Evaluation of platelet-rich plasma from diabetic donors shows increased platelet vascular endothelial growth factor release

Abstract

Background: Platelet-rich plasma (PRP) contains pro-angiogenic growth factors including vascular endothelial growth factor (VEGF). Angiogenesis is a necessary component of wound healing in instances of diabetic foot ulcers (DFU). PRP composition varies depending on methods and donor health status. Our group has developed an improved PRP protocol for diabetes treatment. The aims of this study were to examine the levels of the pro-angiogenic factor VEGF in these patient populations with and without diabetes.Methods: PRP was prepared using 24 mL of whole blood from 13 diabetic and 10 non-diabetic patients registered at Klinik Hayandra. Whole blood in sodium citrate tubes were centrifuged at 1,000 rpm for 5 minutes followed by plasma separation. Plasma samples were centrifuged at 3,000 rpm for 5 minutes.Upper platelet-poor plasma layers were discarded, leaving 5 mL of concentrated platelet containing plasma (PRP). Concentrated plasma samples were mixed, aliquoted, stored at −86 ℃, and pooled for platelet count, VEGF, and total protein analyses. Platelet counting was also performed using fresh whole blood and PRP to measure changes following PRP preparation.Results: Diabetic donors had higher whole blood platelet counts than non-diabetic donors, but this difference was not statistically significant. An average increase of more than 250% in platelet number after PRP preparation using our method was noted in both groups. Freezing-thawing samples at −86 ℃ lysed more than 90% of PRP platelets regardless of diabetes status. Diabetic PRP had lower mean total protein and higher VEGF concentrations. Lysed platelets from diabetic donors released more VEGF than those from non-diabetic donors.Conclusions: PRP from diabetic donors had higher VEGF content making autologous PRP application a promising treatment for DFU. However, this should be investigated another appropriate clinical trial.

Safety of Technique and Procedure of Stromal Vascular Fraction Therapy: From Liposuction to Cell Administration

“…Marx reported that double centrifugation of autologous blood can produce PRP with concentration of platelets of at least 1 × 10 6 cell/ µ L that is almost 5 times higher than the platelet concentration in the whole blood [ 45 ]. Platelets from majority of our patients, especially those who have diabetes, were easier to aggregate after second centrifugation as we observed in laboratory, which may reflect their behavior in blood circulation [ 46 ]. Aggregated platelet releases various cytokines and growth factors that in turn activate endothelium [ 47 ].…”

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).…”

The application of platelet‐rich plasma for skin graft enrichment: A meta‐analysis

“…For example, VEGF in PRP mainly takes part in angiogenesis. 31 EGF is a mitogenic and chemotactic factor for fibroblasts and epithelial cells. 32 PDGF is also a mitogenic and chemotactic protein for fibroblasts and endothelial progenitor cells.…”

“…Through launching signalling cascades, these growth factors will initiate multiple cellular biochemical changes. For example, VEGF in PRP mainly takes part in angiogenesis 31 . EGF is a mitogenic and chemotactic factor for fibroblasts and epithelial cells 32 .…”