (NGFR), and class III β‍-tubulin (TUBB3) proteins. Preliminary research showed that the prepared CS/PEG/CNT scaffold has good biocompatibility and can be further applied to neural tissue engineering research.

应用于神经组织工程的生物相容性壳聚糖/聚乙二醇/多壁碳纳米管复合支架的研究

目的:对壳聚糖/聚乙二醇/多壁碳纳米管(CS/PEG/MWCNT)复合支架的理化性质及对神经细胞的生物相容性进行研究,探究碳纳米管(CNT)的最佳浓度,同时判断其是否可以进一步应用于神经组织工程研究。
创新点:通过引入MWCNT来优化CS/PEG复合支架的综合性能。
方法:通过冷冻干燥制备CNTs的质量体积分数(g/mL)分别为0%、1%、3%和5%的CS/PEG/CNT神经复合支架,对其进行一系列的物理化学性质的表征,包括扫描电镜(SEM)、透射电镜(TEM)、红外光谱、拉曼光谱、杨氏模量、孔隙率、水溶胀以及导电率等。进而将神经细胞接种到支架上进行体外培养,通过对支架上的细胞增殖及粘附、活/死细胞染色来观察所制备神经支架的细胞毒性,通过免疫荧光及实时定量聚合酶链式反应(qRT-PCR)检测了神经细胞标志物表达的变化,进一步探究不同支架对神经细胞分化的影响。
结论:CS/PEG/CNTs支架表现出紧密而整洁的中空网络结构,且CNTs很好地分散在CS/PEG基质中。CNTs的添加增加了支架孔壁的粗糙度、导电性和疏水性,提高了支架的力学性能,降低了溶胀率和生物降解率。PC12细胞体外生物学评价结果表明:CS/PEG/CNTs支架具有良好的生物相容性,无细胞毒性,增强了体外神经元细胞的增殖和分化能力。总体而言,CS/PEG/3%CNTs支架具有优异的综合性能,有望用作周围神经再生的神经元生长基质。

关键词:多壁碳纳米管(MWCNTs);神经细胞支架;PC12细胞;生物相容性

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

[1]Abidian MR, Daneshvar ED, Egeland BM, et al., 2012. Hybrid conducting polymer—hydrogel conduits for axonal growth and neural tissue engineering. Adv Healthc Mater, 1(6):762-767.

[2]Ahn HS, Hwang JY, Kim SM, et al., 2015. Carbon-nanotube-interfaced glass fiber scaffold for regeneration of transected sciatic nerve. Acta Biomater, 13:324-334.

[3]Bhaskar B, Owen R, Bahmaee H, et al., 2018. Composite porous scaffold of PEG/PLA support improved bone matrix deposition in vitro compared to PLA-only scaffolds. J Biomed Mater Res A, 106(5):1334-1340.

[4]Bosi S, Ballerini L, Prato M, 2014. Carbon nanotubes in tissue engineering. In: Marcaccio M, Paolucci F (Eds.), Making and Exploiting Fullerenes, Graphene, and Carbon Nanotubes. Springer, Berlin, p.181-204.

[5]Duan B, Gao HM, He M, et al., 2014. Hydrophobic modification on surface of chitin sponges for highly effective separation of oil. ACS Appl Mater Interfaces, 6(22):19933-19942.

[6]Eldesoqi K, Henrich D, El-Kady AM, et al., 2014. Safety evaluation of a bioglass—polylactic acid composite scaffold seeded with progenitor cells in a rat skull critical-size bone defect. PLoS ONE, 9(2):e87642.

[7]Fabbro A, Sucapane A, Toma FM, et al., 2013a. Adhesion to carbon nanotube conductive scaffolds forces action-potential appearance in immature rat spinal neurons. PLoS ONE, 8(8):e73621.

[8]Fabbro A, Prato M, Ballerini L, 2013b. Carbon nanotubes in neuroregeneration and repair. Adv Drug Deliv Rev, 65(15): 2034-2044.

[9]Gao JJ, Zhu J, Luo JJ, et al., 2016. Investigation of microporous composite scaffolds fabricated by embedding sacrificial polyethylene glycol microspheres in nanofibrous membrane. Compos Part A Appl Sci Manuf, 91:20-29.

