电活性和生物活性多巴-胰岛素样生长因子-1@聚(乙交酯-丙交酯)/聚(3-己基噻吩)静电纺丝纤维的制备及神经组织工程应用

doi:10.11944/j.issn.1000-0518.2019.09.190027

摘要/Abstract

摘要：

理想型神经修复材料应具备与正常神经相似的导电性、仿生细胞外基质结构以及释放特定的生长因子等性能。本研究将不同质量分数(0、3%、5%、10%)的聚(3-己基噻吩)(P3HT)加入到聚(乙交酯-丙交酯)(PLGA)中,采用静电纺丝工艺,制备了具有电活性和仿生结构的复合纤维。利用酪氨酸羟化酶,将不同质量浓度(10、50、100 ng/mL)的含多巴接头的胰岛素样生长因子-1(DOPA-IGF-1)绑定在纤维表面,实现生长因子长效稳定的作用。通过扫描电子显微镜、接触角表征了纤维直径、分布以及表面亲疏水性。利用细胞培养、荧光染色实验评估了纤维在体外的生物相容性和生物活性。结果表明,该电活性纤维能有效促进大鼠肾上腺嗜铬细胞瘤细胞(PC12)增殖,其中,PLGA/P3HT-5%纤维表现出更好的细胞响应性。结合DOPA-IGF-1质量浓度为10 ng/mL的纤维更利于PC12细胞的黏附、生长。兼具电活性和生物活性的纳米纤维DOPA-IGF-1@PLGA/P3HT在神经组织修复领域具有潜在的应用价值。

关键词: 电活性, 生长因子, 聚噻吩, 聚酯复合物, 静电纺丝, 神经组织工程

Abstract:

An ideal biomaterial for nerve repair should have the properties of similar electrical properties to normal nerve, biomimetic extracellular matrix structure and the release of specific growth factors. In this study, composite fibers with electroactive and bionic structures were prepared by electrospinning process by incorporating various mass fractions(0, 3%, 5%, 10%) poly(3-hexylthiophene)(P3HT) into the poly(glycolide-lactide)(PLGA) polymer. DOPA-jointed insulin-like growth factor-1(DOPA-IGF-1) with different concentrations(10, 50, 100 ng/mL) was bound to the fiber surface using tyrosine hydroxylase to achieve long-term and stable release of growth factor. The fiber diameter and surface hydrophilic properties of the material were characterized by scanning electron microscopy and contact angle measurement, respectively. The biocompatibility and bioactivity of the fibers in vitro were evaluated by cell culture and fluorescence staining experiments. It was found that the electroactive fiber could effectively promote the proliferation of rat adrenal pheochromocytoma cells(PC12 cells), and PLGA/P3HT-5% fiber showed better cell responsiveness. Fibers incorporated with DOPA-IGF-1 with a concentration of 10 ng/mL were more conducive to PC12 cell adhesion and growth. The results showed that DOPA-IGF-1@PLGA/P3HT with both electrical and biological activity had potential application in neural tissue repair.

Key words: electroactive, growth factor, polythiophene, polyester composite, electrospinning, neural tissue engineering

张守燕, 胡江磊, 史新翠, 章培标, 伊藤嘉浩. 电活性和生物活性多巴-胰岛素样生长因子-1@聚(乙交酯-丙交酯)/聚(3-己基噻吩)静电纺丝纤维的制备及神经组织工程应用[J]. 应用化学, 2019, 36(9): 1003-1014.

ZHANG Shouyan, HU Jianglei, SHI Xincui, ZHANG Peibiao, ITO Yoshihiro. Preparation of Electroactive and Bioactive DOPA-Jointed Insulin-Like Growth Factor-1@Poly(glycolide-lactide)/Poly(3-hexylthiophene) Electrospinning Fiber and Its Application in Neural Tissue Engineering[J]. Chinese Journal of Applied Chemistry, 2019, 36(9): 1003-1014.

