Zeppieri Marco, Salvetat Maria Letizia, Beltrami Antonio, Cesselli Daniela, Russo Rossella, Alcalde Ignacio, Merayo-Lloves Jesús, Brusini Paolo, Parodi Pier Camillo
Department of Ophthalmology, Azienda Ospedaliero Universitaria Santa Maria della Misericordia, Udine 33100, Italy.
Department of Pathology, University of Udine, Azienda Ospedaliero Universitaria Santa Maria della Misericordia, Udine 33100, Italy.
J Clin Med. 2017 Dec 5;6(12):115. doi: 10.3390/jcm6120115.
The aim of our study was to assess the clinical effectiveness of topical adipose derived stem cell (ADSC) treatment in laser induced corneal wounds in mice by comparing epithelial repair, inflammation, and histological analysis between treatment arms. Corneal lesions were performed on both eyes of 40 mice by laser induced photorefractive keratectomy. All eyes were treated with topical azythromycin bid for three days. Mice were divided in three treatment groups ( = 20), which included: control, stem cells and basic serum; which received topical treatment three times daily for five consecutive days. Biomicroscope assessments and digital imaging were performed by two masked graders at 30, 54, 78, 100, and 172 h to analyze extent of fluorescein positive epithelial defect, corneal inflammation, etc. Immunohistochemical techniques were used in fixed eyes to assess corneal repair markers Ki67, α Smooth Muscle Actin (α-SMA) and E-Cadherin. The fluorescein positive corneal lesion areas were significantly smaller in the stem cells group on days 1 ( < 0.05), 2 ( < 0.02) and 3. The stem cell treated group had slightly better and faster re-epithelization than the serum treated group in the initial phases. Comparative histological data showed signs of earlier and better corneal repair in epithelium and stromal layers in stem cell treated eyes, which showed more epithelial layers and enhanced wound healing performance of Ki67, E-Cadherin, and α-SMA. Our study shows the potential clinical and histological advantages in the topical ADSC treatment for corneal lesions in mice.
我们研究的目的是通过比较各治疗组之间的上皮修复、炎症反应和组织学分析,评估局部应用脂肪来源干细胞(ADSC)治疗小鼠激光诱导角膜伤口的临床效果。通过激光诱导的准分子激光原位角膜磨镶术在40只小鼠的双眼上造成角膜损伤。所有眼睛均局部应用阿奇霉素,每日两次,持续三天。将小鼠分为三个治疗组(每组n = 20),包括:对照组、干细胞组和基础血清组;连续五天每天进行三次局部治疗。由两名不知情的评分者在30、54、78、100和172小时进行生物显微镜评估和数字成像,以分析荧光素阳性上皮缺损的程度、角膜炎症等。对固定后的眼睛采用免疫组织化学技术评估角膜修复标志物Ki67、α平滑肌肌动蛋白(α-SMA)和E-钙黏蛋白。在第1天(P < 0.05)、第2天(P < 0.02)和第3天,干细胞组的荧光素阳性角膜损伤面积明显较小。在初始阶段,干细胞治疗组的再上皮化比血清治疗组略好且更快。比较组织学数据显示,干细胞治疗的眼睛在角膜上皮和基质层有更早且更好的角膜修复迹象,表现为上皮层数更多,Ki67、E-钙黏蛋白和α-SMA的伤口愈合性能增强。我们的研究显示了局部应用ADSC治疗小鼠角膜损伤在临床和组织学方面的潜在优势。