Sang Shengbo, Yan Yayun, Shen Zhizhong, Cao Yanyan, Duan Qianqian, He Min, Zhang Qiang
Micro Nano System Research Center, College of Information and Computer & Key Lab of Advanced Transducers and Intelligent Control System of the Ministry of Education, Taiyuan University of Technology, Taiyuan, 030024, China.
Micro Nano System Research Center, College of Information and Computer & Key Lab of Advanced Transducers and Intelligent Control System of the Ministry of Education, Taiyuan University of Technology, Taiyuan, 030024, China; College of Information Science and Engineering, Hebei North University, Zhangjiakou, 075000, China.
Exp Eye Res. 2022 May;218:109027. doi: 10.1016/j.exer.2022.109027. Epub 2022 Mar 8.
The vast majority of patients with corneal blindness cannot recover their vision due to the serious shortage of donor cornea. However, the technology to construct a feasible corneal substitute is a promising treatment method for corneal blindness. In this paper, methacrylated gelatin (GelMA)-methacrylated hyaluronic acid (HAMA) double network (GHDN) hydrogels were prepared by modifying gelatin and hyaluronic acid with methacrylate anhydride (MA). GHDN hydrogel was compared with GelMA single network and HAMA single network hydrogels through characterization experiments of mechanical properties, optical properties, hydrophilicity and in-situ degradation in vitro. At the same time, the biocompatibility of hydrogel was tested by inoculating rabbit corneal epithelial cells (CEpCs) epidermal cells on hydrogels using CCK-8 test, live/dead staining, immunofluorescence staining and qRT-PCR. It was found that the GHDN hydrogel has optical transparency in the visible region, and its mechanical properties are better than those of GelMA and HAMA hydrogels, and its hydrophilicity is similar to that of normal human corneas. The results of in vitro hydrogel culture of CEpCs showed that the proliferation of CEpCs on GHDN hydrogel was two times higher than that of HAMA hydrogel, and the expression of specific marker Cytokeratin 3 (CK3) and Cytokeratin 12 (CK12) could be better maintained on GHDN hydrogel. All the experimental results proved that GHDN hydrogel has good physical properties and biocompatibility and is a potential candidate for corneal tissue engineering scaffolds.
由于供体角膜严重短缺,绝大多数角膜盲患者无法恢复视力。然而,构建可行角膜替代物的技术是治疗角膜盲的一种有前景的方法。本文通过用甲基丙烯酸酐(MA)修饰明胶和透明质酸制备了甲基丙烯酸化明胶(GelMA)-甲基丙烯酸化透明质酸(HAMA)双网络(GHDN)水凝胶。通过力学性能、光学性能、亲水性和体外原位降解的表征实验,将GHDN水凝胶与GelMA单网络和HAMA单网络水凝胶进行了比较。同时,通过CCK-8试验、活/死染色、免疫荧光染色和qRT-PCR在水凝胶上接种兔角膜上皮细胞(CEpCs)表皮细胞来测试水凝胶的生物相容性。结果发现,GHDN水凝胶在可见光区域具有光学透明性,其力学性能优于GelMA和HAMA水凝胶,亲水性与正常人角膜相似。CEpCs水凝胶体外培养结果表明,CEpCs在GHDN水凝胶上的增殖比在HAMA水凝胶上高两倍,且在GHDN水凝胶上能更好地维持特异性标志物细胞角蛋白3(CK3)和细胞角蛋白12(CK12)的表达。所有实验结果证明,GHDN水凝胶具有良好的物理性能和生物相容性,是角膜组织工程支架的潜在候选材料。