Custo Scott, Baron Byron, Felice Alex, Seria Elisa
Department of Physiology & Biochemistry, Faculty of Medicine & Surgery, University of Malta, Msida, Malta.
Centre for Molecular Medicine & Biobanking, University of Malta, Msida, Malta.
GMS Interdiscip Plast Reconstr Surg DGPW. 2022 Jul 5;11:Doc06. doi: 10.3205/iprs000167. eCollection 2022.
Platelet-derived products have been shown as promising novel therapeutic agents for chronic wounds. However, their clinical use requires a greater degree of method standardisation, part of which involved more extensive cataloguing of their biochemical composition. This study aimed to quantify and compare total protein and 6 angiogenically-active growth factors in three distinct platelet products.
Platelet Lysate (PL, n=5), Calcium-activated Platelet Rich Plasma (Ca-PRP, n=5) and Platelet-Rich Fibrin (PRF, n=5) were prepared from pooled platelet apheresis products (n=10). Ca-PRP and PRF were prepared from the same units (n=5) by activation with 20 mmolL calcium chloride. PL was prepared from the remaining (n=5) units using an established lysate. Total protein was quantified with the Bradford Assay. Sandwich enzyme-linked immunosorbent assay was used to quantify six growth factors: epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), stromal cell derived growth factor-1α (SDF-1α), endostatin, and transforming growth factor-β1 (TGF-β1).
Protein retrieval differed significantly (p<0.05) between the three products: PL (11.35±0.80 mg/mL) < Ca-PRP (20.44±8.17 mg/mL) < PRF (40.67±3.13 mg/mL). Growth factor yield was considerable in all three products and differed significantly for: VEGF (PL<PRF); EGF (Ca-PRP<PRF); HFG (PL<Ca-PRP); Endostatin (PL<Ca-PRP, PRF<Ca-PRP, PL<PRF) and TGF-β1 (Ca-PRP<PL, Ca-PRP<PRF).
Platelet apheresis products contain a substantial quantity of the investigated pro- and anti-angiogenic growth factors. Their release varies depending on the manufacturing protocol used. Clinically, alternate products could thus be combined to provide a therapeutically optimal mix of growth factors.
血小板衍生产品已被证明是治疗慢性伤口的有前景的新型治疗剂。然而,它们的临床应用需要更高程度的方法标准化,其中一部分涉及对其生化组成进行更广泛的编目。本研究旨在量化和比较三种不同血小板产品中的总蛋白和6种血管生成活性生长因子。
从混合的血小板单采产品(n = 10)中制备血小板裂解物(PL,n = 5)、钙激活富血小板血浆(Ca-PRP,n = 5)和富血小板纤维蛋白(PRF,n = 5)。Ca-PRP和PRF由相同的单位(n = 5)通过用20 mmol/L氯化钙激活制备。PL由其余(n = 5)单位使用既定的裂解物制备。用Bradford法对总蛋白进行定量。采用夹心酶联免疫吸附测定法对六种生长因子进行定量:表皮生长因子(EGF)、血管内皮生长因子(VEGF)、肝细胞生长因子(HGF)、基质细胞衍生生长因子-1α(SDF-1α)、内皮抑素和转化生长因子-β1(TGF-β1)。
三种产品之间的蛋白质回收率差异显著(p<0.05):PL(11.35±0.80 mg/mL)<Ca-PRP(20.44±8.17 mg/mL)<PRF(40.67±3.13 mg/mL)。所有三种产品中的生长因子产量都相当可观,并且在以下方面差异显著:VEGF(PL<PRF);EGF(Ca-PRP<PRF);HFG(PL<Ca-PRP);内皮抑素(PL<Ca-PRP, PRF<Ca-PRP, PL<PRF)和TGF-β1(Ca-PRP<PL, Ca-PRP<PRF)。
血小板单采产品含有大量研究的促血管生成和抗血管生成生长因子。它们的释放因所使用的制造方案而异。因此,在临床上,可以将替代产品组合起来,以提供治疗上最佳的生长因子组合。