丝素蛋白与聚丁二酸丁二醇酯复合超细纤维膜的制备及性能研究
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摘要
本文旨在改善静电纺丝素蛋白(SF)超细纤维膜力学性能的不足,通过共混、混杂和同轴静电纺丝制备出结构不同的SF/PBS复合超细纤维膜,以期在提高其力学性能的同时,通过生物学评价可以获得具有良好细胞相容性的SF/PBS组织工程支架,以扩大静电纺丝素蛋白在组织工程支架材料领域的应用。
首先研究了静电纺PBS的纺丝工艺,当质量分数为6%、电场强度为1.5kv/cm(纺丝电压18kv,纺丝距离12cm)和纺丝流率为0.01ml/min时,所得到的静电纺PBS超细纤维的形貌较好。通过对比分析静电纺PBS超细纤维膜与浇铸膜的结构及力学性能,静电纺PBS超细纤维膜结晶性能降低;拉伸破坏强度有所下降,为18.6MPa,但破坏应变明显增大,达到120%,表现出良好的力学性能。
然后通过共混和混杂静电纺丝分别制备了结构不同的SF/PBS复合超细纤维膜,研究了甲醇处理前后的形貌、结构和力学性能。并进一步研究了PBS(核层)和SF(壳层)的同轴静电纺丝,分析了核壳超细纤维膜甲醇处理前后的结构及力学性能。最后对利用共混、混杂和同轴静电纺丝所制备的SF/PBS复合超细纤维膜进行生物学评价,研究了不同支架材料的细胞相容性。结论如下:
(1)共混复合超细纤维膜经甲醇处理后的结晶性能随着PBS质量比的增加,结晶性能变好,相同共混质量比的复合超细纤维膜处理后结晶性变好;共混复合超细纤维膜处理后的拉伸破坏应力随着PBS质量比的增加,拉伸破坏应力先减小后增大,应变逐渐增大;相同共混质量比的复合超细纤维膜处理后的拉伸破坏应力变大,应变减小,当共混质量比为50/50时,应力、应变分别达到16MPa和50%,复合超细纤维膜表现出较好的力学性能。
(2)混杂复合超细纤维膜的结晶性能经甲醇处理后随着PBS质量比的增加,结晶性能变好,相同混杂质量比的复合超细纤维膜经甲醇处理后结晶性能变好。甲醇处理后混杂复合超细纤维膜随着PBS含量的增加,拉伸破坏应力逐渐增大,应变也逐渐增大;相同混杂比的复合超细纤维膜经甲醇处理后的拉伸破坏应力增加,但应变减小。
(3)通过FESEM和TEM的表征,成功制备了静电纺PBS/SF核壳结构超细纤维,经甲醇处理后由于壳层厚度的增加导致核层的衍射吸收强度减小,表现为整个超细复合纤维膜的结晶性能下降。与未经甲醇处理相比,处理后的静电纺PBS/SF核壳结构超细纤维膜的拉伸破坏应力从14.9MPa增大到17.2MPa,但是拉伸破坏应变从96.8%减小到81.8%。
(4)通过L929细胞黏附和增殖实验,静电纺SF/PBS共混、混杂和同轴核壳结构超细纤维支架在细胞黏附上没有明显区别。静电纺SF/PBS复合超细纤维支架在细胞增殖上有一定的优势,尤其是SF/PBS共混质量比为50/50和PBS/SF核壳结构的支架材料有明显优势,其OD值已高于纯丝素蛋白的OD值,再结合力学性能可知,这两种支架材料在组织工程支架领域的应用有较大的潜力。
The SF/PBS ultrafine composite fibrous membranes of different structures were fabricated by the blend, hybrid and coaxial electrospinning in order to synchronously enhance the mechanical property of SF, and also obtain SF/PBS composite tissue engineering scaffolds, which possess a better cytocompatibility depending on biological evaluation, and finally further extend the application range of SF in biomaterials.
To prepare SF/PBS ultrafine composite fibrous membranes, the parameters in electrospinning PBS process were studied. the morphology of the received electrospun PBS ultrafine fibers is better, when the mass fraction of 6%, electric field strength 1.5kv/cm (spinning voltage of 18kv, spinning distance of 12cm) and spinning flow rate of 0.01ml/min. Comparison with the PBS cast membrane, the crystallinity of electrospun PBS ultrafine fibrous membranes decreases, the tensile failure strength slightly decreases to 18.6MPa, but failure elongation is almost the double of cast membranes up to 120%.
