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用于角膜上皮重建的无载体培养自体口腔黏膜上皮细胞片(CAOMECS):组织学研究。

Carrier-free cultured autologous oral mucosa epithelial cell sheet (CAOMECS) for corneal epithelium reconstruction: a histological study.

机构信息

Department of Medicine, LA BioMed at Harbor UCLA Medical Center, Torrance, CA, USA.

Department of Medicine, LA BioMed at Harbor UCLA Medical Center, Torrance, CA, USA.

出版信息

Ocul Surf. 2015 Apr;13(2):150-63. doi: 10.1016/j.jtos.2014.12.003. Epub 2015 Jan 30.

Abstract

This study investigates the therapeutic effects of carrier-free cultured autologous oral mucosa epithelial cell sheet (CAOMECS) transplantation for experimentally induced severe rabbit limbal stem cell deficiency (LSCD). Buccal biopsies were performed and CAOMECS were cultured and transplanted onto diseased corneas. Six-month follow-up examinations indicated that three out of four corneas with CAOMECS grafts showed a decrease in superficial vascularization, while almost all the sham corneas did not show a similar decrease. H&E staining of corneas showed that CAOMECS transplantation reduced blood vessel invasion of central cornea, reduced lymphocyte infiltration and fibrotic tissue formation. DeltaNp63 stained markedly in the grafted cornea and to a lesser extent in the sham corneas. PCNA and Ki-67 staining were much greater in the sham corneas than in the grafted and normal corneas. K3 and K13 staining demonstrated that CAOMECS transplanted corneas had much more K3- and less K13- positive cells compared to the sham corneas. Muc5AC was decreased in the central region of grafted corneas. Very little alpha-smooth muscle actin (aSMA) staining was detected in grafted corneas, while there was a greater amount of aSMA staining in sham corneas. Staining for anti-angiogenic factor TIMP -3 was also increased, and pro-angiogenic factor MMP-3 was decreased in grafted corneas compared to sham corneas. Our results indicate that CAOMECS grafts resulted in improved epithelialization of the corneal surface and decreased vascularization and fibrosis of the diseased corneas.

摘要

本研究探讨了无载体培养的自体口腔黏膜上皮细胞片(CAOMECS)移植治疗实验性诱导的严重兔角膜缘干细胞缺乏症(LSCD)的治疗效果。进行颊部活检,并培养和移植 CAOMECS 到病变角膜上。6 个月的随访检查表明,四分之三的 CAOMECS 移植物角膜的浅层血管化减少,而几乎所有的假手术角膜均未显示出类似的减少。角膜的 H&E 染色显示,CAOMECS 移植减少了中央角膜的血管侵犯,减少了淋巴细胞浸润和纤维组织形成。DeltaNp63 在移植物角膜中染色明显,在假手术角膜中染色程度较低。PCNA 和 Ki-67 染色在假手术角膜中比在移植物和正常角膜中更大。K3 和 K13 染色表明,与假手术角膜相比,CAOMECS 移植角膜的 K3 阳性细胞更多,K13 阳性细胞更少。Muc5AC 在移植物角膜的中央区域减少。移植物角膜中检测到的α-平滑肌肌动蛋白(aSMA)染色很少,而假手术角膜中 aSMA 染色更多。抗血管生成因子 TIMP-3 的染色也增加,与假手术角膜相比,移植物角膜中促血管生成因子 MMP-3 减少。我们的结果表明,CAOMECS 移植物导致角膜表面上皮化改善,病变角膜的血管化和纤维化减少。

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