电纺壳聚糖-聚乳酸复合神经导管的分子作用及其生物活性表征

杨强; 郭静; 刘树兴; 孙范忱; 陈双

doi:10.13801/j.cnki.fhclxb.20200921.001

电纺壳聚糖-聚乳酸复合神经导管的分子作用及其生物活性表征

doi: 10.13801/j.cnki.fhclxb.20200921.001

杨强^1,,
郭静^{1, 2, ,},
刘树兴^1,,
孙范忱¹,
陈双^1,

1.
大连工业大学　纺织与材料工程学院，大连 116034
2.
辽宁省功能纤维及复合材料工程技术中心，大连 116034

基金项目: 国家自然科学基金(51773024&51373027)；辽宁省科技创新团队项目(LT2017017)

详细信息

通讯作者:
郭静，博士，教授，博士生导师，研究方向为高分子材料改性和纤维材料加工成型　E-mail：guojing8161@163.com

中图分类号: R318.08；TB332
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- 文章访问数: 1155
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- 被引次数: 0
出版历程
- 收稿日期: 2020-07-23
- 录用日期: 2020-09-04
- 网络出版日期: 2020-09-21
- 刊出日期: 2021-06-23

Molecular action and bioactivity characterization of electrospunchitosan-polylactic acid composite nerve conduit

YANG Qiang^1
,,
GUO Jing^{1, 2
, ,},
LIU Shuxing^1
,,
SUN Fanchen¹,
CHEN Shuang^1
,

1.
School of Textile and Engineering, Dalian Polytechnic University, Dalian 116034, China
2.
Functional Fiber and its Composite Materials Engineering Technology Research Center in Liaoning Province, Dalian 116034, China

摘要

摘要: 以三氟乙酸(TFA)、二氯甲烷(DCM)的复合溶液为溶剂，壳聚糖(CS)及聚乳酸(PLA)为溶质制备CS-PLA纺丝液，采用静电纺丝与模板卷取法制备CS-PLA复合神经导管，探究了静电纺CS-PLA复合材料的综合性能。通过表征复合材料的结构与性能，分析了CS与PLA体系内的分子作用，进一步通过复合神经导管与雪旺细胞的共培养实验，提出并验证了雪旺细胞在导管内壁生长黏附时的最佳体系组分。结果表明：复合材料中CS与PLA分子间以氢键结合，其中各种氢键贡献作用大小依次是OH…OH型>OH…N型>OH…醚O型。PLA含量增加，复合材料的热性能和热稳定性提高，断裂强度提升197.7%，断裂伸长率提高53.7%。CS含量增加，复合材料的亲水性和细胞相容性逐渐增加。在组分比例CS∶PLA(3∶1)时，凹槽及微孔的大小最有利于雪旺细胞的黏附。
- 复合神经导管 /
- 静电纺丝 /
- 聚乳酸 /
- 壳聚糖 /
- 氢键作用
Abstract: The chitosan (CS)-polylactic acid (PLA) spinning solution was prepared using a composite solution of trifluoroacetic acid (TFA) and dichloromethane (DCM) as the solvent, and CS and PLA as the solute. Electrospinning and template coiling were used to prepare CS-PLA composite nerve conduit, and the comprehensive properties of CS-PLA composites were investigated. By characterizing the structure and properties of composites, the molecular interactions in CS and PLA systems were analyzed. Furthermore, through the co-cultivation experiments of composite nerve conduit and Schwann cells, the best system components for Schwann cells to grow and adhere to the inner wall of the catheter were proposed and verified. The results show that CS and PLA molecules are bonded by hydrogen bonds in the composite material, and the various hydrogen bond contributions are in the order of OH...OH>OH...N>OH...ether O. With the increase of PLA content, the thermal performance and thermal stability of the composites are increased, the tensile strength is increased by 197.7%, and the breaking elongation is increased by 53.7%. With the increase of CS content, the hydrophilicity and cell compatibility of the composites gradually increase. When the component ratio of CS∶PLA is 3∶1, the size of the grooves and micropores is the most favorable for Schwann cell adhesion.
- composite nerve conduit /
- electrospinning /
- polylactic acid /
- chitosan /
- hydrogen bonding

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图 1 壳聚糖(CS)-聚乳酸(PLA)复合神经导管的制备过程

Figure 1. Preparation of chitosan (CS)-polylactic acid (PLA) composite nerve conduit

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图 2 CS、PLA及CS-PLA复合材料的FTIR图谱 (a)、不同比例的CS-PLA复合材料吸光度对比图 (b)

Figure 2. FTIR spectra of CS, PLA and CS-PLA composites (a), comparison diagram of absorbance of CS-PLA composites in different ratios (b)

