Wahyuningsih Komang Ardi, Sobariah Siti, Rosliana Iis, Rosadi Imam, Widyastuti Tias, Afini Irsyah, Wanandi Septelia Inawati, Soewondo Pradana, Wibowo Heri, Pawitan Jeanne Adiwinata
Doctoral Program of Biomedical Sciences, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia.
Hayandra Lab, Yayasan Hayandra Peduli, Jakarta, Indonesia.
Stem Cell Investig. 2019 Dec 20;6:43. doi: 10.21037/sci.2019.10.02. eCollection 2019.
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.
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 °C, 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.
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 °C 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.
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.
富含血小板血浆(PRP)含有促血管生成生长因子,包括血管内皮生长因子(VEGF)。血管生成是糖尿病足溃疡(DFU)伤口愈合的必要组成部分。PRP的成分因方法和供体健康状况而异。我们的团队已经开发出一种改进的用于糖尿病治疗的PRP方案。本研究的目的是检测这些糖尿病患者和非糖尿病患者群体中促血管生成因子VEGF的水平。
使用来自Klinik Hayandra登记的13名糖尿病患者和10名非糖尿病患者的24 mL全血制备PRP。将枸橼酸钠管中的全血以1000 rpm离心5分钟,然后进行血浆分离。血浆样本以3000 rpm离心5分钟。弃去上层贫血小板血浆层,留下5 mL浓缩的含血小板血浆(PRP)。将浓缩血浆样本混合、分装,储存在-86°C,并合并用于血小板计数、VEGF和总蛋白分析。还使用新鲜全血和PRP进行血小板计数,以测量PRP制备后的变化。
糖尿病供体的全血血小板计数高于非糖尿病供体,但这种差异无统计学意义。两组使用我们的方法制备PRP后,血小板数量平均增加超过250%。无论糖尿病状态如何,在-86°C冻融样本会使90%以上的PRP血小板裂解。糖尿病PRP的平均总蛋白含量较低,VEGF浓度较高。糖尿病供体裂解的血小板释放的VEGF比非糖尿病供体的更多。
糖尿病供体的PRP具有更高的VEGF含量,使得自体PRP应用成为DFU的一种有前景的治疗方法。然而,这应该在另一项合适的临床试验中进行研究。