The effect of short-term cryopreservation on the properties and functionality of platelet-rich plasma

Platelets. 2023 Dec;34(1):2210243. doi: 10.1080/09537104.2023.2210243.

Abstract

Platelet Rich Plasma (PRP) is a biological treatment which, thanks to its enhanced growth factors content, is widely used in the field of regenerative medicine for its reparative effects. Although it is usually used fresh immediately after preparation, its freezing for preservation for future usage could be key in increasing its versatility and new applications. To assess the suitability of freezing, after collecting PRP and platelet lysates (PL) from 6 patients, they were preserved for 1 or 3 months at temperatures of -20ºC and -80°C. Measurements were then made on platelet number and integrity, growth factor levels, biomechanical properties of the clot and its bioactivity on cultured cells. Fresh PRP and PL were used as controls. The results showed an increase in platelet size (p < .01) and clot elasticity (p < .01), as well as decrease in levels of PDGF (P < .05) and VEGF (p < .05), though the overall bioactivity was not affected as culture cells showed the same responsiveness to both frozen and fresh PRP and PL in terms of cell viability. Based on these results, it could be assumed that preservation of PRP by freezing is a feasible and suitable option for its further use.

Keywords: Bioactivity; cryopreservation; growth factors; platelet-rich plasma; storage.

Plain language summary

What is the context? Platelet-Rich Plasma (PRP) is a biological treatment widely used in regenerative medicine as a result of its high content of growth factors.In its routine use, PRP has autologous character, since it is obtained from the same patient and its infiltration in the affected area takes place immediately after it is obtained.Its storability would give PRP versatility in use, which could enable a potential allogeneic use and even significantly reduce the number of blood draws for each patient.It is necessary to establish a PRP storage protocol where its properties are not affected.What is new? PRP was stored at two time points (1 and 3 months) and temperatures (−20ºC and −80ºC) in activated and unactivated states. Afterwards, platelet number, size and activation were measured. Furthermore, the biomechanical properties of the resulting clot, the growth factor content and PRP’s impact on cell viability were analyzed.The limiting factor was not having used aggregometry or other techniques that measure other cellular processes, as well as the limited sample size.The results showed that freezing affected platelet size, the levels of platelet-derived GFs and the biomechanical properties of the clot. However, plasmatic levels of growth factors or its capacity to boost cellular proliferation were not affected.What is the impact?The clinical impact of this work is the ability to preserve PRP by freezing. This is especially relevant as it allows a possible use of PRP as an allogeneic treatment. Moreover, its preservation significantly reduces the number of blood draws for each patient, especially in those with puncture difficulties or with apprehension.

MeSH terms

  • Cell Culture Techniques
  • Cells, Cultured
  • Cryopreservation*
  • Humans
  • Intercellular Signaling Peptides and Proteins / metabolism
  • Platelet-Rich Plasma* / metabolism

Substances

  • Intercellular Signaling Peptides and Proteins