中华眼科杂志
中華眼科雜誌
중화안과잡지
Chinese Journal of Ophthalmology
2014年
4期
277-284
,共8页
朱勤%胡竹林%孙晓梅%胡敏%刘海%杨忠昆
硃勤%鬍竹林%孫曉梅%鬍敏%劉海%楊忠昆
주근%호죽림%손효매%호민%류해%양충곤
内皮,血管%内皮,角膜%内皮细胞%细胞移植%显微镜检查,电子,扫描
內皮,血管%內皮,角膜%內皮細胞%細胞移植%顯微鏡檢查,電子,掃描
내피,혈관%내피,각막%내피세포%세포이식%현미경검사,전자,소묘
Endothelium,vascular%Endothelium,corneal%Endothelial cells%Cell transplantation%Microscopy,electron,scanning
目的 探讨血管内皮细胞(VEC)移植治疗角膜内皮损伤的可行性.方法 实验研究.体外培养增殖VEC.将恒河猴12只,按照随机数字表随机分为两组:实验组(6只)、对照组(6只).实验组通过超声乳化破坏角膜内皮细胞层之后,将体外培养的VEC利用前房注射法移植到恒河猴角膜内表面;对照组超声乳化破坏角膜内皮细胞层后不注射VEC.术后7、14、30、60、90 d分别行术眼前节照相;术前及术后7、14、30、60、90 d分别用A超测量角膜厚度、Goldman眼压计测量眼压,对数据进行独立样本t检验及方差分析;术后30、60、90 d分别摘取术眼行病理切片HE染色、扫描电镜观察VEC在角膜植片内表面的分布.结果 实验组角膜比对照组有较好的透明度,未出现大泡性角膜病变,术后3个月时实验组角膜缘可见少量新生血管.术眼前房深度均正常;术后3个月时,实验组角膜厚度为(500±14) μm,明显低于对照组角膜厚度[(618±11)μm],同时期实验组和对照之间角膜植片厚度值变化差异有统计学意义(术后1、2、3个月t值分别为-3.256、-4.419、-12.896,均P＜0.05);同时期实验组和对照之间眼压变化差异无统计学意义(术后1、2、3个月t值分别为-1.179、-2.166、-2.536,均P＞0.05);实验组角膜内表面可见细胞层;对照组角膜内表面未见细胞样结构.扫描电镜:实验组角膜植片内表面可见VEC分布均匀,生长良好,呈不规则多边形,表面可见微绒毛,VEC间可见少量白细胞,小梁网处部分细胞碎片聚集;对照组角膜内表面呈纤维物质样,未见细胞生长.结论 超声乳化法能很好的建立恒河猴角膜内皮损伤模型;前房注射法能够将VEC移植到角膜内表面,且细胞能在撕出后弹力层的角膜内表面生长,并在一定程度上发挥屏障作用维持角膜的脱水状态和透明性.移植手术对前房角的组织结构形态在短期内不产生病理性影响.
目的 探討血管內皮細胞(VEC)移植治療角膜內皮損傷的可行性.方法 實驗研究.體外培養增殖VEC.將恆河猴12隻,按照隨機數字錶隨機分為兩組:實驗組(6隻)、對照組(6隻).實驗組通過超聲乳化破壞角膜內皮細胞層之後,將體外培養的VEC利用前房註射法移植到恆河猴角膜內錶麵;對照組超聲乳化破壞角膜內皮細胞層後不註射VEC.術後7、14、30、60、90 d分彆行術眼前節照相;術前及術後7、14、30、60、90 d分彆用A超測量角膜厚度、Goldman眼壓計測量眼壓,對數據進行獨立樣本t檢驗及方差分析;術後30、60、90 d分彆摘取術眼行病理切片HE染色、掃描電鏡觀察VEC在角膜植片內錶麵的分佈.結果 實驗組角膜比對照組有較好的透明度,未齣現大泡性角膜病變,術後3箇月時實驗組角膜緣可見少量新生血管.術眼前房深度均正常;術後3箇月時,實驗組角膜厚度為(500±14) μm,明顯低于對照組角膜厚度[(618±11)μm],同時期實驗組和對照之間角膜植片厚度值變化差異有統計學意義(術後1、2、3箇月t值分彆為-3.256、-4.419、-12.896,均P＜0.05);同時期實驗組和對照之間眼壓變化差異無統計學意義(術後1、2、3箇月t值分彆為-1.179、-2.166、-2.536,均P＞0.05);實驗組角膜內錶麵可見細胞層;對照組角膜內錶麵未見細胞樣結構.掃描電鏡:實驗組角膜植片內錶麵可見VEC分佈均勻,生長良好,呈不規則多邊形,錶麵可見微絨毛,VEC間可見少量白細胞,小樑網處部分細胞碎片聚集;對照組角膜內錶麵呈纖維物質樣,未見細胞生長.結論 超聲乳化法能很好的建立恆河猴角膜內皮損傷模型;前房註射法能夠將VEC移植到角膜內錶麵,且細胞能在撕齣後彈力層的角膜內錶麵生長,併在一定程度上髮揮屏障作用維持角膜的脫水狀態和透明性.移植手術對前房角的組織結構形態在短期內不產生病理性影響.