参考文献

[1]	Wu Y,Wang L,Hu T,et al. Conductive Micropatterned Polyurethane Films as Tissue Engineering Scaffolds for Schwann Cells and PC12 Cells[J]. J Colloid Interface Sci,2018,518:252-262.
[2]	Won S M,Song E,Zhao J,et al. Recent Advances in Materials, Devices, and Systems for Neural Interfaces[J]. Adv Mater,2018,30(30):e1800534.
[3]	Spearman B S,Desai V H,Mobini S,et al. Tissue-Engineered Peripheral Nerve Interfaces[J]. Adv Funct Mater,2017,28(12):1701713.
[4]	Zhu W,Ye T,Lee S J,et al. Enhanced Neural Stem Cell Functions in Conductive Annealed Carbon Nanofibrous Scaffolds with Electrical Stimulation[J]. Nanomed-Nanotechnol,2017,14(7):2485-2494.
[5]	Arioz I,Erol O,Bakan G,et al. Biocompatible Electroactive Tetra(Aniline)-Conjugated Peptide Nanofibers for Neural Differentiation[J]. ACS Appl Mater Interfaces,2018,10(1):308-317.
[6]	Gu X,Ding F,Williams D F.Neural Tissue Engineering Options for Peripheral Nerve Regeneration[J]. Biomaterials,2014,35(24):6143-6156.
[7]	Hsu C C,Serio A,Amdursky N,et al. Fabrication of Hemin-Doped Serum Albumin-Based Fibrous Scaffolds for Neural Tissue Engineering Applications[J]. ACS Appl Mater Interfaces,2018,10(6):5305-5317.
[8]	Jing W,Ao Q,Wang L,et al. Constructing Conductive Conduit with Conductive Fibrous Infilling for Peripheral Nerve Regeneration[J]. Chem Eng J,2018,345:566-577.
[9]	Hajiali H,Contestabile A,Mele E,et al. Influence of Topography of Nanofibrous Scaffolds on Functionality of Engineered Neural Tissue[J]. J Mater Chem B,2018,6(6):930-939.
[10]	Lins L C,Wianny F,Livi S,et al. Effect of Polyvinylidene Fluoride Electrospun Fiber Orientation on Neural Stem Cell Differentiation[J]. J Biomed Mater Res B,2017,105(8):2376-2393.
[11]	Yan L,Zhao B,Liu X,et al. Aligned Nanofibers from Polypyrrole/Graphene as Electrodes for Regeneration of Optic Nerve via Electrical Stimulation[J]. ACS Appl Mater Interfaces,2016,8(11):6834-6840.
[12]	Planellas M,Pérez-Madrigal M M,del Valle L J,et al. Microfibres of Conducting Polythiophene and Biodegradable Poly(ester urea) for Scaffolds[J]. Polym Chem-UK,2015,6(6):925-937.
[13]	He L,Shi Y,Han Q,et al. Surface Modification of Electrospun Nanofibrous Scaffolds via Polysaccharide-Protein Assembly Multilayer for Neurite Outgrowth[J]. J Mater Chem A,2012,22(26):13187-13196.
[14]	Xue J,Yang J,O'Connor D M,et al. Differentiation of Bone Marrow Stem Cells into Schwann Cells for the Promotion of Neurite Outgrowth on Electrospun Fibers[J]. ACS Appl Mater Interfaces,2017,9(14):12299-12310.
[15]	Gopinathan J,Quigley A F,Bhattacharyya A,et al. Preparation, Characterisation, and in Vitro Evaluation of Electrically Conducting Poly(varepsilon-caprolactone)-Based Nanocomposite Scaffolds Using PC12 Cells[J]. J Biomed Mater Res A,2016,104(4):853-865.
[16]	Sun B,Wu T,Wang J,et al. Polypyrrole-Coated Poly(L-Lactic Acid-co-E-Caprolactone)/Silk Fibroin Nanofibrous Membranes Promoting Neural Cell Proliferation and Differentiation with Electrical Stimulation[J]. J Mater Chem B,2016,4(41):6670-6679.
[17]	Yang M,Liang Y,Gui Q,et al. Electroactive Biocompatible Materials for Nerve Cell Stimulation[J]. Mater Res Express,2015,2(4):042001.
[18]	Hardy J G,Lee J Y,Schmidt C E.Biomimetic Conducting Polymer-Based Tissue Scaffolds[J]. Curr Opin Biotechnol,2013,24(5):847-854.
[19]	Swager T M.50th Anniversary Perspective: Conducting/Semiconducting Conjugated Polymers. A Personal Perspective on the Past and the Future[J]. Macromolecules,2017,50(13):4867-4886.
[20]	Mawad D,Artzy-Schnirman A,Tonkin J,et al. Electroconductive Hydrogel Based on Functional Poly(ethylenedioxy Thiophene)[J]. Chem Mater,2016,28(17):6080-6088.
