CAJ | 학술논문

目的：探讨兔骨髓间充质干细胞（ MSCs）移植对兔角膜缘干细胞形态、增殖、分化的影响。方法无菌条件下取兔骨髓，分离得到兔MSCs。取培养3代后MSCs消化后制成细胞悬液，接种于羊膜植片上，常规培养。建立兔角膜碱烧伤干细胞缺损模型4 w后，将18只新西兰兔随机分为空白组、MSCs移植组和模型组，每组6只。MSCs移植组和模型组分别用尼龙缝线将空白和带有兔MSCs的羊膜基质植片间断缝合固定于浅层巩膜上，空白组不做处理。术前、术后观察实验兔眼表形态，并进行评分。术后4 w处死动物，免疫组化检查细胞角蛋白AE5表达情况。结果筛选得到的兔MSCs细胞均一性达98.12%；术后MSCs移植组兔眼表形态综合评分显著降低，而模型组无显著变化，同期与模型组综合评分相比，均显著低于模型组（p<0.05）；免疫组化检测结果显示，空白组和 MSCs 移植组 AE5阳性表达率均为100.00%，显著高于模型组（ p<0.05）。结论以羊膜基质为载体移植MSCs治疗角膜缘干细胞缺乏症，可增殖分化为角膜上皮细胞而发挥功能，疗效显著，可为其临床应用提供理论基础。
목적：탐토토골수간충질간세포（ MSCs）이식대토각막연간세포형태、증식、분화적영향。방법무균조건하취토골수，분리득도토MSCs。취배양3대후MSCs소화후제성세포현액，접충우양막식편상，상규배양。건립토각막감소상간세포결손모형4 w후，장18지신서란토수궤분위공백조、MSCs이식조화모형조，매조6지。MSCs이식조화모형조분별용니룡봉선장공백화대유토MSCs적양막기질식편간단봉합고정우천층공막상，공백조불주처리。술전、술후관찰실험토안표형태，병진행평분。술후4 w처사동물，면역조화검사세포각단백AE5표체정황。결과사선득도적토MSCs세포균일성체98.12%；술후MSCs이식조토안표형태종합평분현저강저，이모형조무현저변화，동기여모형조종합평분상비，균현저저우모형조（p<0.05）；면역조화검측결과현시，공백조화 MSCs 이식조 AE5양성표체솔균위100.00%，현저고우모형조（ p<0.05）。결론이양막기질위재체이식MSCs치료각막연간세포결핍증，가증식분화위각막상피세포이발휘공능，료효현저，가위기림상응용제공이론기출。
Objective To explore the effects of rabbit bone marrow mesenchymal stem cells( MSCs )transplantation on morphology,proliferation,and differentiation of rabbit corneal limbal stem cell. Methods Rabbit bone marrow was obtained under sterile conditions and MSCs were obtained by isolation. Cell suspension was prepared after digestion of MSCs cultured for three generations and inoculated in the amniotic membrane graft,and conventional culture was provided. And then,a rabbit corneal alkali burn stem cell defect model was built. After four weeks,18 New Zealand rabbits were randomly divided into blank group,MSCs transplantation group and model group( n=6 per group). For the MSCs transplantation group and model group,nylon suture was used to fix the amniotic membrane grafts with and without MSCs discontinuously to the superficial sclera,and no treatment was provided for the blank group. The preoperative and postoperative ocular surface morphology of the experimental rabbits were observed and scores were kept. The rabbits were put to death after four weeks since the operation,and immunohistochemistry method was used to examine the expression of cytokeratin AE5. Resuits The homogeneity of rabbit MSCs obtained by screening reached to 98. 12%;The comprehensive scores of postoperative ocular surface morphology of the rabbits in the MSCs transplantation group decreased significantly,while the model group did not change significantly;compared with the model group,the comprehensive scores in the same period were all significantly lower than that in the model group(p <0. 05);immunohistochemistry results showed that the positive expression rates of AE5 in the blank group and MSC transplantation group were 100. 00%,significantly higher than that in the model group(p<0. 05). Conciusion Taking amniotic membrane matrix as a carrier for MSCs transplantation in treatment of corneal limbal stem cell deficiency is available for proliferation and differentiation of corneal epithelial cells,and the curative effect is obvious;this provides theoretical basis for the clinical application.