목적 탐토혈관내피세포(VEC)이식치료각막내피손상적가행성.방법 실험연구.체외배양증식VEC.장항하후12지,안조수궤수자표수궤분위량조:실험조(6지)、대조조(6지).실험조통과초성유화파배각막내피세포층지후,장체외배양적VEC이용전방주사법이식도항하후각막내표면;대조조초성유화파배각막내피세포층후불주사VEC.술후7、14、30、60、90 d분별행술안전절조상;술전급술후7、14、30、60、90 d분별용A초측량각막후도、Goldman안압계측량안압,대수거진행독립양본t검험급방차분석;술후30、60、90 d분별적취술안행병리절편HE염색、소묘전경관찰VEC재각막식편내표면적분포.결과 실험조각막비대조조유교호적투명도,미출현대포성각막병변,술후3개월시실험조각막연가견소량신생혈관.술안전방심도균정상;술후3개월시,실험조각막후도위(500±14) μm,명현저우대조조각막후도[(618±11)μm],동시기실험조화대조지간각막식편후도치변화차이유통계학의의(술후1、2、3개월t치분별위-3.256、-4.419、-12.896,균P＜0.05);동시기실험조화대조지간안압변화차이무통계학의의(술후1、2、3개월t치분별위-1.179、-2.166、-2.536,균P＞0.05);실험조각막내표면가견세포층;대조조각막내표면미견세포양결구.소묘전경:실험조각막식편내표면가견VEC분포균균,생장량호,정불규칙다변형,표면가견미융모,VEC간가견소량백세포,소량망처부분세포쇄편취집;대조조각막내표면정섬유물질양,미견세포생장.결론 초성유화법능흔호적건립항하후각막내피손상모형;전방주사법능구장VEC이식도각막내표면,차세포능재시출후탄력층적각막내표면생장,병재일정정도상발휘병장작용유지각막적탈수상태화투명성.이식수술대전방각적조직결구형태재단기내불산생병이성영향.
Objective To observe form and function changes of vascular endothelial cells (VEC) which were transplanted to the posterior surface of rhesus monkey cornea without Descemet's Membrane by anterior chamber injection,explore the feasibility of transplanting VEC to treat corneal endothelial injury,and find new method of corneal endothelial cell transplantation.Methods Cultured VEC to proliferate in vitro.Rhesus monkeys were randomly divided into two groups:Experimental group(6) and control group(6)according to a random number table.The experimental group:transplant the culured VEC to the posterior surface of rhesus monkey cornea without Descemet's membrane by anterior chamber injection.The control group:Tear out the Descemet's membrane by capsulotomy needle without VEC transplantation.A ultrasound apparatus was adopted to measure the postoperative thicknes of the cornea and Goldman intraocular pressure meter measuring intraocular pressure in the postoperative 7 days,14 days,30 days,60 days,90 days,organize the data and entered into the computer,applicated the software of SPSS 11.5 for data independent samples t-test and analysis of variance.The eyes were removed respectively in postoperative 30,60,90 days to do pathological HE dyeing and scanning electron microscopy (SEM) observation of VEC in the posterior surface of cornea graft.Results Corneal transpareney:In the experimental observation period (3 months),the experimental group had better transparency than the control group with normal anterior chamber depth and without bullous kerat-opathy.And the corneal neovascularization was exist in the cornea graft in experimental group in the third month.After 3 months,the corneal thickness of experimental group (500 ± 14) μm was significantly lower than the control group (618 ± 11) μm,Corneal thickness values between experimental and control groups were statistically significant differences in changes (all P ＜ 0.05,t-values were-3.256,-4.419,-12.896 postoperative 1,2,3 months).Changes of intraocular pressure difference between the experimental group and the control was not statistically significant (all P ＞ 0.05,t-values were-1.179,-2.166,-2.536 postoperative 1,2,3 months).The pathological:the cell layer was visible in the posterior surface of cornea graft.The control group:can't find the cell sample structure.SEM:Experimental group showed that VEC with irregular shape uniformly distributed on the inner surface of cornea and growing well,a small amount of white blood cells can be seen between VEC,and part of cellular debris exist in the trabecular meshwork.Control group showed a fiber material without VEC.Conclusions Ultrasonic emulsification can established a repeatability and simple model of corneal endothelial injury in Rhesus monkeys.VEC can be transplanted to the corneal surface by Anterior chamber injection and the cells can grow on the surface and play a barrier role in maintaining the state of dehydration and transparency of the cornea to a certain extent.After transplantation,organizational structure and morphology of the anterior chamber angle does not produce pathological effects in the short term.Prompt that transplant the culured VEC to the posterior surface of rhesus monkey cornea without Descemet's Membrane by anterior chamber injection to substitute the function of the corneal endothelial cells may be a new idea for treatment of cornealendothelial damage.