The SF/PBS ultrafine composite fibrous membranes of different structure are prepared by the blend and hybrid electrospinning, and the morphology, structure and mechanical properties are also studied before and after treatment of methanol. Coaxial electrospinning of PBS (core layer) and SF (shell layer) was also further studied. The morphology, structure and mechanical properties of PBS/SF core/shell ultrafine fibrous membrane were also tested. Finally, Cell biocompatibility was evaluated on different SF/PBS ultrafine composite fibrous scaffolds prepared by the blend, hybrid and coaxial electrospinning. The main results of this work are summarized as follow:
(1) After treatment with methanol, the crystallization properties of electrospun SF/PBS blend ultrafine composite fibrous membranes gradually becomes better with addition of PBS, and the crystallization properties on the ultrafine composite fibous membranes with same mass ratio is better. The tensile strength first decreases and then increases with addition of PBS, and failure strain gradually increases. For the same blend mass ratio, failure strength increases, but failure strain decreases. When the blend ratio is 50/50, failure strength is close to 16MPa and failure strain is up to 50%, so mechanical properties of SF/PBS blend ultrafine fibrous membranes was improved.
(2) The crystallization properties of the whole hybrid composite ultrafine fibrous membranes treated with methanol also increase with addition of PBS, and the crystallization properties on the same hybrid mass ratio is better than that before treatment. in addition, the tensile strength gradually increases with addition of PBS, and also failure strain gradually increases. For the same hybrid mass ratio, tensile strength increase, but failure strain decreases. When the hybrid mass ratio is 50/50, mechanical properties of SF/PBS hybrid ultrafine fibrous membrane was best, failure strength was up to 17.3MPa and failure strain was 38.2%.
(3) The PBS/SF core/shell ultrafine fibers was successfully prepared by coaxial electrospinning, After treatment with methanol, due to the increasing of shell thickness, absorption intensity of X-ray diffraction in core PBS decreases, which lead to the whole crystallization properties of PBS/SF core/shell ultrafine fibrous membranes dropped. Compared with non-treatment, the electrospun PBS/SF core/shell ultrafine fibrous membrane tensile strength increased from 14.9MPa to 17.2MPa, but the failure strain decreased from 96.8% to 81.8 %.
(4) Through L929 cell adhesion experiment, it was found that the blend, hybrid, and core-shell ultrafine composite fibrous scaffolds do not have significant difference. However, for proliferation experiment, they have certain advantages, especially scaffolds of blend mass ratio 50/50 and core/shell scaffolds show more obvious advantages, their OD values are higher than that of SF, and the mechanical properties are better, so we can see that these scaffolds have great application potential in tissue engineering scaffolds.
首先研究了静电纺PBS的纺丝工艺,当质量分数为6%、电场强度为1.5kv/cm(纺丝电压18kv,纺丝距离12cm)和纺丝流率为0.01ml/min时,所得到的静电纺PBS超细纤维的形貌较好。通过对比分析静电纺PBS超细纤维膜与浇铸膜的结构及力学性能,静电纺PBS超细纤维膜结晶性能降低;拉伸破坏强度有所下降,为18.6MPa,但破坏应变明显增大,达到120%,表现出良好的力学性能。
然后通过共混和混杂静电纺丝分别制备了结构不同的SF/PBS复合超细纤维膜,研究了甲醇处理前后的形貌、结构和力学性能。并进一步研究了PBS(核层)和SF(壳层)的同轴静电纺丝,分析了核壳超细纤维膜甲醇处理前后的结构及力学性能。最后对利用共混、混杂和同轴静电纺丝所制备的SF/PBS复合超细纤维膜进行生物学评价,研究了不同支架材料的细胞相容性。结论如下:
(1)共混复合超细纤维膜经甲醇处理后的结晶性能随着PBS质量比的增加,结晶性能变好,相同共混质量比的复合超细纤维膜处理后结晶性变好;共混复合超细纤维膜处理后的拉伸破坏应力随着PBS质量比的增加,拉伸破坏应力先减小后增大,应变逐渐增大;相同共混质量比的复合超细纤维膜处理后的拉伸破坏应力变大,应变减小,当共混质量比为50/50时,应力、应变分别达到16MPa和50%,复合超细纤维膜表现出较好的力学性能。
(2)混杂复合超细纤维膜的结晶性能经甲醇处理后随着PBS质量比的增加,结晶性能变好,相同混杂质量比的复合超细纤维膜经甲醇处理后结晶性能变好。甲醇处理后混杂复合超细纤维膜随着PBS含量的增加,拉伸破坏应力逐渐增大,应变也逐渐增大;相同混杂比的复合超细纤维膜经甲醇处理后的拉伸破坏应力增加,但应变减小。
(3)通过FESEM和TEM的表征,成功制备了静电纺PBS/SF核壳结构超细纤维,经甲醇处理后由于壳层厚度的增加导致核层的衍射吸收强度减小,表现为整个超细复合纤维膜的结晶性能下降。与未经甲醇处理相比,处理后的静电纺PBS/SF核壳结构超细纤维膜的拉伸破坏应力从14.9MPa增大到17.2MPa,但是拉伸破坏应变从96.8%减小到81.8%。
(4)通过L929细胞黏附和增殖实验,静电纺SF/PBS共混、混杂和同轴核壳结构超细纤维支架在细胞黏附上没有明显区别。静电纺SF/PBS复合超细纤维支架在细胞增殖上有一定的优势,尤其是SF/PBS共混质量比为50/50和PBS/SF核壳结构的支架材料有明显优势,其OD值已高于纯丝素蛋白的OD值,再结合力学性能可知,这两种支架材料在组织工程支架领域的应用有较大的潜力。
The SF/PBS ultrafine composite fibrous membranes of different structures were fabricated by the blend, hybrid and coaxial electrospinning in order to synchronously enhance the mechanical property of SF, and also obtain SF/PBS composite tissue engineering scaffolds, which possess a better cytocompatibility depending on biological evaluation, and finally further extend the application range of SF in biomaterials.
