Pınarlı Ferda Alpaslan, Okten Gülsen, Beden Umit, Fışgın Tunç, Kefeli Mehmet, Kara Nurten, Duru Feride, Tomak Leman
Department of Medical Biology and Genetics, Faculty of Medicine, Ondokuz Mayis University, Samsun 55270, Turkey.
Department of Ophthalmology, Faculty of Medicine, Ondokuz Mayis University, Samsun 55270, Turkey.
Int J Ophthalmol. 2014 Apr 18;7(2):211-9. doi: 10.3980/j.issn.2222-3959.2014.02.05. eCollection 2014.
To investigate the healing process after severe corneal epithelial damage in rats treated with mesenchymal stem cells (MSCs) cultured with or without keratinocyte growth factor (KGF-2) and autologous serum (AS) on amniotic membrane (AM). Many patients are blind and devastated by severe ocular surface diseases due to limbal stem cell deficiency. Bone marrow-derived MSCs are potential sources for cell-based tissue engineering to repair or replace the corneal tissue, having the potential to differentiate to epithelial cells.
The study included 5 groups each including 10 female "Sprague Dawley" rats in addition to 20 male rats used as bone marrow donors. Group I rats received AM+MSCs, Group II rats AM+MSCs cultured with KGF-2, Group III rats AM+MSCs cultured with KGF-2+AS, Group IV rats only AM and Group V rats, none. AS was derived from blood drawn from male rats and bone marrow was obtained from the femur and tibia bones of the same animals. Therapeutic effect was evaluated with clinical, histopathological and immunohistochemical assessment. MSC engraftment was demonstrated via detection of donor genotype (Y+) in the recipient tissue (X) with polymerase chain reaction.
Corneal healing was significantly better in Groups I-III rats treated with MSC transplantation compared to Group IV and Group V rats with supportive treatment only. The best results were obtained in Group III rats with 90% transparency, 70% lack of neovascularization, and 100% epithelium damage limited to less than 1/4 of cornea.
We suggest that culture of MSCs with KGF-2 and AS on AM is effective in corneal repair in case of irreversible damage to limbal stem cells.
研究在羊膜(AM)上培养的间充质干细胞(MSCs)联合或不联合角质形成细胞生长因子(KGF-2)及自体血清(AS)治疗大鼠严重角膜上皮损伤后的愈合过程。许多患者因角膜缘干细胞缺乏导致严重眼表疾病而失明并深受其害。骨髓来源的间充质干细胞是基于细胞的组织工程修复或替代角膜组织的潜在来源,具有分化为上皮细胞的潜力。
该研究包括5组,每组除20只作为骨髓供体的雄性大鼠外,另有10只雌性“斯普拉格-道利”大鼠。I组大鼠接受AM+MSCs治疗,II组大鼠接受AM+用KGF-2培养的MSCs治疗,III组大鼠接受AM+用KGF-2+AS培养的MSCs治疗,IV组大鼠仅接受AM治疗,V组大鼠不接受任何治疗。AS取自雄性大鼠的血液,骨髓取自同一动物的股骨和胫骨。通过临床、组织病理学和免疫组织化学评估来评价治疗效果。通过聚合酶链反应检测受体组织(X)中的供体基因型(Y+)来证明MSC植入。
与仅接受支持治疗的IV组和V组大鼠相比,接受MSC移植治疗的I-III组大鼠角膜愈合明显更好。III组大鼠取得了最佳结果,角膜透明度达90%,新生血管形成率为70%,上皮损伤局限于角膜不到1/4的区域,占比100%。
我们认为,在角膜缘干细胞发生不可逆损伤的情况下,在羊膜上用KGF-2和AS培养MSCs对角膜修复有效。