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图 3 CS-PLA 复合材料的高斯分峰拟合及各子峰的分布

Figure 3. Gaussian peak fitting of CS-PLA composites and distribution of each subpeak ((a) CS∶PLA (2∶1); (b) CS∶PLA (3∶1); (c) CS∶PLA (4∶1))

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图 4 CS与PLA氢键作用过程

Figure 4. Mechanism of hydrogen bonding between CS and PLA

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图 5 CS、PLA及各组分CS-PLA复合材料的DTG (a) 与TG (b) 曲线

Figure 5. TGA curves (a) and DTG curves (b) of CS, PLA and CS-PLA composites

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图 6 CS、PLA及CS-PLA复合神经导管的DSC升温曲线

Figure 6. DSC heating curves of CS, PLA and CS-PLA composite nerve conduit

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图 7 CS及CS-PLA复合材料的H₂O迁移示意图

Figure 7. H₂O migration diagram of crystallization of CS-PLA composites

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图 8 CS、PLA(a)及CS-PLA复合材料(b)的XRD图谱

Figure 8. XRD patterns of CS, PLA (a) and CS-PLA composites (b)

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图 9 CS-PLA复合神经导管内壁的SEM图像

Figure 9. SEM images of the inner wall of CS-PLA composite nerve conduit((a) CS∶PLA (4∶1); (b) CS∶PLA (3∶1); (c) CS∶PLA (2∶1))

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图 10 CS-PLA复合材料的接触角测试图

Figure 10. Contact angle test pictures of CS-PLA composites ((a) CS∶PLA (2∶1); (b) CS∶PLA (3∶1); (c)CS∶PLA (4∶1))

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图 11 CS-PLA复合神经导管体外细胞培养的SEM图像

Figure 11. SEM images of in vitro cell culture of CS-PLA composite nerve conduit ((a) CS∶PLA (2∶1); (b) CS∶PLA (3∶1); (c)CS∶PLA (4∶1))

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表 1 各种氢键类型的拟合结果

Table 1. Fitting results of various kinds of hydrogen bonds

Mass ratio	Hydrogen bond type		Abbreviation	Wavenumber/ cm⁻¹	Relative strength/%	Total relative strength/%
CS∶PLA (2∶1)	Free hydroxyl	V	—OH	3563	14.0	14.0
	Intramolecular hydrogen bond	Ⅲ	OH…OH	3405	35.9	70.3
		I	Annular polymer	3126	2.7
		IV	OH…N	3516	31.7
	Intermolecular hydrogen bond	II	OH…ether O	3250	15.7	15.7
CS∶PLA (3∶1)	Free hydroxyl	V	—OH	3593	10.7	10.7
	Intramolecular hydrogen bond	Ⅲ	OH…OH	3403	42.8	72.0
		I	Annular polymer	3112	1.9
		IV	OH…N	3502	27.3
	Intermolecular hydrogen bond	II	OH…ether O	3244	17.3	17.3
CS∶PLA (4∶1)	Free hydroxyl	V	—OH	3558	7.6	7.6
	Intramolecular hydrogen bond	Ⅲ	OH…OH	3405	38.1	74.5
		I	Annular polymer	3132	2.7
		IV	OH…N	3508	33.7
	Intermolecular hydrogen bond	II	OH…ether O	3259	17.9	17.9

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表 2 CS-PLA复合材料的断裂强度和断裂伸长率

Table 2. Tensile strength and breaking elongation of CS-PLA composites

CS∶PLA (Mass ratio)	Tensile strength/MPa	Breaking elongation/%
4∶1	0.351	9.09
3∶1	0.585	10.91
2∶1	1.045	13.79

下载: 导出CSV

参考文献(30)

