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Biological Stability of Plasma Rich in Growth Factors Eye Drops After Storage of 3 Months

Anitua, Eduardo MD, DDS; Muruzabal, Francisco PhD; Pino, Ander MSc; Merayo-Lloves, Jesús MD, PhD; Orive, Gorka PhD

doi: 10.1097/ICO.0b013e31829f7088
Basic Investigation

Purpose: 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).

Methods: 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.

Results: 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.

Conclusions: 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.

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*Fundación Eduardo Anitua, Vitoria, Spain; and

Fundación de Investigación Oftalmológica, Instituto Oftalmológico Fernández-Vega, Oviedo, Spain.

Reprints: Eduardo Anitua, Instituto Eduardo Anitua, c/Jose Maria Cagigal 19, 01005 Vitoria, Spain (e-mail:

Supported by the Customized Eye Care, CEYEC (No. CEN-20091021) project, which has been supported by the Centre for Industrial Technological Development in the fifth edition of the CENIT program. The aim of this program is to promote the public–private stable cooperation in research, development, and innovation, which is part of the Spanish government initiative INGENIO 2010.

E. Anitua, F. Muruzabal, A. Pino, and G. Orive are scientists at Foundation Eduardo Anitua, which investigates the potential of plasma rich in growth factors in regenerative medicine.

Supplemental digital content is available for this article. Direct URL citations appear in the printed text and are provided in the HTML and PDF versions of this article on the journal’s Web site (

Received April 12, 2013

Accepted June 06, 2013

Copyright © 2013 Wolters Kluwer Health, Inc. All rights reserved.