Yu Jianfeng, Li Pengfei, Fan Kai, Luo Jiawei, Guan Huaijin
Eye Institute, Affiliated Hospital of Nantong University, Nantong, Jiangsu Province, China.
Medical School of Nantong University, Nantong, Jiangsu Province, China.
BMC Ophthalmol. 2025 Jan 20;25(1):30. doi: 10.1186/s12886-025-03873-8.
Human anterior lens capsules (ALCs) have great potential in the treatment of multiple eye diseases, including corneal ulcers, glaucoma, age-related macular degeneration and macular holes. ALCs are also regarded as promising scaffolds for various ocular cells. Here, we investigated different decellularization methods for removing lens epithelial cells (LECs) that adhered to ALCs.
Human ALCs were treated with various solutions, including 2% lidocaine, 10% sodium chloride, 50% glucose and sterile water. Trypan blue and alizarin red (TB-AR) staining, H&E staining and hydroxyproline assays were used to assess the degree of decellularization. The impacts of acellular ALCs on cell viability and cell-tissue interaction were investigated in vitro.
These findings revealed that 2% lidocaine, 10% sodium chloride, 50% glucose, and sterile water had the capacity to decellularize ALCs at 37 °C. The structure of ALCs in all decellularization groups was similar to that of intact ALCs. The effects of 10% sodium chloride and sterile water on decellularization were significantly better than those of 2% lidocaine and 50% glucose. The H&E staining revealed that the different decellularization methods maintained the integrity of the lens capsular structure. Compared with sterile water, 10% sodium chloride preserved a better level of hydroxyproline. The ALCs in the 10% sodium chloride-treated group and the sterile water-treated group were shown to be suitable for cell adhesion in vitro.
This study identified two optimal decellularization methods for acellular ALCs: using 10% sodium chloride and using sterile water. The obtained acellular ALCs could be promising scaffolds for ocular cells. In addition, acellular ALCs do not need resterilization and may be directly used for autologous lens capsule transplantation in clinical applications.
人眼前囊膜在多种眼科疾病的治疗中具有巨大潜力,包括角膜溃疡、青光眼、年龄相关性黄斑变性和黄斑裂孔。人眼前囊膜也被视为各种眼细胞有前景的支架材料。在此,我们研究了不同的去细胞方法以去除附着于人眼前囊膜上的晶状体上皮细胞(LECs)。
用人眼前囊膜分别用多种溶液处理,包括2%利多卡因、10%氯化钠、50%葡萄糖和无菌水。采用台盼蓝和茜素红(TB-AR)染色、苏木精-伊红(H&E)染色和羟脯氨酸测定来评估去细胞程度。在体外研究了脱细胞人眼前囊膜对细胞活力和细胞-组织相互作用的影响。
这些发现表明,2%利多卡因、10%氯化钠、50%葡萄糖和无菌水在37℃时均有能力使前囊膜去细胞。所有去细胞组的前囊膜结构与完整前囊膜相似。10%氯化钠和无菌水的去细胞效果明显优于2%利多卡因和50%葡萄糖。H&E染色显示,不同的去细胞方法保持了晶状体囊膜结构的完整性。与无菌水相比,10%氯化钠保留了更好水平的羟脯氨酸。10%氯化钠处理组和无菌水处理组的前囊膜在体外显示适合细胞黏附。
本研究确定了两种用于脱细胞人眼前囊膜的最佳去细胞方法:使用10%氯化钠和使用无菌水。所获得的脱细胞人眼前囊膜可能是有前景的眼细胞支架材料。此外,脱细胞人眼前囊膜无需再次灭菌,在临床应用中可直接用于自体晶状体囊膜移植。