[1]	林彦杰, 高宝云, 李勇峰. 组织工程材料在周围神经损伤修复中应用的研究进展[J]. 中国医学工程, 2018, 26(1):20-22. LIN Yanjie, GAO Baoyun, LI Yongfeng. Research progress of tissue engineering materials in the repair of peripheral nerve injury[J]. China Medical Engineering,2018,26(1):20-22(in Chinese).
[2]	LÓPEZ-CEBRAL R, SILVA-CORREIA J, REIS R L, et al. Peripheral nerve injury: Current challenges, conventional treatment approaches, and new trends in biomaterials-based regenerative strategies[J]. ACS Biomaterials Science & Engineering,2017,3(12):3098-3122.
[3]	ABALYMOV A, PARAKHONSKIY B, SKIRTACH A G. Polymer- and hybrid-based biomaterials for interstitial, connective, vascular, nerve, visceral and musculoskeletal tissue engineering[J]. Polymers (Basel),2020,12(3):620.
[4]	BADEA S, WU W. Nanoengineered biomaterials for bridging gaps in damaged nerve tissue[J]. Nanoengineered Biomaterials for Regenerative Medicine,2019:187-214.
[5]	JAHROMI M, RAZAVI S, BAKHTIARI A. The advances in nerve tissue engineering: From fabrication of nerve conduit to in vivo nerve regeneration assays[J]. Journal of Tissue Engineering and Regenerative,2019,13(11):2077-2100.
[6]	张会兰, 易兵成, 王先流, 等. 用高度取向石墨烯/聚乳酸(Gr/PLLA)复合超细纤维构建神经导管[J]. 高等学校化学学报, 2016, 37(5):972-982. ZHANG Huilan, YI Bingcheng, WANG Xianliu, et al. Constructing neural catheters with highly oriented graphene/polylactic acid (Gr/PLLA) composite ultrafine fibers[J]. Chemical Journal of Chinese Universities,2016,37(5):972-982(in Chinese).
[7]	PHILIBERT T, LEE B H, FABIEN N. Current status and new perspectives on chitin and chitosan as functional biopolymers[J]. Appl Biochem Biotechnol,2017,181(4):1314-1337.
[8]	李高荣, 欧阳茜茜, 李思东, 等. 壳聚糖纳米粒子的制备及其在医药领域的应用[J]. 药学研究, 2018, 37(1):53-56. LI Gaorong, OUYANG Qianqian, LI Sidong, et al. Preparation of chitosan nanoparticles and their application in the field of medicine[J]. Pharmaceutical Research,2018,37(1):53-56(in Chinese).
[9]	陈潇, 张浩宇, 霍神焕, 等. 壳聚糖改性地聚合物的力学及吸附性能[J]. 复合材料学报, 2019, 36(12):2959-2967. CHEN Xiao, ZHANG Haoyu, HUO Shenhuan, et al. Mechanical and adsorption properties of chitosan-modified geopolymers[J]. Journal of Composite Materials,2019,36(12):2959-2967(in Chinese).
[10]	YE H, ZHU J, DENG D, et al. Enhanced osteogenesis and angiogenesis by PCL/chitosan/Sr-doped calcium phosphate electrospun nanocomposite membrane for guided bone regeneration[J]. Journal of Biomaterials Science, Polymer Edition,2019,30(16):1505-1522.
[11]	王栋, 宣丽慧, 李超, 等. 静电纺纤维素纳米晶体/壳聚糖-聚乙烯醇复合纳米纤维的制备与表征[J]. 复合材料学报, 2018, 35(4):964-972. WANG Dong, XUAN Lihui, LI Chao, et al. Preparation and characterization of electrospun cellulose nanocrystals/chitosan-polyvinyl alcohol composite nanofibers[J]. Acta Materiae Compositae Sinica,2018,35(4):964-972(in Chinese).
[12]	詹世平, 万泽韬, 王景昌, 等. 生物医用材料聚乳酸的合成及其改性研究进展[J]. 化工进展, 2020, 39(1):199-205. ZHAN Shiping, WAN Zetao, WANG Jingchang, et al. Progress in the synthesis and modification of biomedical materials polylactic acid[J]. Progress in Chemical Engineering,2020,39(1):199-205(in Chinese).
[13]	MISHRA R K, HA S K, VERMA K, et al. Recent progress in selected bio-nanomaterials and their engineering applications: An overview[J]. Journal of Science: Advanced Materials and Devices,2018,3(3):263-288.
[14]	陈立红, 雷志敏, 王莉莉. 碱性成纤维细胞生长因子/壳聚糖/聚乳酸支架用于牙周组织再生工程的体外实验[J]. 中国组织工程研究, 2017, 21(26):4106-4112. doi: 10.3969/j.issn.2095-4344.2017.26.002 CHEN Lihong, LEI Zhimin, WANG Lili. In vitro experiment of basic fibroblast growth factor/chitosan/polylactic acid scaffold for periodontal tissue regeneration engineer-ing[J]. China Tissue Engineering Research,2017,21(26):4106-4112(in Chinese). doi: 10.3969/j.issn.2095-4344.2017.26.002
[15]	KASIRAJAN S, UMAPATHY D, CHANDRASEKAR C, et al. Preparation of poly(lactic acid) from Prosopis juliflora and incorporation of chitosan for packaging applications[J]. Journal of Bioscience and Bioengineering,2019,128(3):323-331.
