Kühnel Harald, Pasztorek Markus, Kuten-Pella Olga, Kramer Karina, Bauer Christoph, Lacza Zsombor, Nehrer Stefan
Center for Regenerative Medicine, University for Continuing Education Krems, Dr.-Karl-Dorrek-Straße 30, 3500 Krems an der Donau, Austria.
Department of Applied Life Science, Bioengineering, FH-Campus Vienna, Favoritenstrasse 222, 1100 Vienna, Austria.
Curr Issues Mol Biol. 2024 Feb 28;46(3):1865-1885. doi: 10.3390/cimb46030122.
Blood-derived products, such as citrate platelet-rich plasma (CPRP) and hyperacute serum (HAS), are recognized for their rich growth factor content. When human dermal fibroblast (HDF) cells are exposed to combined mitogenic and DNA-damaging stimuli, it can lead to an increased burden of senescent cells and a modified senescence-associated secretory phenotype. In this study, the senescent state was comprehensively assessed through various methods, including phosphorylated histone H2AX (γH2AX) staining, p21 and p16 q-PCR, p21-western blot, growth curves, and senescence-associated ß-galactosidase staining. Two primary treatments with blood products were administered, one early (immediately after etoposide) and the other late (11 days after etoposide treatment). The effects of the blood product treatment were evaluated by measuring interleukin 6 and 8 (IL-6 and IL-8) levels, as well as collagen 1 (COL1) and p21 mRNA expression. Additionally, 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assays, cell size measurements, viability assays, and cell number calculations were conducted. The results revealed that cells treated with hyperacute serum in the early treatment phase exhibited the lowest observed IL-6 and IL-8 levels. In contrast, a clear inflammatory response for IL-8 was observed in cells treated with hyperacute serum and citrate platelet-rich plasma during the late treatment. Furthermore, an upregulation of COL1 expression was observed in the early treatment, while cells in the late treatment group remained unaffected. Notably, citrate platelet-rich plasma-treated cells showed a decrease in COL1 expression. Overall, the treatment with blood products appears to have slightly positive effects on skin rejuvenation.
血液衍生产品,如枸橼酸富血小板血浆(CPRP)和超急性血清(HAS),因其富含生长因子而闻名。当人皮肤成纤维细胞(HDF)暴露于有丝分裂原和DNA损伤联合刺激时,会导致衰老细胞负担增加和衰老相关分泌表型改变。在本研究中,通过多种方法全面评估衰老状态,包括磷酸化组蛋白H2AX(γH2AX)染色、p21和p16定量聚合酶链反应(q-PCR)、p21蛋白免疫印迹、生长曲线以及衰老相关β-半乳糖苷酶染色。进行了两种血液制品的主要处理,一种是早期处理(依托泊苷后立即处理),另一种是晚期处理(依托泊苷处理后11天)。通过测量白细胞介素6和8(IL-6和IL-8)水平以及1型胶原蛋白(COL1)和p21信使核糖核酸(mRNA)表达来评估血液制品处理的效果。此外,还进行了2,3-双-(2-甲氧基-4-硝基-5-磺基苯基)-2H-四唑-5-甲酰苯胺(XTT)检测、细胞大小测量、活力检测和细胞数量计算。结果显示,在早期处理阶段用超急性血清处理的细胞中,观察到的IL-6和IL-8水平最低。相比之下,在晚期处理期间,用超急性血清和枸橼酸富血小板血浆处理的细胞中观察到明显的IL-8炎症反应。此外,在早期处理中观察到COL1表达上调,而晚期处理组的细胞未受影响。值得注意的是,枸橼酸富血小板血浆处理的细胞显示COL1表达下降。总体而言,血液制品处理似乎对皮肤年轻化有轻微的积极作用。