[21]	Koppes A N,Keating K W,McGregor A L,et al. Robust Neurite Extension Following Exogenous Electrical Stimulation Within Single Walled Carbon Nanotube-Composite Hydrogels[J]. Acta Biomater,2016,39:34-43.
[22]	Weng B,Diao J,Xu Q,et al. Bio-interface of Conducting Polymer-Based Materials for Neuroregeneration[J]. Adv Mater Interfaces,2015,2(8):1500059.
[23]	Nguyen H T,Sapp S,Wei C,et al. Electric Field Stimulation Through a Biodegradable Polypyrrole-co-Polycaprolactone Substrate Enhances Neural Cell Growth[J]. J Biomed Mater Res A,2014,102(8) 2554-2564.
[24]	Lin H A,Zhu B,Wu Y W. Dynamic Poly (3,4-Ethylenedioxythiophene)s Integrate Low Impedance with Redox-Switchable Biofunction[J]. Adv Funct Mater,2018,28:1703890.
[25]	Xu H,Holzwarth J M,Yan Y,et al. Conductive Ppy/Pdlla Conduit for Peripheral Nerve Regeneration[J]. Biomaterials,2014,35(1):225-235.
[26]	Shi X,Sui A,Wang Y,et al. Controlled Synthesis of High Molecular Weight Poly(3-Hexylthiophene)s via Kumada Catalyst Transfer Polycondensation with Ni(IPr)(acac)2 as the Catalyst[J]. Chem Commun,2015,51(11):2138-2140.
[27]	Lee J Y,Bashur C A,Goldstein A S,et al. Polypyrrole-Coated Electrospun PLGA Nanofibers for Neural Tissue Applications[J]. Biomaterials,2009,30(26):4325-4335.
[28]	Monkare J,Pontier M,van Kampen E E M,et al. Development of PLGA Nanoparticle Loaded Dissolving Microneedles and Comparison with Hollow Microneedles in Intradermal Vaccine Delivery[J]. Eur J Pharm Biopharm,2018,129:111-121.
[29]	Zhu Y,Wang Z,Zhou H,et al. An Injectable Hydroxyapatite/Poly(Lactide-co-Glycolide) Composite Reinforced by Micro/Nano-Hybrid Poly(glycolide) Fibers for Bone Repair[J]. Mat Sci Eng C-Mater,2017,80:326-334.
[30]	Danhier F,Ansorena E,Silva J M,et al. PLGA-Based Nanoparticles:An Overview of Biomedical Applications[J]. J Control Release,2012,161(2):505-522.
[31]	Quiroga S,Bisbal M,Caceres A.Regulation of Plasma Membrane Expansion During Axon Formation[J]. Dev Neurobiol,2018,78(3):170-180.
[32]	Liang L,Wang J,Yuan Y,et al. Micrna-320 Facilitates the Brain Parenchyma Injury via Regulating IGF-1 During Cerebral I/R Injury in Mice[J]. Bimed Pharmacother,2018,102:86-93.
[33]	Costales J,Kolevzon A.The Therapeutic Potential of Insulin-Like Growth Factor-1 in Central Nervous System Disorders[J]. Neurosci Biobehav R,2016,63:207-222.
[34]	O'Kusky J,Ye P. Neurodevelopmental Effects of Insulin-Like Growth Factor Signaling[J]. Front Neuroendocrin,2012,33(3):230-251.
[35]	Gao T,Zhang N,Wang Z,et al. Biodegradable Microcarriers of Poly(Lactide-co-Glycolide) and Nano-Hydroxyapatite Decorated with IGF-1 via Polydopamine Coating for Enhancing Cell Proliferation and Osteogenic Differentiation[J]. Macromol Biosci,2015,15(8):1070-1080.
[36]	Zhang Y,Wang Z,Wang Y,et al. A Novel Approach via Surface Modification of Degradable Polymers with Adhesive Dopa-IGF-1 for Neural Tissue Engineering[J]. J Pharm Sci-US,2019,108(1):551-562.
[37]	Arasoglu T,Derman S,Mansuroglu B,et al. Synthesis, Characterization and Antibacterial Activity of Juglone Encapsulated PLGA Nanoparticles[J]. J Appl Microbiol,2017,123(6):1407-1419.
[38]	Sun Y,Li H,Lin Y,et al. Integration of Poly(3-Hexylthiophene) Conductive Stripe Patterns with 3D Tubular Structures for Tissue Engineering Applications[J]. RSC Adv,2016,6(76):72519-72524.
[39]	Xu L,Chen S,Lu X,et al. Electrochemically Tunable Cell Adsorption on a Transparent and Adhesion-Switchable Superhydrophobic Polythiophene Film[J]. Macromol Rapid Comm,2015,36(12):1205-1210.
[40]	Li L,Ge J,Wang L,et al. Electroactive Nanofibrous Biomimetic Scaffolds by Thermally Induced Phase Separation[J]. J Mater Chem B,2014,2(36):6119-6130.