[16]	SUN, F, GUO, J, LIU, Y, et al. Preparation and characterization of poly(3-hydroxybutyrate-co-4-hydroxybutyrate)/pullulan-gelatin electrospun nanofibers with shell-core structure[J]. Biomedical Materials,2020,15(4):045023.
[17]	PARHAM S, KHARAZI A Z, BAKHSHESHI-RAD H R, et al. Electrospun nano-fibers for biomedical and tissue en-gineering applications: A comprehensive review[J]. Materials (Basel),2020,13(9):2153.
[18]	蒋岩岩, 秦静雯, 王鸿博. 壳聚糖/聚乳酸复合纳米纤维的制备及抗菌性能研究[J]. 材料导报, 2012, 26(18):74-76, 80. doi: 10.3969/j.issn.1005-023X.2012.18.020 JIANG Yanyan, QIN Jingwen, WANG Hongbo. Preparation and antibacterial properties of chitosan/polylactic acid composite nanofibers[J]. Materials Herald,2012,26(18):74-76, 80(in Chinese). doi: 10.3969/j.issn.1005-023X.2012.18.020
[19]	AU H T, PHAM L N, VU T H T, et al. Fabrication of an antibacterial non-woven mat of a poly(lactic acid)/chitosan blend by electrospinning[J]. Macromolecular Research,2011,20(1):51-58.
[20]	NGUYEN T T T, CHUNG O H, PARK J S. Coaxial electrospun poly (lactic acid)/chitosan (core/shell) composite nanofibers and their antibacterial activity[J]. Carbohydrate Polymers,2011,86(4):1799-1806.
[21]	王华林, 丁曼, 翟林峰, 等. 壳聚糖/聚乳酸复合纳米纤维的制备与表征[J]. 高分子材料科学与工程, 2013, 29(1):162-165. WANG Hualin, DING Man, ZHAI Linfeng, et al. Preparation and characterization of chitosan/polylactic acid composite nanofibers[J]. Polymer Materials Science and Engineering,2013,29(1):162-165(in Chinese).
[22]	XU J, ZHANG J, GAO W, et al. Preparation of chitosan/PLA blend micro/nanofibers by electrospinning[J]. Materials Letters,2009,63(8):658-660.
[23]	HU Y, WU X, JINRUI X. Self-assembled supramolecular hydrogels formed by biodegradable PLA/CS diblock copolymers and beta-cyclodextrin for controlled dual drug delivery[J]. International Journal of Biological Macromolecules: Structure, Function and Interactions,2018,108:18-23.
[24]	李婷, 刘君泰. Origin软件的高斯多峰拟合方法在物理化学实验中的应用[J]. 化工设计通讯, 2016, 42(5):157. doi: 10.3969/j.issn.1003-6490.2016.05.125 LI Ting, LIU Juntai. Application of the Gaussian multi-peak fitting method of origin software in physical chemistry experiments[J]. Chemical Engineering Design Newsletter,2016,42(5):157(in Chinese). doi: 10.3969/j.issn.1003-6490.2016.05.125
[25]	CHEN J, GUO J, ZHAO M, et al. Hydrogen bonding in chitosan/Antarctic krill protein composite system: Study on construction and enhancement mechanism[J]. International Journal of Biological Macromolecules,2020:142, 513-520.
[26]	WANG Y, CHANG Y, YU L, et al. Crystalline structure and thermal property characterization of chitin from Antarctic krill (Euphausia superba)[J]. Carbohydr Polym,2013,92(1):90-97.
[27]	PAL A K, KATIYAR V. Thermal degradation behaviour of nanoamphiphilic chitosan dispersed poly (lactic acid) bionanocomposite films[J]. International Journal of Biological Macromolecules,2017,95:1267-1279.
[28]	琚海燕, 但卫华, 但年华, 等. 壳聚糖/聚己内酯复合膜的微观形态与结构特征[J]. 复合材料学报, 2016, 33(9):2038-2044. JU Haiyan, DAN Weihua, DAN Nianhua, et al. Micromorphology and structural characteristics of chitosan/polycaprolactone composite membrane[J]. Acta Materiae Compositae Sinica,2016,33(9):2038-2044(in Chinese).
[29]	雷雁洲, 王少伟, 吕秦牛, 等. 碳纤维/聚乳酸复合材料的结晶性能和流变特性[J]. 复合材料学报, 2018, 35(6):1402-1406. LEI Yanzhou, WANG Shaowei, LV Qinniu, et al. Crystallization and rheological properties of carbon fiber/polylactic acid composites[J]. Acta Materiae Compositae Sinica,2018,35(6):1402-1406(in Chinese).
[30]	JENKINS P M, LAUGHTER M R, LEE D J, et al. A nerve guidance conduit with topographical and biochemical cues: Potential application using human neural stem cells[J]. Nanoscale Research Letters,2015,10(1):972.