疏水蛋白对静电纺PLGA支架和PLGA膜进行表面修饰及其应用的研究
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摘要
生物多聚物Poly(D,L-lactic-co-glycolic acid)polymers(PLGA)具有良好的生物相容性和生物可降解性,是目前组织工程研究中使用较多的支架材料。但PLGA存在亲水性差、细胞粘附力弱等问题,限制了其的应用。因此,开发一种能够有效改善PLGA表面特性的方法成为当前研究的热点。
本文引进了一种Ⅱ型疏水蛋白HFBI来修饰PLGA,并进一步研究了HFBI修饰后的PLGA在作为细胞培养基底及制备图案化神经干细胞复合支架材料方面的应用问题。
疏水蛋白是一种从丝状真菌中提取的具有两亲性及表面活性的蛋白质,且已被证实能够改变疏水材料的亲/疏水性。通过测量HFBI修饰后的静电纺PLGA支架和PLGA膜表面的静态疏水角,发现二者的表面亲水性及湿润性都显著提高了。在此基础上对静电纺PLGA支架进一步进行胶原修饰并得到其扫描电镜图像,发现胶原能够很好的贴附到HFBI修饰后的PLGA支架上。通过在HFBI/胶原共修饰的静电纺PLGA支架上培养细胞,发现细胞在支架上贴附生长状态良好。该结果不仅解决了PLGA材料细胞贴附率低的问题,还为利用PLGA膜制备图案化神经干细胞复合支架器件奠定了基础。因此,利用微接触印刷法将血清印迹到HFBI修饰的PLGA膜上,并在此基底上培养神经干细胞,成功地获得了图案化神经干细胞复合支架。利用HFBI来修饰静电纺PLGA支架和PLGA膜,不仅能有效的改善表/界面材料的亲疏水性,为生物分子、细胞粘附和生长提供便利,还进一步拓展了PLGA在组织工程中的应用。
Biopolymer poly(lacic-co-glycolic acid) PLGA has been extensively applied in tissue engineering as the scaffold materials, due to its good biocompatibility and biodegradability. However, the hydrophobility property of the biopolymer PLGA materials gives rise to the poor cell affinity on the surface and limits their application. Therefore, it is a hot topic to develop an efficient approach to increasing the hydrophilicity of PLGA materials.
In this study, we introduced a class of hydrophobin to modify the biopolymer PLGA. Besides, we also investigated its application both in cell adhesion and the preparation of neural stem cells (NSCs) patterns which made the basis for the fabrication of the complex scaffold composed of NSCs and PLGA film.
Hydrophobins are a group of amphiphilic and surface-active proteins found in filamentous fungi, which has been used to change the surface property of hydrophobic materials. The classⅡhydrophobin HFBI was used to modify electrospun PLGA scaffold and PGLA film. The static water contact angle of the HFBI-modified electrospun PLGA scaffold and PLGA film showed that their surface property conversed from hydrophobictiy to hydrophilicity. Collagen was modified on the HFBI-pretreated electrospun scaffold and the SEM image of the HFBI/collagen co-coated PLGA scaffold demonstrated that collagen could immobilized on the PLGA scaffold with high efficiency. When the HFBI/collagen co-coated PLGA scaffold was used as the substrate of culturing 293 T cells, the cells adhered and grew well on the surface. This result overcame the disadvantage of PLGA materials mentioned above and established the possibility of fabricating neural stem cells patterns on the PLGA film as well. Therefore, the micro-contact printing technology was used to prepare the serum patterns on the HFBI-modified PLGA film and the well-defined neural stem cells patterns was obtained by directly culturing cells on the PLGA film.
HFBI modification of electrospun PLGA scaffold and PLGA film could not only efficiently change the hydrophobicity surface property and facilitate both the immoblization of biomolecular and cell adhesion on the surface, but also enlarge their application in the tissue engineering.
本文引进了一种Ⅱ型疏水蛋白HFBI来修饰PLGA,并进一步研究了HFBI修饰后的PLGA在作为细胞培养基底及制备图案化神经干细胞复合支架材料方面的应用问题。
疏水蛋白是一种从丝状真菌中提取的具有两亲性及表面活性的蛋白质,且已被证实能够改变疏水材料的亲/疏水性。通过测量HFBI修饰后的静电纺PLGA支架和PLGA膜表面的静态疏水角,发现二者的表面亲水性及湿润性都显著提高了。在此基础上对静电纺PLGA支架进一步进行胶原修饰并得到其扫描电镜图像,发现胶原能够很好的贴附到HFBI修饰后的PLGA支架上。通过在HFBI/胶原共修饰的静电纺PLGA支架上培养细胞,发现细胞在支架上贴附生长状态良好。该结果不仅解决了PLGA材料细胞贴附率低的问题,还为利用PLGA膜制备图案化神经干细胞复合支架器件奠定了基础。因此,利用微接触印刷法将血清印迹到HFBI修饰的PLGA膜上,并在此基底上培养神经干细胞,成功地获得了图案化神经干细胞复合支架。利用HFBI来修饰静电纺PLGA支架和PLGA膜,不仅能有效的改善表/界面材料的亲疏水性,为生物分子、细胞粘附和生长提供便利,还进一步拓展了PLGA在组织工程中的应用。
Biopolymer poly(lacic-co-glycolic acid) PLGA has been extensively applied in tissue engineering as the scaffold materials, due to its good biocompatibility and biodegradability. However, the hydrophobility property of the biopolymer PLGA materials gives rise to the poor cell affinity on the surface and limits their application. Therefore, it is a hot topic to develop an efficient approach to increasing the hydrophilicity of PLGA materials.
In this study, we introduced a class of hydrophobin to modify the biopolymer PLGA. Besides, we also investigated its application both in cell adhesion and the preparation of neural stem cells (NSCs) patterns which made the basis for the fabrication of the complex scaffold composed of NSCs and PLGA film.
Hydrophobins are a group of amphiphilic and surface-active proteins found in filamentous fungi, which has been used to change the surface property of hydrophobic materials. The classⅡhydrophobin HFBI was used to modify electrospun PLGA scaffold and PGLA film. The static water contact angle of the HFBI-modified electrospun PLGA scaffold and PLGA film showed that their surface property conversed from hydrophobictiy to hydrophilicity. Collagen was modified on the HFBI-pretreated electrospun scaffold and the SEM image of the HFBI/collagen co-coated PLGA scaffold demonstrated that collagen could immobilized on the PLGA scaffold with high efficiency. When the HFBI/collagen co-coated PLGA scaffold was used as the substrate of culturing 293 T cells, the cells adhered and grew well on the surface. This result overcame the disadvantage of PLGA materials mentioned above and established the possibility of fabricating neural stem cells patterns on the PLGA film as well. Therefore, the micro-contact printing technology was used to prepare the serum patterns on the HFBI-modified PLGA film and the well-defined neural stem cells patterns was obtained by directly culturing cells on the PLGA film.
HFBI modification of electrospun PLGA scaffold and PLGA film could not only efficiently change the hydrophobicity surface property and facilitate both the immoblization of biomolecular and cell adhesion on the surface, but also enlarge their application in the tissue engineering.
引文
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[3]李双燕,PLGA组织工程支架材料的研究与展望.国外丝绸,2009：29-31
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