家兔角膜缘干细胞缺失病理模型的制作

Preparation of pathological model of rabbit total corneal limbal stem cell deficiency

根据角膜缘干细胞定位于角膜缘上皮基底层,为角膜上皮增殖分化的源泉并保持角膜上皮完整性的理论。将20只家兔随机分为4组,每组5只,分别将角膜缘上皮全周或半周板层手术切除,或用1mol/LNaOH直接擦除等方法破坏角膜缘干细胞,探索制作角膜缘干细胞完全缺失病理模型的有效途径。结果表明,处理后4周,全周角膜缘上皮板层手术切除,中央角膜上皮层用1mol/LNaOH擦除的5只试验家兔角膜表面全部血管化、结膜化,未发生睑球粘连,角膜基质胶原纤维完整未见溃疡、穿孔等病变,细胞印迹学检查为结膜表型,可作为实验性角膜缘干细胞移植的病理模型;全周角膜缘上皮板层手术切除,中央角膜上皮用生理盐水擦除的5只试验家兔,有2只为结膜表型,另3只为角膜表型,观察期内结果不稳定;半周角膜缘上皮板层手术切除,中央角膜上皮层用生理盐水擦除的5只试验家兔,角膜表面透明,全部为角膜表型;直接用1mol/LNaOH擦除角膜缘和中央角膜上皮的试验家兔,有4只角膜基质胶原纤维断裂、溶解,并伴有严重的溃疡、穿孔、睑球粘连等病变,不能用于移植试验,另1只角膜表面透明,未见结膜和新生血管长入,细胞印迹学检查为角膜表型。

According to the theory that corneal epithelial stem cell locates at limbus and serves as the ultimate source for maintenance and regeneration of the corneal epithelium,20 experimental rabbits were randomly divided into four groups,5 rabbits in each group.The whole or upper-half of corneal limbal epithelium lamella were excised by surgery or burned with 1 mol/L NaOH for exploring the best creating approach of the pathological model of corneal total limbal stem cell deficiency.The results showed,4 weeks after operation,that the methods of the corneal limbal epithelium lamella were excised completely,and the center corneal epitheliums burned with 1 mol/L NaOH could successfully create the physiological model of limbal stem cell total deficiency;all 5 rabbits corneal surface were haze with neovascularization and conjunctivalizaion,the corneal stroma collagen were integrant,symblepharon (adhesions of lid conjunctiva to globe conjunctiva) were not observed,which could be used for limbal stem cells transplants;At the same time,the whole corneal limbal epithelium lamella were excised and the center corneal epitheliums were erased with a swab soaked in physiological saline,there were 2 rabbits corneal surface presented corneal epithelium phenotype,and the other 3 presented conjunctiva epithelium phenotype.The results are unsure for experimental use;The rabbit's corneas that were removed with upper-half of corneal limbal epithelium lamella and erased the center corneal epitheliums were transparent with intact corneal epithelium;In the approach,the corneal and limbal epitheliums were burned with a cotton swab socked in 1 mol/L NaOH,there were 4 rabbits' corneal stroma happened perforation or ulcer and symblepharon,and the other one presented corneal epithelium phenotype.This is an applicable method to create the pathological model of corneal limbal stem cell total deficiency.