Karsmarski Owen P, Hawthorne Benjamin C, Cusano Antonio, LeVasseur Matthew R, Wellington Ian J, McCarthy Mary Beth, Cote Mark P, Mazzocca Augustus D
Department of Orthopaedic Surgery, University of Connecticut, Farmington, CT 06032, USA.
J Clin Med. 2022 Jun 15;11(12):3442. doi: 10.3390/jcm11123442.
The purpose of this study was to investigate proteomic alteration that occurs to whole blood when converted to activated serum (AS) using an autologous thrombin system. This study further sought to evaluate the functional in vitro effect of AS on tenocytes, chondrocytes, subacromial bursal cells, and osteoblasts. The peptide/protein composition of AS was analyzed by liquid chromatography−mass spectrophotometry (LC-MS). The cell lines were treated with AS, and cellular proliferation was quantified 48 h after treatment. Platelet-derived growth factor (PDGF), insulin-like growth factor 1 (IGF-1), vascular endothelial growth factor (VEGF), interleukin-1 beta (IL-1β), and interleukin-1 receptor antagonist (IL-1Ra) were quantified utilizing enzyme-linked immunosorbent assays (ELISAs). LC-MS identified 357 proteins across the AS and whole blood. Fifty-four of the proteins identified had significant differences between the relative protein abundance of the AS samples compared to whole blood. Treatment with AS in all cell lines significantly increased proliferation compared to control cells at 48 h. Increased PDGF, VEGF, and IGF-1 in all cell lines exposed to AS compared to the control (p < 0.05) were observed. These findings suggest that treatment with AS increases in vitro cellular proliferation and the release of growth factors that may play a role in tissue repair.
本研究的目的是调查使用自体凝血酶系统将全血转化为活化血清(AS)时全血中发生的蛋白质组学变化。本研究进一步试图评估AS对肌腱细胞、软骨细胞、肩峰下滑囊细胞和成骨细胞的体外功能作用。通过液相色谱 - 质谱法(LC-MS)分析AS的肽/蛋白质组成。用AS处理细胞系,并在处理后48小时对细胞增殖进行定量。利用酶联免疫吸附测定(ELISA)对血小板衍生生长因子(PDGF)、胰岛素样生长因子1(IGF-1)、血管内皮生长因子(VEGF)、白细胞介素-1β(IL-1β)和白细胞介素-1受体拮抗剂(IL-1Ra)进行定量。LC-MS在AS和全血中鉴定出357种蛋白质。与全血相比,所鉴定的54种蛋白质在AS样品的相对蛋白质丰度之间存在显著差异。与对照细胞相比,在48小时时用AS处理所有细胞系均显著增加了增殖。与对照相比,在所有暴露于AS的细胞系中观察到PDGF、VEGF和IGF-1增加(p < 0.05)。这些发现表明,用AS处理可增加体外细胞增殖以及可能在组织修复中起作用的生长因子的释放。