To prepare SF/PBS ultrafine composite fibrous membranes, the parameters in electrospinning PBS process were studied. the morphology of the received electrospun PBS ultrafine fibers is better, when the mass fraction of 6%, electric field strength 1.5kv/cm (spinning voltage of 18kv, spinning distance of 12cm) and spinning flow rate of 0.01ml/min. Comparison with the PBS cast membrane, the crystallinity of electrospun PBS ultrafine fibrous membranes decreases, the tensile failure strength slightly decreases to 18.6MPa, but failure elongation is almost the double of cast membranes up to 120%.
The SF/PBS ultrafine composite fibrous membranes of different structure are prepared by the blend and hybrid electrospinning, and the morphology, structure and mechanical properties are also studied before and after treatment of methanol. Coaxial electrospinning of PBS (core layer) and SF (shell layer) was also further studied. The morphology, structure and mechanical properties of PBS/SF core/shell ultrafine fibrous membrane were also tested. Finally, Cell biocompatibility was evaluated on different SF/PBS ultrafine composite fibrous scaffolds prepared by the blend, hybrid and coaxial electrospinning. The main results of this work are summarized as follow:
(1) After treatment with methanol, the crystallization properties of electrospun SF/PBS blend ultrafine composite fibrous membranes gradually becomes better with addition of PBS, and the crystallization properties on the ultrafine composite fibous membranes with same mass ratio is better. The tensile strength first decreases and then increases with addition of PBS, and failure strain gradually increases. For the same blend mass ratio, failure strength increases, but failure strain decreases. When the blend ratio is 50/50, failure strength is close to 16MPa and failure strain is up to 50%, so mechanical properties of SF/PBS blend ultrafine fibrous membranes was improved.
(2) The crystallization properties of the whole hybrid composite ultrafine fibrous membranes treated with methanol also increase with addition of PBS, and the crystallization properties on the same hybrid mass ratio is better than that before treatment. in addition, the tensile strength gradually increases with addition of PBS, and also failure strain gradually increases. For the same hybrid mass ratio, tensile strength increase, but failure strain decreases. When the hybrid mass ratio is 50/50, mechanical properties of SF/PBS hybrid ultrafine fibrous membrane was best, failure strength was up to 17.3MPa and failure strain was 38.2%.
(3) The PBS/SF core/shell ultrafine fibers was successfully prepared by coaxial electrospinning, After treatment with methanol, due to the increasing of shell thickness, absorption intensity of X-ray diffraction in core PBS decreases, which lead to the whole crystallization properties of PBS/SF core/shell ultrafine fibrous membranes dropped. Compared with non-treatment, the electrospun PBS/SF core/shell ultrafine fibrous membrane tensile strength increased from 14.9MPa to 17.2MPa, but the failure strain decreased from 96.8% to 81.8 %.
(4) Through L929 cell adhesion experiment, it was found that the blend, hybrid, and core-shell ultrafine composite fibrous scaffolds do not have significant difference. However, for proliferation experiment, they have certain advantages, especially scaffolds of blend mass ratio 50/50 and core/shell scaffolds show more obvious advantages, their OD values are higher than that of SF, and the mechanical properties are better, so we can see that these scaffolds have great application potential in tissue engineering scaffolds.
引文
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