Anitua Eduardo, Muruzabal Francisco, Pino Ander, Merayo-Lloves Jesús, Orive Gorka
*Fundación Eduardo Anitua, Vitoria, Spain; and †Fundación de Investigación Oftalmológica, Instituto Oftalmológico Fernández-Vega, Oviedo, Spain.
Cornea. 2013 Oct;32(10):1380-6. doi: 10.1097/ICO.0b013e31829f7088.
We evaluated whether plasma rich in growth factors eye drops maintain their composition and biological activity after storage for 3 months at -20°C and after storage at 4°C or room temperature (RT) for 24 hours, compared with samples obtained at time 0 (fresh samples).
Blood from 10 healthy donors was collected, centrifuged, and plasma rich in growth factors was prepared by avoiding the collection of the buffy coat. Eye drops were kept fresh or were stored at -20°C for 15, 30, and 90 days. For each time, 2 aliquots were kept at RT or at 4°C for 24 hours. Osmolarity, vitamin A, fibronectin, platelet-derived growth factor-AB, vascular endothelial growth factor, epithelial growth factor, and transforming growth factor-β1 were quantified. The proliferative and migratory potential of the eye drops was assayed on primary human keratocytes.
Platelet-derived growth factor-AB, vascular endothelial growth factor, epithelial growth factor, and vitamin A levels remained constant for each time and for each storage condition, whereas fibronectin, transforming growth factor-β1, and osmolarity values were slightly modified after freezing. Cell proliferation and migration were significantly enhanced with the biological eye drops independently of the time and the storage condition. No microbial contamination was observed in any plasma rich in growth factors eye drops.
Plasma rich in growth factors eye drops can be stored for up to 3 months without any reduction of the main proteins involved in ocular surface healing. Their use during 24 hours either at 4°C or at RT did not alter the composition and the in vitro biological activity of the eye drops.
我们评估了富含生长因子的滴眼液在-20°C储存3个月后以及在4°C或室温(RT)储存24小时后,与0时刻获得的样本(新鲜样本)相比,其成分和生物活性是否保持不变。
采集10名健康供体的血液,离心,并通过避免收集白膜制备富含生长因子的血浆。滴眼液保持新鲜或在-20°C储存15、30和90天。每次,将2份等分试样在室温或4°C下保存24小时。对渗透压、维生素A、纤连蛋白、血小板衍生生长因子AB、血管内皮生长因子、上皮生长因子和转化生长因子-β1进行定量。在原代人角膜细胞上测定滴眼液的增殖和迁移潜力。
血小板衍生生长因子AB、血管内皮生长因子、上皮生长因子和维生素A水平在每次和每种储存条件下均保持恒定,而纤连蛋白、转化生长因子-β1和渗透压值在冷冻后略有改变。生物滴眼液可显著增强细胞增殖和迁移,且与时间和储存条件无关。在任何富含生长因子的血浆滴眼液中均未观察到微生物污染。
富含生长因子的滴眼液可储存长达3个月,而不会降低参与眼表愈合的主要蛋白质的含量。在4°C或室温下24小时内使用不会改变滴眼液的成分和体外生物活性。