Effects of UVA+UVB Solar Radiation Levels and Cross-Linking on Animal Model with Bowman’s Layer: Chicken

Marîa Carmen Martínez García
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Abstract

Effects of UVA+ UVB Solar Radiation Levels and Cross-Linking on Animal Model with Bowman’s Layer: Chicken. Clin Res 6: 059. Abstract Purpose: Evaluate the corneal damage and wound healing in an animal model with Bowman’s layer after UVA+UVB radiation and epi-off Cross-Linking (CXL). Methods: Adult Brown Classic chickens were randomly divided into three groups. The UVA+UVB group (20 eyes) was exposed to three doses of radiation (4.60 J/cm 2 ). The CXL group (12 eyes), was de-epithelialized and irradiated with UVA light (5.4 J/cm 2 ) for 30 minutes. The contralateral eyes were used as a control (12 eyes). The animals have clinically follow-up, the epithelial closure was measured by fluorescein stain and corneal thickness by pachymetry. Keratocytes loss, subsequent cell repopulation, and healing events were analyzed after euthanasia at 24 hours, 15 and 30 days post-ra- diation on hematoxylin-eosin-stained sections. Cell death was detected by TUNEL assay, proliferation by BrdU (bromodeoxyuridine) immunofluorescence, and myofibroblasts differentiation by (αSMA) immunohistochemistry. Results: Re-epithelialization was at 5 ± 2.3 days in CXL and at day 2 ± 0.9 days in UVA+UVB. Pachymetry was higher after UVA+UVB than CXL (p<0.05). Cell death affected the whole thickness of stroma in the UVA+UVB group and anterior and middle stroma in CXL. At day 1, the UVA+UVB group showed a more significant reduction in the number of stromal cells. After 30 days, both groups showed a free band of cells below the epithelium. Conclusion: The damage and the healing response in chicken cor- neas after UVA+UVB and CXL demonstrated that Bowman’s layer has not been successful in protecting the cornea from radiation. Perhaps, other factors that are involved in corneal defense.
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UVA+UVB太阳辐射水平及交联对鲍曼综合征鸡模型的影响
UVA+ UVB太阳辐射水平及交联对鲍曼综合征鸡模型的影响临床研究6:059。目的:评价UVA+UVB辐射和epi-off交联(CXL)对鲍曼层动物模型角膜损伤和创面愈合的影响。方法:将成年布朗经典鸡随机分为3组。UVA+UVB组(20只眼睛)暴露于三次剂量的辐射(4.60 J/ cm2)。CXL组(12只眼)去上皮,UVA光(5.4 J/ cm2)照射30分钟。对侧眼作为对照(12只眼)。对实验动物进行临床随访,荧光素染色测定上皮闭合程度,角膜厚测法测定角膜厚度。在苏木精-伊红染色切片上分析安乐死后24小时、15天和30天的角质细胞损失、随后的细胞再生和愈合事件。TUNEL法检测细胞死亡,BrdU(溴脱氧尿苷)免疫荧光法检测细胞增殖,(αSMA)免疫组化法检测肌成纤维细胞分化。结果:CXL的再上皮化时间为5±2.3天,UVA+UVB的再生时间为2±0.9天。UVA+UVB治疗后血肿明显高于CXL (p<0.05)。细胞死亡影响了UVA+UVB组间质整体厚度和CXL前中层间质厚度。在第1天,UVA+UVB组间质细胞数量减少更为显著。30 d后,两组上皮下均出现游离带。结论:UVA+UVB和CXL对鸡角膜的损伤和愈合反应表明,鲍曼层对角膜的保护作用并不成功。也许还有其他因素与角膜防御有关。
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