Eye Hospital, The First Affiliated Hospital of Harbin Medical University, 23 Youzheng Street, Harbin, 150001, People's Republic of China.
J Mol Histol. 2013 Oct;44(5):609-18. doi: 10.1007/s10735-013-9508-5. Epub 2013 May 1.
The goal of this study was to evaluate the potential suitability of an artificial membrane composed of silk fibroin (SF) functionalized by different ratios of chitosan (CS) as a substrate for the stroma of the cornea. Keratocytes were cultured on translucent membranes made of SF and CS with different ratios. The biophysical properties of the silk fibroin and chitosan (SF/CS) membrane were examined. The SF/CS showed tensile strengths that increased as the CS concentration increased, but the physical and mechanical properties of chitosan-functionalized silk fibroin scaffolds weakened significantly compared with those of native corneas. The resulting cell scaffolds were evaluated using western blot in addition to light and electron microscopy. The cell attachment and proliferation on the scaffold were similar to those on a plastic plate. Keratocytes cultured in serum on SF/CS exhibited stellate morphology along with a marked increase in the expression of keratocan compared with identical cultures on tissue culture plastics. The biocompatibility was tested by transplanting the acellular membrane into rabbit corneal stromal pockets. There was no inflammatory complication detected at any time point on the macroscopic level. Taken together, these results indicate that SF/CS holds promise as a substrate for corneal reconstruction.
本研究旨在评估由不同比例壳聚糖（CS）功能化丝素蛋白（SF）组成的人工膜作为角膜基质的潜在适宜性。将角膜基质细胞培养在具有不同比例 SF 和 CS 的半透明膜上。研究了丝素蛋白和壳聚糖（SF/CS）膜的生物物理特性。SF/CS 的拉伸强度随着 CS 浓度的增加而增加，但与天然角膜相比，壳聚糖功能化丝素蛋白支架的物理和机械性能明显减弱。使用 Western blot 以及光镜和电镜对所得细胞支架进行了评估。支架上的细胞附着和增殖与塑料板上的相似。在 SF/CS 上用血清培养的角膜基质细胞表现出星状形态，并且与在组织培养塑料上的相同培养物相比，角膜蛋白聚糖的表达明显增加。通过将去细胞膜移植到兔角膜基质袋中测试生物相容性。在宏观水平上，任何时间点均未检测到炎症并发症。总之，这些结果表明 SF/CS 有望作为角膜重建的基质。
