An optimised protocol for platelet-rich plasma preparation to improve its angiogenic and regenerative properties

Abstract

Although platelet-rich plasma (PRP) is used as a source of growth factors in regenerative medicine, its effectiveness remains controversial, partially due to the absence of PRP preparation protocols based on the regenerative role of platelets. Here, we aimed to optimise the protocol by analysing PRP angiogenic and regenerative properties. Three optimising strategies were evaluated: dilution, 4 °C pre-incubation, and plasma cryoprecipitate supplementation. Following coagulation, PRP releasates (PRPr) were used to induce angiogenesis in vitro (HMEC-1 proliferation, migration, and tubule formation) and in vivo (chorioallantoic membrane), as well as regeneration of excisional wounds on mouse skin. Washed platelet releasates induced greater angiogenesis than PRPr due to the anti-angiogenic effect of plasma, which was decreased by diluting PRPr with saline. Angiogenesis was also improved by both PRP pre-incubation at 4 °C and cryoprecipitate supplementation. A combination of optimising variables exerted an additive effect, thereby increasing the angiogenic activity of PRPr from healthy donors and diabetic patients. Optimised PRPr induced faster and more efficient mouse skin wound repair compared to that induced by non-optimised PRPr. Acetylsalicylic acid inhibited angiogenesis and tissue regeneration mediated by PRPr; this inhibition was reversed following optimisation. Our findings indicate that PRP pre-incubation at 4 °C, PRPr dilution, and cryoprecipitate supplementation improve the angiogenic and regenerative properties of PRP compared to the obtained by current methods.

Platelet‐rich plasma counteracts detrimental effect of high‐glucose concentrations on mesenchymal stem cells from Bichat fat pad

“…However, as compared with nondiabetic controls, angiogenic activities of platelets remain unaltered in T2D patients (Miao, Zhang, Huang, & Li, 2016). Some authors have also found that PRP from diabetic patients releases more VEGF and increases proliferation and migration of endothelial cells at larger extent, compared with PRP from healthy donors (Etulain et al, 2018; Karina et al, 2019). Notably, solid evidence supports the use of autologous PRP for the treatment of diabetic foot ulcers (Hirase, Ruff, Surani, & Ratnani, 2018).…”

Functionalising Collagen-Based Scaffolds With Platelet-Rich Plasma for Enhanced Skin Wound Healing Potential

“…The effects of the released factors were evaluated for both our composite scaffold and PRP releasate (PRPr) in different cell types (endothelial, mesenchymal stromal cells, macrophages, fibroblasts, and keratinocytes) since all of them have key roles during different stages of wound healing (Gurtner et al, 2008;Eming et al, 2014). There is vast literature suggesting that PRP supports endothelial cell proliferation and induces angiogenesis/vascularization both in vitro and in vivo (Kakudo et al, 2014;Anitua et al, 2015;Notodihardjo et al, 2015;Etulain et al, 2018;Romaldini et al, 2019;Samberg et al, 2019). Our work corroborates those findings, but, importantly, shows that this activity is maintained within a biomaterial scaffold with proven regenerative potential.…”

Intravesical injections of platelet‐rich plasma is effective and safe in treatment of interstitial cystitis refractory to conventional treatment—A prospective clinical trial

“…However, the temperature, the leukocyte, and the plasma volume might influence therapeutic results have not been well determined. Recent investigation showed that adding saline instead of plasma (which contains anti‐platelet factors) to the platelet pellets could improve the therapeutic effects on would healing and angiogenesis in animal model . In this study, a volume of 10 mL PRP was used simply because we can inject PRP at 20 sites through most of the bladder wall.…”