Stafiej Piotr, Küng Florian, Thieme Daniel, Czugala Marta, Kruse Friedrich E, Schubert Dirk W, Fuchsluger Thomas A
Department of Ophthalmology, Universität Erlangen-Nürnberg, Schwabachanlage 6, 91054 Erlangen, Germany; Institute of Polymer Materials, Universität Erlangen-Nürnberg, Martensstraße 7, 91054 Erlangen, Germany.
Department of Ophthalmology, Universität Erlangen-Nürnberg, Schwabachanlage 6, 91054 Erlangen, Germany.
Mater Sci Eng C Mater Biol Appl. 2017 Feb 1;71:764-770. doi: 10.1016/j.msec.2016.10.058. Epub 2016 Oct 26.
In this work, polycaprolactone (PCL) was used as a basic polymer for electrospinning of random and aligned nanofiber matrices. Our aim was to develop a biocompatible substrate for ophthalmological application to improve wound closure in defects of the cornea as replacement for human amniotic membrane. We investigated whether blending the hydrophobic PCL with poly (glycerol sebacate) (PGS) or chitosan (CHI) improves the biocompatibility of the matrices for cell expansion. Human corneal epithelial cells (HCEp) and human corneal keratocytes (HCK) were used for in vitro biocompatibility studies. After optimization of the electrospinning parameters for all blends, scanning electron microscopy (SEM), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), and water contact angle were used to characterize the different matrices. Fluorescence staining of the F-actin cytoskeleton of the cells was performed to analyze the adherence of the cells to the different matrices. Metabolic activity of the cells was measured by cell counting kit-8 (CCK-8) for 20days to compare the biocompatibility of the materials. Our results show the feasibility of producing uniform nanofiber matrices with and without orientation for the used blends. All materials support adherence and proliferation of human corneal cell lines with oriented growth on aligned matrices. Although hydrophobicity of the materials was lowered by blending PCL, no increase in biocompatibility or proliferation, as was expected, could be measured. All tested matrices supported the expansion of human corneal cells, confirming their potential as substrates for biomedical applications.
在本研究中，聚己内酯（PCL）被用作静电纺丝制备无规和取向纳米纤维基质的基础聚合物。我们的目标是开发一种用于眼科应用的生物相容性基质，以改善角膜缺损处的伤口闭合，作为人羊膜的替代品。我们研究了将疏水性PCL与聚（癸二酸甘油酯）（PGS）或壳聚糖（CHI）共混是否能提高基质对细胞扩增的生物相容性。人角膜上皮细胞（HCEp）和人角膜基质细胞（HCK）用于体外生物相容性研究。在优化所有共混物的静电纺丝参数后，使用扫描电子显微镜（SEM）、衰减全反射傅里叶变换红外光谱（ATR-FTIR）和水接触角来表征不同的基质。对细胞的F-肌动蛋白细胞骨架进行荧光染色，以分析细胞对不同基质的粘附情况。使用细胞计数试剂盒-8（CCK-8）测量细胞的代谢活性，持续20天，以比较材料的生物相容性。我们的结果表明，对于所使用的共混物，制备有取向和无取向的均匀纳米纤维基质是可行的。所有材料都支持人角膜细胞系的粘附和增殖，且在取向基质上呈取向生长。尽管通过与PCL共混降低了材料的疏水性，但并未如预期那样检测到生物相容性或增殖的增加。所有测试的基质都支持人角膜细胞的扩增，证实了它们作为生物医学应用基质的潜力。
