Yan Chao, Wang Li, Pan Zhi-qiang
Beijing Tongren Eye Bank, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital University of Medical Sciences, Beijing 100730, China.
Zhonghua Yan Ke Za Zhi. 2006 Feb;42(2):166-70.
To investigate the effect of basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) on corneal endothelial cell preserved in mid-term preserving medium.
Corneal rings were preserved in mid-term preserving medium in control group. bFGF (5 ng/ml), bFGF (20 ng/ml), EGF and bFGF + EGF were added into the mid-term preserving medium in experimental groups A, B, C, D respectively. At the 3rd, 7th and 14th day after preservation, the viability of corneal endothelial cells (CECs) was assessed by trypan blue and alizarin red stain. The microstructure of corneal endothelial cells was observed by electron microscope (EM).
At the 14th days of preservation, viability of CECs was (10.35 +/- 1.32)% (control group), (62.18 +/- 1.56)% (group A), (92.57 +/- 0.90)% (group B), (71.01 +/- 2.67)% (group C) and (82.59 +/- 1.45)% in group D respectively. The viability of CECs in experimental groups was much higher than in control group (P < 0.01). At the 14th days, corneal rings in control group showed edematous, cloud, defect of integrity of endothelial cells and Y type conjunction broken. But in experimental groups, the corneal edema and folds were slightly, furthermore the cornea ring was transparency and cell shape was intact, especially in group B and group D. Electronic microscope (EM) showed that corneal endothelial cells Y type conjunction in experimental groups was completely integrity, endothelial cells had tight conjunction and rich micro-villa on surface, large cells body and obvious nucleus.
Addition of bFGF and EGF in corneal storage medium plays a significant role in maintaining CECs structure and viability.
探讨碱性成纤维细胞生长因子(bFGF)和表皮生长因子(EGF)对中期保存液中保存的角膜内皮细胞的作用。
对照组角膜环在中期保存液中保存。实验组A、B、C、D分别在中期保存液中加入bFGF(5 ng/ml)、bFGF(20 ng/ml)、EGF以及bFGF + EGF。保存后第3天、第7天和第14天,通过台盼蓝和茜素红染色评估角膜内皮细胞(CECs)的活性。通过电子显微镜(EM)观察角膜内皮细胞的微观结构。
保存第14天时,CECs活性在对照组为(10.35±1.32)%,实验组A为(62.18±1.56)%,实验组B为(92.57±0.90)%,实验组C为(71.01±2.67)%,实验组D为(82.59±1.45)%。实验组CECs活性显著高于对照组(P < 0.01)。第14天时,对照组角膜环出现水肿、浑浊、内皮细胞完整性缺陷以及Y型连接断裂。但实验组角膜水肿和褶皱较轻,角膜环透明且细胞形态完整,尤其是实验组B和实验组D。电子显微镜(EM)显示实验组角膜内皮细胞Y型连接完全完整,内皮细胞连接紧密,表面有丰富的微绒毛,细胞体大且细胞核明显。
在角膜保存液中添加bFGF和EGF对维持CECs结构和活性具有重要作用。