a Department of Prosthodontics, Hubei-MOST KLOS & KLOBM , School and Hospital of Stomatology, Wuhan University , Wuhan , China.
b Department of Stomatology, The Central Hospital of Wuhan , Tongji Medical College, Huazhong University of Science and Technology , Wuhan , China.
J Biomater Sci Polym Ed. 2018 Oct;29(15):1859-1875. doi: 10.1080/09205063.2018.1515299. Epub 2018 Oct 7.
Electrospun nanofibers have been considered to be an ideal scaffold for tissue engineering, because of the extracellular-matrix-like structure and the well-controlled fabrication. Here, a new method was used to fabricate electrospun three-dimensional macroporous nanofibrous gelatin scaffolds in ethanol bath by one-step crosslink with glutaraldehyde. The mean diameter of the one-step crosslinked fibers was significantly smaller than that of the traditional two-step crosslinked fibers (p < 0.05), and scaffolds prepared by one-step crosslink were fluffy and porous. No significant difference was found in the degradation rates for both fibers within 14 days. After immersion in PBS for 14 days, numerous two-step crosslinked fibers merged together. By contrast, the morphology and macroporous structure of one-step crosslinked fibers showed no evident change and were generally maintained. Approximate crosslinking degrees of the two-step and one-step crosslinked gelatin fibers were 40% and 54%, respectively (p < 0.05). Results from fluorescence microscopy and hematoxylin-eosin staining showed that MC3T3-E1 subclone four cells were distributed more evenly and diversely in the one-step crosslinked fiber scaffolds. The one-step crosslinked fibers enhanced the proliferation and differentiation potential of MC3T3-E1 cells. Furthermore, one-step crosslinked fibers were beneficial in repairing defects in the skulls of rats. Thus, one-step crosslink by glutaraldehyde in ethanol bath is a cost-effective and simple method to fabricate three-dimensional macroporous nanofiberous scaffolds. This technique retains the morphology and structure of the gelatin fibers, and enhances the biological performance of scaffolds in vitro and in vivo.
静电纺丝纳米纤维因其具有类似于细胞外基质的结构和可精确控制的制备工艺，而被认为是组织工程的理想支架。在这里，采用戊二醛一步交联法在乙醇浴中制备了静电纺丝三维大孔纳米纤维明胶支架。一步交联纤维的平均直径明显小于传统两步交联纤维（p<0.05），且一步交联支架蓬松多孔。在 14 天内，两种纤维的降解速率均无显著差异。在 PBS 中浸泡 14 天后，大量两步交联纤维融合在一起。相比之下，一步交联纤维的形态和大孔结构没有明显变化，基本保持。两步和一步交联明胶纤维的交联度分别约为 40%和 54%（p<0.05）。荧光显微镜和苏木精-伊红染色结果表明，MC3T3-E1 亚克隆 4 细胞在一步交联纤维支架中分布更加均匀和多样。一步交联纤维增强了 MC3T3-E1 细胞的增殖和分化潜能。此外，一步交联纤维有利于修复大鼠颅骨缺陷。因此，戊二醛在乙醇浴中的一步交联是一种经济高效、简单的方法，可以制备三维大孔纳米纤维支架。该技术保留了明胶纤维的形态和结构，增强了支架在体外和体内的生物学性能。
