Wan Lu-Qin, Zhang Yang-Yang, Wang Hui-Feng, Chen Chen, Li Hua, Zhang Yuan, Xue Jun-Fa, Zhou Qing-Jun, Xie Li-Xin
Medical College of Qingdao University, Qingdao 266000, Shandong Province, China.
State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao 266071, Shandong Province, China.
Int J Ophthalmol. 2021 Nov 18;14(11):1653-1659. doi: 10.18240/ijo.2021.11.02. eCollection 2021.
To evaluate the efficacy of recombinant human nerve growth factor-loaded amniotic membrane (rhNGF-AM) on corneal epithelial and nerve regeneration in rabbit model.
Freshly prepared human amniotic membrane (AM) were immersed into PBS buffer containing 100 or 500 µg/mL rhNGF for 15, 30, and 60min at 4°C. The release kinetics of rhNGF was measured with ELISA. For evaluation, the AM were immersed with 500 µg/mL rhNGF for 30min. Fifty-seven rabbits were selected to establish corneal epithelial defect model. In addition to the 19 rabbits in control group, 38 rabbits received AM transplantation with or without rhNGF after the removal of central epithelium. Corneal epithelial defect area, sub-epithelial nerve fiber density, corneal sensitivity, rhNGF contents in resident AM and corneas were measured after the surgery.
rhNGF was sustained release from the AM within 14d , with the positive correlation with initial immersion concentration. The immersion of AM in 500 µg/mL rhNGF for 30min achieved the most stable release within 14d. After transplantation in rabbit cornea, a high concentration of rhNGF in resident rhNGF-AM and cornea was maintained within 8d. Corneal epithelial healing, nerve fiber regeneration and the recovery of corneal sensitivity were significantly accelerated after the rhNGF-AM transplantation when compared to simple AM transplantation (all <0.05).
Simple immersion of AM achieves the sustained release of rhNGF, and promotes corneal epithelial wound healing and nerve regeneration, as well as the recovery of corneal sensitivity in rabbit.
评估负载重组人神经生长因子的羊膜（rhNGF-AM）对兔角膜上皮及神经再生的疗效。
将新鲜制备的人羊膜（AM）于4℃下分别浸入含100或500μg/mL rhNGF的PBS缓冲液中15、30和60分钟。采用ELISA法测定rhNGF的释放动力学。为进行评估，将羊膜浸入500μg/mL rhNGF中30分钟。选取57只兔建立角膜上皮缺损模型。除对照组的19只兔外，38只兔在去除中央上皮后接受含或不含rhNGF的羊膜移植。术后测量角膜上皮缺损面积、上皮下神经纤维密度、角膜敏感性、驻留羊膜和角膜中的rhNGF含量。
rhNGF在14天内从羊膜中持续释放，与初始浸泡浓度呈正相关。将羊膜浸入500μg/mL rhNGF中30分钟在14天内实现了最稳定的释放。在兔角膜移植后，驻留rhNGF-AM和角膜中的高浓度rhNGF在8天内得以维持。与单纯羊膜移植相比，rhNGF-AM移植后角膜上皮愈合、神经纤维再生及角膜敏感性恢复均显著加快（均P<0.05）。
羊膜简单浸泡可实现rhNGF的持续释放，并促进兔角膜上皮伤口愈合、神经再生及角膜敏感性恢复。
