Preparation and biocompatibility of EDC-NHS crosslinked chitosan-polyethyleneoxide-silk fibroin electrospun nanofibers
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摘要: 为提高壳聚糖(CS)-丝素蛋白(SF)复合纳米纤维的可纺性,解决其易溶胀从而导致纳米纤维尺寸稳定性较差的问题,采用聚氧化乙烯(PEO)提高纳米纤维的可纺性,用静电纺丝来制备纳米纤维,然后通过1-(3-二甲基氨基丙基)-3-乙基碳化二亚胺(EDC)和 N-羟基丁二酰亚胺(NHS)进行交联改性,制备了CS-PEO-SF三元体系的纳米纤维。利用FTIR、XRD、单纤维强力机、SEM分别表征了CS-PEO-SF纳米纤维的分子间相互作用、结晶性能、纳米纤维的力学性能和形貌,同时考察纳米纤维的溶胀度;利用细胞毒性和细胞培养测试表征了CS-PEO-SF纳米纤维的生物相容性。结果表明:PEO的加入可以有效提高CS-SF的可纺性,CS-PEO-SF纳米纤维的平均直径范围为240~510 nm,各组分纤维形态较好;随着SF含量的增加,CS-PEO-SF纳米纤维结晶性能、拉伸强度和断裂伸长率逐渐增大。经交联后的CS-PEO-SF纳米纤维结晶能力和溶胀度降低,力学性能提高。交联前后CS-PEO-SF纳米纤维均不具毒性,有良好的细胞相容性。Abstract: In order to improve the spinnability of chitosan (CS)-silk fibroin (SF) composite nanofibers and solve the problem of poor dimensional stability of nanofiber membrane caused by its easy swelling, polyethylene oxide (PEO) was used to improve the spinnability of the composite fibers in this study. Electrospinning was used to prepare CS-PEO-SF nanofibers, which were cross-linked and by 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) and N-hydroxysuccinimide (NHS). The intermolecular interaction, crystallization properties, mechanical properties and morphology of CS-PEO-SF nanofibers were characterized by FTIR, XRD, single fiber strength machine and SEM. The swelling degree and biocompatibility of CS-PEO-SF nanofibers was also characterized by cytotoxicity and cell culture tests. The results show that the addition of PEO can effectively improve the spinnability of CS-SF. The average diameter of CS-PEO-SF nanofibers is from 240 nm to 510 nm. The fiber morphology of each component is good. With the increase of SF content, the crystalline properties, tensile strength and breaking elongation of CS-PEO-SF nanofibers gradually increase. After crosslinking, the crystallinity and swelling degree of the CS-PEO-SF nanofibers decrease, and the mechanical properties increase. Before and after cross-linking, the CS-PEO-SF nanofibers has no toxicity and has good cell compatibility.
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Key words:
- chitosan /
- polyethylene oxide /
- silk fibroin /
- nanofibers /
- cross-linking /
- biocompatibility
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表 1 纺丝液各成分的质量比
Table 1. Mass ratios of each components of spinning solution
Smaple CS PEO SF CS-PEO-SF1 10 1 0 CS-PEO-SF2 10 1 1 CS-PEO-SF3 10 1 1.5 CS-PEO-SF4 10 1 2 CS-PEO-SF5 10 1 2.5 CS-PEO-SF6 10 1 3 CS-PEO-SF7 10 1 3 Notes: CS—Chitosan; PEO—Polyethylene oxide; SF—Silk fibroin; CS-PEO-SF6—CS-PEO-SF nanofiber before crosslinking; CS-PEO-SF7—Cross-linked CS-PEO-SF nanofiber. 表 2 细胞增殖率与细胞毒性水平的对应关系
Table 2. Correspondence between cell proliferation rate and cytotoxicity level
Cell proliferation rate/% Cytotoxicity level ≥100 0 75–99 1 50–74 2 25–49 3 1–24 4 0 5 表 3 CS-PEO-SF纳米纤维的平均直径
Table 3. Average diameters of CS-PEO-SF nanofibers
Mass ratio of CS to SF Average diameter/μm 10∶0 0.24 10∶1.0 0.25 10∶1.5 0.34 10∶2.0 0.42 10∶2.5 0.45 10∶3.0 0.51 -
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