[10]Ghasemi-Mobarakeh L, Prabhakaran MP, Morshed M, et al., 2011. Application of conductive polymers, scaffolds and electrical stimulation for nerve tissue engineering. J Tissue Eng Regen Med, 5(4):e17-e35.

[11]Gu XS, Ding F, Williams DF, 2014. Neural tissue engineering options for peripheral nerve regeneration. Biomaterials, 35(24):6143-6156.

[12]He M, Wang ZG, Cao Y, et al., 2014. Construction of chitin/PVA composite hydrogels with jellyfish gel-like structure and their biocompatibility. Biomacromolecules, 15(9):3358-3365.

[13]Huang Y, Zhong ZB, Duan B, et al., 2014. Novel fibers fabricated directly from chitin solution and their application as wound dressing. J Mater Chem B, 2(22):3427-3432.

[14]Ifuku S, Saimoto H, 2012. Chitin nanofibers: preparations, modifications, and applications. Nanoscale, 4(11):3308-3318.

[15]Jayakumar R, Prabaharan M, Kumar PTS, et al., 2011. Biomaterials based on chitin and chitosan in wound dressing applications. Biotechnol Adv, 29(3):322-337.

[16]Ji CD, Annabi N, Hosseinkhani M, et al., 2012. Fabrication of poly-DL-lactide/polyethylene glycol scaffolds using the gas foaming technique. Acta Biomater, 8(2):570-578.

[17]Kotturi H, Abuabed A, Zafar H, et al., 2017. Evaluation of polyethylene glycol diacrylate-polycaprolactone scaffolds for tissue engineering applications. J Funct Biomater, 8(3):39.

[18]Lau C, Cooney MJ, Atanassov P, 2008. Conductive macroporous composite chitosan-carbon nanotube scaffolds. Langmuir, 24(13):7004-7010.

[19]Liu XF, Miller AL II, Park S, et al., 2016. Covalent crosslinking of graphene oxide and carbon nanotube into hydrogels enhances nerve cell responses. J Mater Chem B, 4(43):6930-6941.

[20]Ma SQ, Chen Z, Qiao F, et al., 2014. Guided bone regeneration with tripolyphosphate cross-linked asymmetric chitosan membrane. J Dent, 42(12):1603-1612.

[21]Mattioli-Belmonte M, Vozzi G, Whulanza Y, et al., 2012. Tuning polycaprolactone—carbon nanotube composites for bone tissue engineering scaffolds. Mater Sci Eng C, 32(2):152-159.

[22]Mauro N, Manfredi A, Ranucci E, et al., 2013. Degradable poly (amidoamine) hydrogels as scaffolds for in vitro culturing of peripheral nervous system cells. Macromol Biosci, 13(3):332-347.

[23]Mehdikhani M, Ghaziof S, 2018. Electrically conductive poly-‍ε‍-caprolactone/polyethylene glycol/multi-wall carbon nanotube nanocomposite scaffolds coated with fibrin glue for myocardial tissue engineering. Appl Phys A, 124:77.

[24]Nagarajan S, Belaid H, Pochat-Bohatier C, et al., 2017. Design of boron nitride/gelatin electrospun nanofibers for bone tissue engineering. ACS Appl Mater Interfaces, 9(39):33695-33706.

[25]Naumenko E, Fakhrullin R, 2019. Halloysite nanoclay/biopolymers composite materials in tissue engineering. Biotechnol J, 14(12):1900055.

[26]Nisbet DR, Crompton KE, Horne MK, et al., 2008. Neural tissue engineering of the CNS using hydrogels. J Biomed Mater Res Part B Appl Biomater, 87B(1):251-263.

[27]Park SY, Choi DS, Jin HJ, et al., 2011. Polarization-controlled differentiation of human neural stem cells using synergistic cues from the patterns of carbon nanotube monolayer coating. ACS Nano, 5(6):4704-4711.

[28]Posypanova GA, Gayduchenko IA, Moskaleva EY, et al., 2016. Neuronal differentiation of PC12 cells and mouse neural stem cells on carbon nanotube films. Cell Tissue Biol, 10(3):194-201.

