Wachtel Nikolaus, Weber Luisa, Moellhoff Nicholas, Kuhlmann Constanze, Giunta Riccardo E, Alberton Paolo, Ehrl Denis, Wiggenhauser Severin
Division of Hand, Plastic and Aesthetic Surgery, University Hospital, LMU Munich, Germany.
Department of Plastic, Reconstructive and Hand Surgery, Burn Center for Severe Burn Injuries, Klinikum Nuremberg Hospital, Paracelsus Medical University, Germany.
Wound Repair Regen. 2025 May-Jun;33(3):e70040. doi: 10.1111/wrr.70040.
Recent studies showcased the regenerative potential of Platelet-Rich Fibrin (PRF) combined with Adipose-Derived Stem Cells (ASC). PRF enhances cellular proliferation through sustained growth factor secretion which are continuously released to surrounding cells. However, its regulatory mechanisms remain unclear. ASC were isolated from liposuction and abdominoplasty samples of healthy donors, characterised via flow-cytometry and cultured for 7 days. Four cell culture conditions were tested: (1) 10% PRF extract (PRFe), (2) 10% Platelet-Low Plasma (PLP), (3) 10% Foetal Bovine Serum (FBS) and (4) basal medium as control. Cell viability and proliferation were assessed using AlamarBlue and PicoGreen assays, as well as live-dead staining. Enzyme-Linked Immunosorbent Assays quantified growth factor concentrations, while multiplex qPCR and immunocytochemical staining analysed gene and protein expression on days 1 and 7. PRFe-supplemented cultures showed the highest viability and proliferation, significantly surpassing other groups at day 7 (p < 0.05). Supernatant analysis revealed significantly elevated TGF-β1 and PDGF-AA/BB levels in PRFe cultures at day 7 (p of at least < 0.05). Multiplex qPCR indicated increased expression of proliferation and pluripotency markers (NANOG, JUN, SOX2, RPS6KA4; p < 0.05) and fibrillar collagen (COL1A; p < 0.05) in the PRFe group. These findings demonstrate that PRFe significantly enhances ASC proliferation and regenerative potential. Elevated levels of TGF-1, PDGF-AA/BB and to a lesser extend VEGF in PRFe cultures suggest that its benefits in regenerative medicine may be linked to these cytokines' upregulation. These results underscore PRFe's potential as a key supplement for optimising ASC-based therapies in tissue regeneration.
近期研究展示了富含血小板纤维蛋白（PRF）与脂肪来源干细胞（ASC）联合应用的再生潜力。PRF通过持续分泌生长因子来增强细胞增殖，这些生长因子会持续释放到周围细胞中。然而，其调控机制仍不清楚。从健康供体的抽脂和腹部整形样本中分离出ASC，通过流式细胞术进行表征，并培养7天。测试了四种细胞培养条件：（1）10% PRF提取物（PRFe），（2）10%低血小板血浆（PLP），（3）10%胎牛血清（FBS），（4）基础培养基作为对照。使用AlamarBlue和PicoGreen检测法以及活死染色评估细胞活力和增殖。酶联免疫吸附测定法定量生长因子浓度，而多重qPCR和免疫细胞化学染色分析第1天和第7天的基因和蛋白质表达。补充PRFe的培养物显示出最高的活力和增殖，在第7天显著超过其他组（p < 0.05）。上清液分析显示，第7天时PRFe培养物中转化生长因子-β1（TGF-β1）和血小板衍生生长因子-AA/BB（PDGF-AA/BB）水平显著升高（p至少< 0.05）。多重qPCR表明PRFe组中增殖和多能性标志物（NANOG、JUN、SOX2、RPS6KA4；p < 0.05）以及纤维状胶原蛋白（COL1A；p < 0.05）的表达增加。这些发现表明PRFe显著增强了ASC的增殖和再生潜力。PRFe培养物中TGF-1、PDGF-AA/BB水平升高以及程度较轻的血管内皮生长因子（VEGF）升高表明，其在再生医学中的益处可能与这些细胞因子的上调有关。这些结果强调了PRFe作为优化基于ASC的组织再生疗法的关键补充剂的潜力。
