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富含生长因子眼药水的短期和长期稳定性

Short- and Long-Term Stability of Plasma Rich in Growth Factors Eye Drops.

作者信息

Anitua Eduardo, de la Fuente María, Muruzábal Francisco, Merayo-Lloves Jesús

机构信息

BTI Biotechnology Institute, Vitoria, Spain.

University Institute for Regenerative Medicine and Oral Implantology-UIRMI (UPV/EHU-Fundación Eduardo Anitua), Vitoria, Spain.

出版信息

Cornea. 2021 Jan;40(1):107-112. doi: 10.1097/ICO.0000000000002519.

Abstract

PURPOSE

To analyze whether plasma rich in growth factors (PRGFs) eye drops preserve their activity and biological properties after storage for 9 and 12 months at -20°C, and at 4°C, and at room temperature (RT) for 3 and 7 days in comparison to fresh samples (t0).

METHODS

PRGF eye drops were obtained from 6 healthy donors. Then, they were stored for 9 and 12 months at -20°C. At each time, different PRGF eye drops samples were thawed and maintained at RT or at 4°C for 3 and 7 days. Platelet-derived growth factor-AB, epidermal growth factor, transforming growth factor-β1, vascular endothelial growth factor, angiopoietin-1, and thrombospondin-1 were analyzed at each time and temperature of storage. In addition, the pH level, the microbial contamination, and the proliferative potential on primary human corneal stromal fibroblasts human keratocytes of each obtained PRGF eye drops were also evaluated.

RESULTS

All growth factor levels were preserved at each time and storage condition. No differences were observed on the human keratocytes proliferation after treatment with PRGF eye drops at any studied time or temperature. No microbial contamination was observed in any of the PRGF eye drops. Finally, the pH levels increased significantly after 9 and 12 months of storage at -20°C compared with t0.

CONCLUSIONS

PRGF eye drops can be stored for up to 12 months without reduction of the main growth factors and proteins and without any microbial contamination. Furthermore, the biological activity of the PRGF eye drops is maintained after storing for 3 and 7 days at 4°C or at RT.

摘要

目的

分析富含生长因子的血浆(PRGF)滴眼液在-20°C下储存9个月和12个月、在4°C下储存以及在室温(RT)下储存3天和7天后,与新鲜样本(t0)相比,是否仍保留其活性和生物学特性。

方法

从6名健康供体获取PRGF滴眼液。然后,将其在-20°C下储存9个月和12个月。每次,将不同的PRGF滴眼液样本解冻,并在室温或4°C下保持3天和7天。在每次储存时间和温度下分析血小板衍生生长因子-AB、表皮生长因子、转化生长因子-β1、血管内皮生长因子、血管生成素-1和血小板反应蛋白-1。此外,还评估了每种获得的PRGF滴眼液的pH值水平、微生物污染情况以及对原代人角膜基质成纤维细胞和人角膜细胞的增殖潜力。

结果

在每次储存时间和条件下,所有生长因子水平均得以保留。在任何研究的时间或温度下,用PRGF滴眼液处理后人角膜细胞增殖均未观察到差异。在任何PRGF滴眼液中均未观察到微生物污染。最后,与t0相比,在-20°C下储存9个月和12个月后pH值水平显著升高。

结论

PRGF滴眼液可储存长达12个月,主要生长因子和蛋白质不会减少,也不会有任何微生物污染。此外,PRGF滴眼液在4°C或室温下储存3天和7天后仍保持其生物活性。

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