[29]Rad SM, Khorasani MT, Joupari MD, 2016. Preparation of HMWCNT/PLLA nanocomposite scaffolds for application in nerve tissue engineering and evaluation of their physical, mechanical and cellular activity properties. Polym Adv Technol, 27(3):325-338.

[30]Runge MB, Dadsetan M, Baltrusaitis J, et al., 2010. Development of electrically conductive oligo(polyethylene glycol) fumarate-polypyrrole hydrogels for nerve regeneration. Biomacromolecules, 11(11):2845-2853.

[31]ŞenÖ, Culha M, 2016. Boron nitride nanotubes included thermally cross-linked gelatin—glucose scaffolds show improved properties. Colloids Surf B Biointerfaces, 138:41-49.

[32]Shitole AA, Giram PS, Raut PW, et al., 2019. Clopidogrel eluting electrospun polyurethane/polyethylene glycol thromboresistant, hemocompatible nanofibrous scaffolds. J Biomater Appl, 33(10):1327-1347.

[33]Shokrgozar MA, Mottaghitalab F, Mottaghitalab V, et al., 2011. Fabrication of porous chitosan/poly(vinyl alcohol) reinforced single-walled carbon nanotube nanocomposites for neural tissue engineering. J Biomed Nanotechnol, 7(2):276-284.

[34]Srinivasan A, Teo N, Poon KJ, et al., 2021. Comparative craniofacial bone regeneration capacities of mesenchymal stem cells derived from human neural crest stem cells and bone marrow. ACS Biomater Sci Eng, 7(1):207-221.

[35]Stock K, Nolden L, Edenhofer F, et al., 2010. Transcription factor-based modulation of neural stem cell differentiation using direct protein transduction. Cell Mol Life Sci, 67(14):2439-2449.

[36]Suner SS, Demirci S, Yetiskin B, et al., 2019. Cryogel composites based on hyaluronic acid and halloysite nanotubes as scaffold for tissue engineering. Int J Biol Macromol, 130:627-635.

[37]Teo AJT, Mishra A, Park I, et al., 2016. Polymeric biomaterials for medical implants and devices. ACS Biomater Sci Eng, 2(4):454-472.

[38]Türk S, Altınsoy I, Efe GÇ, et al., 2018. 3D porous collagen/functionalized multiwalled carbon nanotube/chitosan/hydroxyapatite composite scaffolds for bone tissue engineering. Mater Sci Eng C, 92:757-768.

[39]Uto K, Mano SS, Aoyagi T, et al., 2016. Substrate fluidity regulates cell adhesion and morphology on poly(ε‍-‍caprolactone)-based materials. ACS Biomater Sci Eng, 2(3):446-453.

[40]van den Broeck L, Piluso S, Soultan AH, et al., 2019. Cytocompatible carbon nanotube reinforced polyethylene glycol composite hydrogels for tissue engineering. Mater Sci Eng C, 98:1133-1144.

[41]Venkatesan J, Ryu BM, Sudha PN, et al., 2012. Preparation and characterization of chitosan-carbon nanotube scaffolds for bone tissue engineering. Int J Biol Macromol, 50(2):393-402.

[42]Wang JY, Sun PP, Bao YM, et al., 2011. Cytotoxicity of single-walled carbon nanotubes on PC12 cells. Toxicol in Vitro, 25(1):242-250.

[43]Wu J, Meredith JC, 2014. Assembly of chitin nanofibers into porous biomimetic structures via freeze drying. ACS Macro Lett, 3(2):185-190.

[44]Wu SQ, Duan B, Zeng XP, et al., 2017. Construction of blood compatible lysine-immobilized chitin/carbon nanotube microspheres and potential applications for blood purified therapy. J Mater Chem B, 5(16):2952-2963.

[45]Xu DF, Fan L, Gao LF, et al., 2016. Micro-nanostructured polyaniline assembled in cellulose matrix via interfacial polymerization for applications in nerve regeneration. ACS Appl Mate Interfaces, 8(27):17090-17097.

[46]Zhou ZF, Liu XF, Wu W, et al., 2018. Effective nerve cell modulation by electrical stimulation of carbon nanotube embedded conductive polymeric scaffolds. Biomater Sci, 6(9):2375-2385.

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